Patent Application
20250177281
The present invention relates to a composition, in particular a cosmetic composition, comprising a) at least one polyhydroxyalkanoate (PHA) copolymer bearing (un)saturated hydrocarbon-based groups, b) at least one oligo/polyester, c) optionally at least one fatty substance, and d) optionally at least one organic solvent other than c), and also to a process for treating keratin materials using such a composition.
It is known practice to use, in cosmetics, film-forming polymers which can be conveyed in organic media, such as hydrocarbon-based oils. Polymers are notably used as film-forming agents in makeup products such as mascaras, eyeliners, eyeshadows or lipsticks.
FR-A-2964663 describes a cosmetic composition comprising pigments coated with a C3-C21 polyhydroxyalkanoate, such as poly(hydroxybutyrate-co-hydroxyvalerate).
WO 2011/154508 describes a cosmetic composition comprising a 4-carboxy-2-pyrrolidinone derivative and a film-forming polymer which may be a polyhydroxyalkanoate, such as polyhydroxybutyrate, polyhydroxyvalerate and polyhydroxybutyrate-co-polyhydroxyvalerate.
US-A-2015/274972 describes a cosmetic composition comprising a thermoplastic resin, such as a polyhydroxyalkanoate, in aqueous dispersion and a silicone elastomer.
The majority of the polyhydroxyalkanoate copolymers are polymers derived from the polycondensation of polymeric repeating units that are for the most part identical and derived from the same carbon source or substrate. These documents do not describe the cosmetic use of copolymers derived from polycondensation using an aliphatic substrate or first carbon source, and at least one second substrate different from the first, comprising one or more (un)saturated hydrocarbon-based groups with oligo/polyesters. There is thus a need for a composition comprising polyhydroxyalkanoate copolymers which are lipophilic or soluble in a fatty phase. This makes it possible to obtain a film on keratin materials which has good cosmetic properties, notably good resistance to oils and to sebum, and also to be able to modify the gloss or the mattness.
The Applicant has discovered that polyhydroxyalkanoate copolymers bearing particular grafted or functionalized hydrocarbon-based groups, as defined below, may be readily used in fatty media, thus making it possible to obtain homogeneous compositions. Composition C1 shows good stability, notably after storage for one month at room temperature (25° C.). Composition C1, notably after its application to keratin materials, makes it possible to obtain a film having good cosmetic properties, good persistence of the colour without running, and also a matt or glossy appearance of the treated keratin materials.
Patent application EP 2 699 636 discloses makeup compositions rich in monoalcohol and comprising an oligo/polyester chosen from vinyl polymers grafted with a carbosiloxane dendrimer in order to obtain better persistence of the matt effect. Alcoholic makeup compositions based on red organic pigments are also known in patent FR 3 005 857. These documents do not mention the problems of resistance to rubbing and do not use PHAs. In addition, these compositions, which contain oils, are not entirely satisfactory in terms of comfort on application.
In many conditions of use of film-forming materials on keratin materials, for instance in makeup or colouring applications, it is desirable to have, in addition to good resistance to water and oils, notably food oils such as olive oil, very good resistance to rubbing of the deposits of film-forming materials both to avoid transfer, for example onto clothing, and to maintain a homogeneous appearance of the deposits. If the resistance to rubbing is insufficient, the deposits obtained can quickly become very unsightly for consumers, in particular if these deposits are coloured as in makeup applications such as lipsticks, foundations or mascaras. In hair applications, the absence of resistance to rubbing is also very problematic in all colouring applications since it gives rise to transfer onto clothing and creates an unsightly appearance of the keratin fibres. There is thus a need to improve the persistence of PHAs conveyed in an aqueous phase.
There is thus a real need to obtain deposits of film-forming materials that are resistant to oils, notably food oils, and that are water-resistant and have very good resistance to rubbing.
When the deposit is coloured, these problems of resistance to humidity lead to a transfer of colour, for example onto clothing, which is in itself problematic and which makes the deposit very unsightly. It is thus important for the deposit to be also water-resistant.
These problems are solved by the use of the compositions C1 described hereinbelow, these compositions making it possible to significantly improve the resistance to rubbing of polyhydroxyalkanoate (PHA) copolymer(s). Furthermore, the compositions C1 according to the invention make it possible to obtain, after deposition, a film on keratin materials which has good cosmetic properties, notably good resistance to oils and to sebum, and good water resistance, and also to be able to modify the gloss or the mattness.
Thus, the main subject of the present invention is a composition C1, in particular a cosmetic composition, comprising:
-[—O—CH(R1)—CH2—C(O)—]- unit (A)
-[—O—CH(R2)—CH2—C(O)—]- unit (B)
Preferably, composition C1 contains c) and more preferentially the ingredients c) and d).
Another object of the invention is the cosmetic use of a composition C1 comprising a) one or more PHA copolymers as defined previously, b) one or more oligo/polyesters as defined previously, optionally c) one or more fatty substances as defined previously, d) optionally one or more organic solvents other than c), and e) optionally water; preferably, composition C1 contains the ingredients c)+d).
Another subject of the invention is a process for treating keratin materials, preferably a) keratin fibres, notably human keratin fibres such as the hair, or p) human skin, in particular the lips, using a) one or more PHA copolymers as defined previously, b) one or more oligo/polyesters, optionally c) one or more fatty substances as defined previously, optionally d) one or more organic solvents other than c) and optionally e) water; preferably, composition C1 contains the ingredients c)+d).
More particularly, a subject of the invention is a non-therapeutic cosmetic process for treating keratin materials, comprising the application to the keratin materials of a composition C1 as defined previously. The treatment process is in particular a process for caring for or making up keratin materials.
For the purposes of the present invention and unless otherwise indicated:
Furthermore, unless otherwise indicated, the limits delimiting the extent of a range of values are included in that range of values.
Composition C1 of the invention comprises as first ingredient a) one or more PHA copolymers which contain, or preferably consist of, at least two different repeating polymer units chosen from the units (A) as defined previously and (B) as defined below.
Composition C1 of the invention, preferably a cosmetic composition, comprises:
-[—O—CH(R1)—CH2—C(O)—]- unit (A)
-[—O—CH(R2)—CH2—C(O)—]- unit (B)
According to one embodiment, composition C1 contains the ingredients c) and d).
The term “co-polymer” means that said polymer is derived from the polycondensation of polymeric repeating units that are different from each other, i.e. said polymer is derived from the polycondensation of polymeric repeating units (A) that are different from each other, or from the polycondensation of polymeric repeating units (A) with (B), it being understood that the polymeric units (A) are different from the polymeric units (B), it being possible for said copolymer to be obtained from a single saturated or unsaturated aliphatic carbon source which is optionally substituted and/or interrupted, preferably unsubstituted and uninterrupted, or from several carbon sources, in particular at least one of which is an uninterrupted unsubstituted saturated aliphatic and the other carbon source(s) are saturated or unsaturated aliphatic, optionally substituted notably with a halogen atom such as bromine, or with a cyano group, a Bunte salt, a dithiolane radical, a carboxyl, etc.
According to one embodiment, the copolymer according to the invention is derived from a single carbon source, preferably a single saturated or unsaturated aliphatic carbon source which is optionally substituted and/or interrupted, preferably unsubstituted and uninterrupted.
According to one embodiment, the copolymer according to the invention is derived from several carbon sources, preferably from 2 to 10 carbon sources, more preferentially 2 to 5 carbon sources and even more preferentially 2 carbon sources.
According to one embodiment, the copolymer according to the invention is derived from several carbon sources and at least one is saturated aliphatic. According to a particular embodiment of the invention, the PHA copolymer(s) consist of two different repeating polymer units chosen from the units (A) and (B) as defined previously.
According to a particular embodiment of the invention, the PHA copolymer(s) contain or preferably consist of two different repeating polymer units chosen from the units (A) as defined previously, the units (B) such that R2 represents a cyclic or non-cyclic, linear or branched, saturated or unsaturated hydrocarbon-based group comprising from 3 to 30 carbon atoms; in particular chosen from linear or branched (C3-C23)alkyl and linear or branched (C3-C23)alkenyl, in particular a linear hydrocarbon-based group, more particularly (C4-C20)alkyl or (C4-C20)alkenyl; preferably, the hydrocarbon-based group has a carbon number corresponding to the number of carbon atoms of the radical R1 from which at least one carbon atom is subtracted, preferably corresponding to the number of carbon atoms of the radical R1 from which two carbon atoms are subtracted.
More particularly, the PHA copolymer(s) according to the invention comprise the repeating unit of formula (I), and also the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof such as hydrates:
According to a particular embodiment, the PHA copolymer(s) of composition a) contain three different repeating polymer units (A), (B) and (C), and preferably consist of three different polymer units (A), (B) and (C) below, and also the optical or geometrical isomers thereof and the solvates thereof such as hydrates:
-[—O—CH(R1)—CH2—C(O)—]- unit (A)
-[—O—CH(R2)—CH2—C(O)—]- unit (B)
-[—O—CH(R3)—CH2—C(O)—]- unit (C)
According to a particular embodiment of the invention, the PHA copolymer(s) comprise the repeating unit of formula (II), and also the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof such as hydrates:
According to a particular embodiment, the PHA copolymer(s) of composition a) contain four different repeating polymer units (A), (B), (C) and (D), and preferably consist of four different polymer units (A), (B), (C) and (D), below, and also the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof such as hydrates:
-[—O—CH(R1)—CH2—C(O)—]- unit (A)
-[—O—CH(R2)—CH2—C(O)—]- unit (B)
-[—O—CH(R3)—CH2—C(O)—]- unit (C)
-[—O—CH(R4)—CH2—C(O)—]- unit (D)
According to a particular embodiment of the invention, the PHA copolymer(s) comprise the repeating unit of formula (III), and also the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof such as hydrates:
in which formula (III):
According to one embodiment, the PHA copolymer(s) of composition a) more particularly contain five different repeating polymer units (A), (B), (C), (D) and (E), and preferably consist of five different polymer units (A), (B), (C), (D) and (E), below, and also the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and also the solvates thereof such as hydrates:
-[—O—CH(R1)—CH2—C(O)—]- unit (A)
-[—O—CH(R2)—CH2—C(O)—]- unit (B)
-[—O—CH(R3)—CH2—C(O)—]- unit (C)
-[—O—CH(R4)—CH2—C(O)—]- unit (D)
-[—O—CH(R5)—CH2—C(O)—]- unit (E)
in which polymer units (A), (B), (C), (D) and (E):
According to a particular embodiment of the invention, the PHA copolymer(s) comprise the repeating unit of formula (IV), and also the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof such as hydrates:
Preferably, R1 represents a linear or branched, preferably linear, (C5-C2)alkyl hydrocarbon-based chain. According to one embodiment of composition C1 according to the invention, the PHA copolymer(s) are such that the radical R1 is an alkyl group comprising 5 to 14 and preferably between 6 and 12 carbon atoms, more preferentially between 7 and 10 carbon atoms such as n-pentyl, n-hexyl, n-octyl or n-nonyl.
According to a particular embodiment of the invention, the hydrocarbon-based chain R1 is unsubstituted. According to a particular embodiment of the invention, the hydrocarbon-based chain R1 is uninterrupted.
According to another embodiment, the hydrocarbon-based chain of the radical R1 of the invention is 1) either substituted, 2) or interrupted, 3) or substituted and interrupted.
According to a particular embodiment of the invention, the PHA copolymer(s) are such that R1 represents a hydrocarbon-based chain, notably an alkyl group as defined previously, which is interrupted with one or more (preferably one) atoms or groups chosen from O, S, N(Ra) and carbonyl, or combinations thereof such as ester, amide or urea, with Ra being as defined previously, preferably Ra represents a hydrogen atom; preferably, R1 represents an alkyl group which is interrupted with one or more atoms chosen from O and S, more preferentially with an O or S, notably S, atom. In particular, when it represents an interrupted hydrocarbon-based chain, notably alkyl, R1 is C7-C20, more particularly C8-C18 and even more particularly C9-C16. Preferably, said interrupted hydrocarbon-based chain, notably alkyl, is linear.
According to another embodiment of the invention, the PHA copolymer(s) are such that R1 represents a hydrocarbon-based chain, notably an alkyl group as defined previously, substituted with one or more (preferably one) atoms or groups chosen from: a) to k) as defined previously. Preferably, said hydrocarbon-based chain is substituted with only one atom or group chosen from: a) to k) as defined previously.
According to a particular embodiment of the invention, the PHA copolymer(s) are such that R1 represents a hydrocarbon-based chain, notably an alkyl group as defined previously, which is substituted with one or more (preferably one) groups chosen from a) halogen such as chlorine or bromine, b) hydroxyl, c) thiol, d) (di)(C1-C4)(alkyl)amino and preferably amino, e) carboxyl, i) (hetero)cycloalkyl such as anhydride, dithiolane or epoxide, j) a cosmetic active agent chosen from coloured or uncoloured, fluorescent or non-fluorescent chromophores such as optical brighteners, UV-screening agents, h) (hetero)aryl such as phenyl or furyl, k) R—X with R representing a group chosen from α) cycloalkyl such as cyclohexyl, β) heterocycloalkyl such as a sugar radical, preferably a monosaccharide such as glucosyl, γ) (hetero)aryl such as phenyl, δ) a cosmetic active agent as defined previously, m) thiosulfate and X representing a′) O, S, N(Ra), b′) carbonyl, c′) or combinations thereof of a′) with b′) such as ester, amide or urea; Ra represents a hydrogen atom or a (C1-C4)alkyl or aryl(C1-C4)alkyl group such as benzyl, preferably Ra represents a hydrogen atom.
Even more preferentially, the PHA copolymer(s) are such that R1 represents a hydrocarbon-based chain, notably an alkyl group as defined previously, which is substituted with one or more (preferably one) groups chosen from a) halogen such as chlorine or bromine, b) hydroxyl, d) (di)(C1-C4)(alkyl)amino, preferably amino, e) carboxyl, i) (hetero)cycloalkyl such as epoxide, h) (hetero)aryl such as phenyl or furyl, k) R—X with R representing a group chosen from α) cycloalkyl such as cyclohexyl, β) heterocycloalkyl such as a sugar radical, preferably a monosaccharide such as glucosyl, γ) (hetero)aryl such as phenyl, and X representing a′) O, S or N(Ra), preferably S; Ra representing a hydrogen atom or a (C1-C4)alkyl group, preferably Ra represents a hydrogen atom.
According to one embodiment, said substituted hydrocarbon-based chain, notably alkyl, is linear.
According to another embodiment, said substituted hydrocarbon-based chain, notably alkyl, is branched.
According to another particular embodiment of the invention, the hydrocarbon-based chain of the radical R1 of the invention is substituted and interrupted.
According to a particular embodiment of the invention, the hydrocarbon-based chain (notably an alkyl group as defined previously) of the radical R1 of the invention is:
According to a preferred embodiment of the invention, the hydrocarbon-based chain (notably an alkyl group as defined previously) of the radical R1 of the invention is:
Preferably, said substituted and interrupted hydrocarbon-based chain is notably alkyl, and is preferably linear.
More preferentially, when said hydrocarbon-based chain R1 is substituted, it is substituted at the end of the chain on the opposite side from the carbon atom which bears said radical R1.
According to one embodiment of the invention, said hydrocarbon-based chain R1 has the following formula —(CH2)rX-(ALK)u-G with X being as defined previously, in particular representing O, S or N(Ra), preferably S, ALK represents a linear or branched, preferably linear, (C1-C10)alkylene and more particularly (C1-C8)alkylene chain, r represents an integer inclusively between 6 and 11, preferably between 7 and 10 such as 8; u is equal to 0 or 1; and G represents a hydrogen atom or a group chosen from hydroxyl, carboxyl, (di)(C1-C4)(alkyl)amino, (hetero)aryl in particular aryl such as phenyl, cycloalkyl such as cyclohexyl, or a sugar, in particular a monosaccharide optionally protected with one or more groups such as acyl, preferably Sug.
According to another particular embodiment of the invention, the PHA copolymer(s) are such that R1 represents (C5-C23)alkyl substituted with one or more halogen atoms such as fluorine, chlorine or bromine, more particularly linear (C5-C20)alkyl, even more particularly (C5-C13)alkyl, substituted with a halogen atom such as bromine. Preferably, the halogen atom is substituted at the end of said alkyl group. More preferentially, R1 represents 1-halo-5-yl such as 1-bromo-5-yl.
According to another particular embodiment of the invention, the PHA copolymer(s) are such that R1 represents a (C5-C23)alkyl group substituted with one or more groups chosen from g) cyano, and more particularly represents a (C3-C13)alkyl group, which is preferably linear, substituted with a cyano group g), such as 1-cyano-3-propyl.
According to another particular embodiment of the invention, the PHA copolymer(s) are such that R1 represents vii) a (hetero)aryl(C1-C2)alkyl and more particularly aryl(C1-C2)alkyl group, preferably phenylethyl.
According to another particular embodiment of the invention, the PHA copolymer(s) are such that R1 represents a (C5-C23)alkyl group substituted with one or more groups chosen from c) (hetero)cycloalkyl. More particularly, R1 represents a (C5-C13)alkyl group, which is preferably linear, substituted with a heterocycloalkyl group such as epoxide or dithiolane, preferably epoxide.
In particular, the PHA copolymer(s) are such that R2 is chosen from linear or branched (C5-C23)alkyl, and linear or branched (C5-C23)alkenyl, in particular a linear hydrocarbon-based group, more particularly (C5-C20)alkyl or (C5-C20)alkenyl, preferably linear or branched, and more particularly linear, (C5-C20)alkyl.
In particular, the PHA copolymer(s) are such that R2 is chosen from linear or branched (C1-C28)alkyl, and linear or branched (C2-C28)alkenyl, in particular a linear hydrocarbon-based group, more particularly (C3-C20)alkyl or (C3-C20)alkenyl; preferably, the hydrocarbon-based group has a carbon number corresponding to the number of carbon atoms of the radical R1 from which at least one carbon atom is subtracted, preferably corresponding to the number of carbon atoms of the radical R1 from which two carbon atoms are subtracted.
According to one embodiment of the invention, the PHA copolymer(s) are such that the radical R2 is a linear or branched, preferably linear, (C3-C8)alkyl, in particular (C3-C6)alkyl, preferably (C4-C6)alkyl group such as n-pentyl or n-hexyl.
According to another embodiment of composition C1 according to the invention, the PHA copolymer(s) comprise a branched (C3-C8)alkyl, particularly (C4-C6)alkyl radical R2 preferably a branched (C4-C5)alkyl radical such as isobutyl.
According to another embodiment of composition C1 according to the invention, the PHA copolymer(s) of the invention comprise the units (A) bearing an alkyl radical R1 as defined previously, the units (B) as defined previously and the units (C) bearing a linear or branched (C6-C20)alkenyl, particularly (C7-C14)alkenyl and more particularly (C8-C10)alkenyl radical, which is preferably linear and comprising only one unsaturation at the chain end, in particular, —[CR4(R5)]q—C(R6)═C(R7)—R8 with R4, R5, R6, R7 and R8, which may be identical or different, representing a hydrogen atom or a (C1-C4)alkyl group such as methyl, preferably a hydrogen atom, and q represents an integer inclusively between 2 and 20, preferably between 3 and 10, more preferentially between 4 and 8 such as 6, such as —[CH2]q—CH═CH2 and q represents an integer inclusively between 3 and 8, preferably between 4 and 6, such as 5.
According to one embodiment of composition C1 according to the invention, the PHA copolymer(s) comprise units (A) bearing an alkyl radical R1 comprising between 8 and 16 carbon atoms substituted with one or more (preferably one) groups chosen from hydroxyl, (di)(C1-C4)(alkyl)amino, carboxyl, and R—X— as defined previously, preferably R—S— with R representing a cycloalkyl group such as cyclohexyl, heterocycloalkyl such as a sugar, more preferentially a monosaccharide such as glucose, optionally substituted aryl(C1-C4)alkyl such as (C1-C4)(alkyl)benzyl or phenylethyl, or heteroaryl(C1-C4)alkyl such as furylmethyl.
According to one embodiment of composition C1′ according to the invention, the copolymer(s) comprise units (B) bearing a linear or branched, preferably linear, (C1-C8)alkyl, particularly (C2-C6)alkyl, preferably (C4-C5)alkyl radical R2 such as pentyl.
According to another embodiment of composition C1 according to the invention, the PHA copolymer(s) comprise units (A) containing an alkyl radical R1 as defined previously, units (B) as defined previously and units (C) containing a linear or branched (C6-C20)alkenyl, particularly (C7-C14)alkenyl radical and more particularly (C8-C10)alkenyl radical, which is preferably linear, and comprising only one unsaturation at the chain end such as —[CH2]q—CH═CH2 and p represents an integer inclusively between 3 and 8, preferably between 4 and 6, such as 5.
According to a particular embodiment of the invention, in the PHA copolymer(s), the unit (A) comprises a hydrocarbon-based chain as defined previously, in particular ii), said unit (A) preferably being present in a molar percentage ranging from 0.1% to 99%, more preferentially a molar percentage ranging from 0.5% to 50%, even more preferentially a molar percentage ranging from 1% to 40%, better still a molar percentage ranging from 2% to 30%.
According to a particular embodiment of the invention, in the PHA copolymer(s), the unit (A) is preferably present in a molar percentage ranging from 0.5% to 99%.
According to one embodiment, when R1 represents a (C5-C28)alkyl group, the unit (A) is preferably present in a molar percentage ranging from 0.5% to 99%, more preferentially from 50% to 99%, more particularly from 60% to 99% and even more preferentially from 70% to 99%. According to this embodiment, the unit (B) is preferably present in a molar percentage ranging from 0.5% to 40%, more preferably from 2% to 40%; and the unit (C) is preferably present in a molar percentage ranging from 0.5% to 20% relative to all the units (A), (B) and (C).
According to another embodiment, when R1 represents a hydrocarbon-based chain chosen from i) linear or branched (C5-C23)alkyl, ii) linear or branched (C5-C23)alkenyl, iii) linear or branched (C5-C23)alkynyl, preferably the hydrocarbon-based group is linear, said hydrocarbon-based chain being substituted with one or more atoms or groups a) to m) and/or interrupted with one or more heteroatoms or groups a′) to c′) as defined for R1; it in particular represents a hydrocarbon-based group chosen from linear or branched (C4-C23)alkyl, optionally substituted with one or more atoms or groups a) to m) and/or interrupted with one or more heteroatoms or groups a′) to c′) as defined previously, the unit (A) is preferably present in a molar percentage ranging from 0.5% to 99%, more preferentially a molar percentage ranging from 1% to 50%, even more preferentially a molar percentage ranging from 2% to 40%, better still a molar percentage ranging from 5% to 30%; the unit (B) is present in a molar percentage ranging from 0.5% to 99.5%, more preferably from 0.5% to 99%, preferentially from 1% to 99%, more preferentially from 1% to 90%, even more preferentially from 2% to 70%; and the unit (C) is present in a molar percentage ranging from 0.5% to 20% relative to all the units (A), (B) and (C). Advantageously, the PHA copolymer(s) of the invention comprise from 2 mol % to 70 mol % of units (B); and from 0.5 mol % to 10 mol % of units (C); more advantageously, the copolymer comprises from 50 mol % to 95 mol % of units (B), and from 0.5 mol % to 7 mol % of units (C).
According to a more particular embodiment of the invention, the PHA copolymer(s) are such that, in the PHA copolymer(s) a):
Preferably, when R1 of the unit (A) comprises a saturated unsubstituted and uninterrupted hydrocarbon-based chain, said unit (A) is present in a molar percentage of greater than 30%, more particularly greater than 50%, more preferentially greater than 60%, preferably between 60% and 90%.
The values of the molar percentages of the units (A), (B) and (C) of the PHA copolymer(s) are calculated relative to the total number of moles of (A)+(B) if the copolymer(s) do not comprise any additional units (C); otherwise, if the copolymer(s) of the invention contain three different units (A), (B) and (C), then the molar percentage is calculated relative to the total number of moles (A)+(B)+(C); otherwise, if the copolymer(s) of the invention contain four different units (A), (B), (C) and (D), then the molar percentage is calculated relative to the total number of moles (A)+(B)+(C)+(D); otherwise, if the copolymer(s) of the invention contain five different units (A), (B), (C), (D) and (E), then the molar percentage is calculated relative to the total number of moles (A)+(B)+(C)+(D)+(E).
According to a particular form of the invention, the unit(s) (A) of the PHA copolymer(s) are chosen from the following repeating units (A), and also the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof such as hydrates:
in which repeating units A1 to A12:
In particular, the stereochemistry of the carbon atoms bearing the radicals R1 is of (R) configuration.
According to one form of the invention, the PHA copolymer(s) of the invention comprise the repeating units (B) of formula (A12), and also the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof such as hydrates, it being understood that (B) is different from (A).
Preferentially, the PHA copolymer(s) of the invention comprise the following repeating units, and also the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof such as hydrates:
with Re representing a group Rf—C(O)—, with Rf representing a (C1-C4) alkyl group such as methyl.
In particular, the stereochemistry of the carbon atoms bearing the radicals R1 and R2 is of the same (R) or (S) configuration, preferably of (R) configuration.
More particularly, the stereochemistry of the carbon atoms bearing the radicals R1, R2 and R3 is of the same (R) or (S) configuration, preferably of (R) configuration. More particularly, the stereochemistry of the carbon atoms bearing the radicals R1, R2, R3 and R4 is of the same (R) or (S) configuration, preferably of (R) configuration.
More particularly, the stereochemistry of the carbon atoms bearing the radicals R1, R2, R3, R4 and R5 is of the same (R) or (S) configuration, preferably of (R) configuration.
More preferentially, the PHA copolymer(s) have the following formula, and also the optical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof such as hydrates:
m, n, Hal, t, Ar, Ar′, Cycl, Fur and Sug are as defined previously for compounds (1) to (14).
)
—CH═CH2
—CH═CH2
)
—CH═CH2
—CH3
)2—CH3
—CH3
—CH3
—CH3
)4—CH3
—S—
—S—(CH2)7—CH3
)—C(O)—OH
—S—(CH2)7—CH3
)8—S—
—S—(CH3)
—OH
)4—S—(CH2)8—OH
—S—
—S—(CH
)
—NH2
—NH
)2—NH2
—S—Cyol
—CH3
indicates data missing or illegible when filed
Preferably, the PHA(s) of the invention are chosen from compounds (15), (16) and (17), in particular (16).
More particularly, the PHA(s) of the invention are chosen from compounds (15′), (16′) and (17′), in particular (16′).
More particularly, the PHA a) of the invention is compound (23′).
Preferably, the PHA(s) a) of the invention are chosen from compounds (25), (26), (31) and (32), in particular (26).
According to a particularly preferred embodiment of the invention, the PHA polymer(s) a) are chosen from the polyhydroxyalkanoate (PHA) copolymers of examples 1, 11, 12, 21 and 25 as described thereafter; more preferentially from examples 1d, 11′, 12, 21 and 25 as described thereafter.
The PHA copolymer(s) of the invention preferably have a number-average molecular weight ranging from 50 000 to 150 000.
The molecular weight may notably be measured by size exclusion chromatography. A method is described below in the examples.
Preferably, the PHA copolymer(s) are present in composition C1 according to the invention in a content ranging from 0.1% to 65% by weight; more preferably from 0.1% to 60% by weight, preferentially from 1% to 50% by weight; more preferentially from 3% to 40% by weight, even more preferentially from 5% to 35% by weight, better from 10% to 30%, and even better from 15% to 20% by weight, relative to the total weight of the composition.
According to a variant of the present invention, the composition, preferably cosmetic, comprises:
-[—O—CH(R1)—CH2—C(O)—]- unit (A)
The methods for preparing the PHA copolymer(s) of the invention are known to those skilled in the art. Mention may notably be made of the use of “functionalizable” PHA-producing microbial strains.
The term “functionalizable” means that the PHA copolymer(s) comprise a hydrocarbon-based chain comprising one or more atoms or groups that are capable of reacting chemically with another reagent—also referred to as “reactive atoms or reactive groups”—to give a Σ covalent bond with said reagent. The reagent is, for example, a compound comprising at least one nucleophilic group and said functionalized hydrocarbon-based chain comprises at least one electrophilic or nucleofugal atom or group, the nucleophilic group(s) reacting with the electrophilic group(s) to covalently graft Σ the reagent. The nucleophilic reagent may also react with one or more unsaturations of the alkenyl group(s) to also lead to grafting by covalent bonding of the functionalized hydrocarbon-based chain with said reagent. The addition reaction may also be radical-based, an addition of Markovnikov or anti-Markovnikov type, or nucleophilic or electrophilic substitution. The addition or condensation reactions may or may not take place via a radical route, with or without the use of catalysts or of enzymes, with heating preferably to a temperature less than or equal to 100° C. or without supplying heat, under a pressure of greater than 1 atm or otherwise, under an inert atmosphere or otherwise, or under oxygen or otherwise.
The term “nucleophilic” refers to any atom or group which is electron-donating by an inductive effect +I and/or a mesomeric effect +M. Electron-donating groups that may be mentioned include hydroxyl, thiol and amino groups.
The term “electrophilic” refers to any atom or group which is electron-withdrawing by an inductive effect −I and/or a mesomeric effect −M.
The microorganisms producing PHAs of the invention notably bearing a hydrocarbon-based chain may be naturally produced by the bacterial kingdom, such as Cyanobacteria of the order of Nostocales (e.g.: Nostoc muscorum, Synechocystis and Synechococcus) but mainly by the Proteobacteria, for example in the class of:
Among the microorganisms of the bacterial kingdom, the genera Azotobacter, Hydrogenomomas or Chromatium are the most representative of the PHA-producing organisms.
The organisms which naturally produce PHAs bearing notably a C3-C5 hydrocarbon-based chain are notably Proteobacteria, such as gamma-Proteobacteria, and more particularly of the order Pseudomonales of the family Pseudomonas such as Pseudomonas resinovorans, Pseudomonas putida, Pseudomonas fluorescens, Pseudomonas aeruginosa, Pseudomonas citronellolis, Pseudomonas mendocina, Pseudomonas chlororaphis; preferably Pseudomonas putida and more preferentially Pseudomonas putida GPo1 and Pseudomonas putida KT2440.
Certain organisms may also naturally produce PHAs without belonging to the order of Pseudomonales, such as Commamonas testosteroni which belongs to the class of beta-Proteobacteria of the order Burkholderiales of the family of Comamonadaceae.
The microorganism producing PHAs according to the invention may also be a recombinant strain if a 3-oxidation PHA synthase metabolic pathway is present. The 3-oxidation PHA synthase metabolic pathway is mainly represented by four classes of enzymes, EC: 2.3.1 B2, EC: 2.3.1 B3, EC: 2.3.1 B4 and EC: 2.3.1 B5.
The recombinant strain may be from the Bacteria kingdom, for instance Escherichia coli, or from the Plantae kingdom, for instance Chlorella pyrenoidosa (International Journal of Biological Macromolecules, 116, 552-562 “Influence of nitrogen on growth, biomass composition, production, and properties of polyhydroxyalkanoates (PHAs) by microalgae”) or from the Fungi kingdom, for instance Saccharomyces cerevisiae or Yarrowia lipolytica: Applied Microbiology and Biotechnology 91, 1327-1340 (2011) “Engineering polyhydroxyalkanoate content and monomer composition in the oleaginous yeast Yarrowia lipolytica by modifying the β-oxidation multifunctional protein”).
Use may also be made of genetically modified microorganisms, which may make it possible, for example, to increase the production of PHA, and/or to increase the oxygen consumption capacity, and/or to reduce the autolysis and/or to modify the monomer ratio.
It is known that, for PHAs, a large portion of the total production cost is devoted to the culture medium and mainly to the substrate/carbon source. Use may thus be made of genetically modified microorganisms using a smaller amount of nutrient (carbon source) for their growth, for example microorganisms that are photo-autotrophic by nature, i.e. using light and CO2 as main energy source.
The copolymer may be obtained in a known manner by biosynthesis, for example with the microorganisms belonging to the genus Pseudomonas, such as Pseudomonas resinovorans, Pseudomomonas putida, Pseudomonas fluorescens, Pseudomonas aeruginosa, Pseudomonas citronellolis, Pseudomonas mendocina, Pseudomonas chlororaphis and preferably Pseudomonas putida; and with a carbon source which may be a C2-C20, preferably C6-C18, carboxylic acid, such as acetic acid, propionic acid, butyric acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, dodecanoic acid, or an alkenoic acid such as undecylenic acid; a saccharide, such as fructose, maltose, lactose, xylose, arabinose, etc.); an n-alkane, such as hexane, octane or dodecane; an n-alcohol, such as methanol, ethanol, octanol or glycerol; methane or carbon dioxide.
The biosynthesis may optionally be performed in the presence of an inhibitor of the β-oxidation pathway, such as acrylic acid, methacrylic acid, propionic acid, cinnamic acid, salicylic acid, pentenoic acid, 2-butynoic acid, 2-octynoic acid or phenylpropionic acid, and preferably acrylic acid.
According to one embodiment, the process for preparing the PHAs of the invention uses microbial cells which produce PHAs via genetically modified microorganisms (GMOs). The genetic modification may increase the production of PHA, increase the oxygen absorption capacity, increase the resistance to the toxicity of solvents, reduce the autolysis, modify the ratio of the PHA comonomers, and/or any combination thereof. In some of these embodiments, the modification of the comonomer ratio of the unit (A) increases the amount of predominant monomer versus (B) of the PHA of the invention which is obtained. In another embodiment, the PHA-producing microbial cells reproduce naturally.
By way of example, a genetically modified microbial strain producing PHA that is functionalizable or comprising a reactive group that may be mentioned is Pseudomonas entomophila LAC23 (Biomacromolecules. 2014 Jun. 9; 15(6):2310-9. doi: 10.1021/bm500669s).
It is also possible to use genetically modified microorganisms which produce phenylvaleric-co-3-hydroxydodecanoic copolymers (Sci. China Life Sci., Shen R., et al., 57, No. 1, (2014) with a strain such as Pseudomonas entomophila LAC23.
Nutrients, such as water-soluble salts based on nitrogen, phosphorus, sulfur, magnesium, sodium, potassium and iron, may also be used for the biosynthesis.
The appropriate known conditions of temperature, pH and dissolved oxygen (OD) can be used for the culturing of the microorganisms.
The microorganisms may be cultured according to any known method of culturing, such as in a bioreactor in continuous or batch mode, in fed or unfed mode.
The biosynthesis of the polymers used according to the invention is notably described in the article “Biosynthesis and Properties of Medium-Chain-Length Polyhydroxyalkanoates with Enriched Content of the Dominant Monomer”, Xun Juan et al., Biomacromolecules 2012, 13, 2926-2932, and in patent application WO 2011/069244.
The microbial strains producing PHA which is functionalizable or comprising a reactive group, as defined previously, are, for example, of the genus Pseudomonas such as P. cichorii YN2, P. citronellolis, P. jessenii, and more generally with species of Pseudomonas putida such as Pseudomonas putida GPo1 (synonym of Pseudomonas oleovorans), P. putida KT2442, P. putida KT2440, P. putida KCTC 2407 and P. putida BM01, and in particular P. putida KT2440.
One means for gaining access to the PHAs of the invention is to introduce one or more organic compounds into the culture medium, this or these organic compounds representing one or more carbon sources preferably chosen from alkanes, alkenes, alcohols, carboxylic acids and a mixture thereof.
In one embodiment, the organic compound(s) will preferably be chosen from alcohols, carboxylic acids and a mixture thereof.
The carbon source(s) may be classified in two categories:
1) Carbon Source Via One or More Organic Compounds Introduced into the Medium:
According to a particular embodiment of the invention, the organic compound(s) are chosen from alcohols, in particular (C5-C20)alkanols, and/or carboxylic acids, in particular optionally substituted and/or interrupted (C5-C20)alkanoic acids, notably (C5-C20)alkanoic acids such as (C7-C11)alkanoic acids, for instance nonanoic acid or pelargonic acid and/or (C5-C20)alkenoic acids, notably (C5-C20)alkenoic acids such as (C7-C11)alkenoic acids, for instance undecylenic acid, and mixtures thereof.
The carbon source(s) may be classified into three groups according to their intended use:
Such microbiological processes are known to those skilled in the art, notably in the scientific literature. Mention may be made of: International Journal of Biological Macromolecules 28, 23-29 (2000); The Journal of Microbiology, 45, No. 2, 87-97, (2007).
According to one variant, the integration of the substrate that is structurally linked to the reactive atom(s) or to the reactive group(s) of the PHA(s) of the invention is introduced directly into the medium as sole carbon source in a medium suitable for microbial growth. (Example: group A for P. putida GPo1: alkenoic acid, notably terminal).
According to another variant, the integration of the substrate that is structurally linked to the reactive atom(s), notably halogen, or to the reactive group(s) of the PHA(s) of the invention is introduced into the medium as carbon source with a second carbon source as co-substrate which is also structurally linked to the PHA, in a medium suitable for microbial growth. (Example: group B for P. putida GPo1: haloalkanoic acids which are preferably terminal, such as terminal bromoalkanoic acids).
According to yet another variant, the integration of the substrate that is structurally linked to the reactive atom(s), notably halogen, or to the reactive group(s) of the PHA(s) of the invention may be introduced directly into the medium as carbon source with a second carbon source as co-substrate which is also structurally linked to the PHA(s) and a third carbon source as co-substrate which is not structurally linked to the PHA(s), in a medium suitable for microbial growth. (Example: group C glucose or sucrose).
In one embodiment, the β-oxidation pathway inhibitor is acrylic acid, 2-butynoic acid, 2-octynoic acid, phenylpropionic acid, propionic acid, trans-cinnamic acid, salicylic acid, methacrylic acid, 4-pentenoic acid or 3-mercaptopropionic acid, preferably acrylic acid.
In one embodiment of the first aspect, the functionalized fatty acid is a functionalized hexanoic acid, functionalized heptanoic acid, functionalized octanoic acid, functionalized nonanoic acid, functionalized decanoic acid, functionalized undecanoic acid, functionalized dodecanoic acid or functionalized tetradecanoic acid.
The functionalization may be introduced by means of an organic compound chosen from precursors of the alcohol and/or carboxylic acid category, notably:
The review International Microbiology 16:1-15 (2013) doi:10.2436/20.1501.01.175 also mentions the majority of the functionalized native PHAs.
In a particular embodiment of the invention, the fatty acid from group A is chosen from 11-undecenoic acid, 10-epoxyundecanoic acid, 5-phenylvaleric acid, citronellol and 5-cyanopentanoic acid.
In a particular embodiment of the invention, the fatty acid from group A is chosen from halooctanoic acids such as 8-bromooctanoic acid.
In a particular embodiment of the invention, the carbon source from group C is a monosaccharide, preferably glucose.
2) Carbon Source in the Presence of Oxidation Inhibitor Introduced into the Medium:
Another aspect of the invention is the use of the PHA-producing microbial strains in a medium that is suitable for microbial growth, said medium comprising: a substrate which is structurally linked to the PHA(s); at least one carbon source which is not structurally linked to the PHA(s); and at least one oxidation and notably β-oxidation pathway inhibitor. This allows the growth of the microbial cells to take place in said medium, the microbial cells synthesizing the PHA polymer(s) of the invention; preferably copolymer particularly containing more than 95% of identical units, which has a comonomer ratio of unit (A) and of unit (B) which differs from that obtained in the absence of the β-oxidation pathway inhibitor.
The scheme below illustrates, by way of example, the functionalization of PHA copolymers according to the invention starting from a PHA copolymer bearing an unsaturated hydrocarbon-based chain, according to Scheme 1 below:
in which Scheme 1:
Other reactions may be performed using double or triple unsaturations such as Michael or Diels-Alder additions, radical reactions, catalytic (notably with Pd or Ni) or non-catalytic hydrogenation reactions, halogenation reactions, notably with bromine, hydration reactions or oxidation reactions, which may or may not be controlled, and reactions on electrophiles as represented schematically below.
According to a particular embodiment of the invention, the PHA copolymers comprise
In particular, the hydrothiolation reactions may be performed in the presence of a thermal initiator, a redox initiator or a photochemical initiator and of an organic compound bearing a sulfhydryl group, notably chosen from:
Examples of initiators that may be mentioned include: tert-butyl peroxy-2-ethylhexanoate, cumene perpivalate, tert-butyl peroxylaurate, benzoyl peroxide, lauroyl peroxide, octanoyl peroxide, di-tert-butyl peroxide, tert-butylcumyl peroxide, dicumyl peroxide, 2,2′-azobisisobutyronitrile, 2,2′-azobis(2-methylbutyronitrile), 2,2′-azobis(2,4-dimethylvaleronitrile), 2,2′-azobis(4-methoxy-2,4-dimethylvaleronitrile), 1,1-bis(tert-butylperoxy)-3,3,5-trimethylcyclohexane, 1,1-bis(tert-butylperoxy)cyclohexane, 1,4-bis(tert-butylperoxycarbonyl)cyclohexane, 2,2-bis(tert-butylperoxy)octane, n-butyl 4,4-bis(tert-butylperoxy)valerate, 2,2-bis(tert-butylperoxy)butane, 1,3-bis(tert-butylperoxyisopropyl)benzene, 2,5-dimethyl-2,5-bis(tert-butylperoxy)hexane, 2,5-dimethyl-2,5-bis(benzoylperoxy)hexane, di-tert-butyl diperoxyisophthalate, 2,2-bis(4,4-di-tert-butylperoxycyclohexyl)propane, di-tert-butyl peroxy-α-methylsuccinate, di-tert-butyl peroxydimethylglutarate, di-tert-butyl peroxyhexahydroterephthalate, di-tert-butyl peroxyazelate, 2,5-dimethyl-2,5-bis(tert-butylperoxy)hexane, diethylene glycol bis(tert-butylperoxycarbonate), di-tert-butyl peroxytrimethyladipate, tris(tert-butylperoxy)triazine, vinyltris(tert-butylperoxy)silane phenothiazine, tetracene, perylene, anthracene, 9,10-diphenylanthracene, thioxanthone, benzophenone, acetophenone, xanthone, fluorenone, anthraquinone, 9,10-dimethylanthracene, 2-ethyl-9,10-dimethyloxyanthracene, 2,6-dimethylnaphthalene, 2,5-diphenyl-1,3,4-oxadiazole, xanthopinacol, 1,2-benzanthracene, 9-nitroanthracene. Each of these initiators may be used alone or in combination with others.
The chemical reactions mentioned previously are known to those skilled in the art. Mention may notably be made of the following documents: Synthesis and preparation of PHAs modified with polyethylene glycol dithiol: 10.1021/acs.biomac.9b00479; Biomacromolecules, 19, 3536-3548 (2018); Synthesis and preparation of PHAs modified with mercaptohexanol: 10.1021/acs.biomac.8b01257; Biomacromolecules, 20, 2, 645-652 (2019); Synthesis and preparation of PHAs modified with hydroxycinnamic acid sulfate, and zosteric acid: 10.1021/bm049962e; Biomacromolecules, 5, 4, 1452-1456 (2004); Radical addition of methyl methacrylate to a PHOUn: 10.1002/1521-3935(20010701)202:11<2281::AID-MACP2281>3.0.CO;2-9; Macromolecular Chemistry and Physics, vol. 202, 11, 2281-2286 (2001); Synthesis and preparation of PHAs modified with a polysilsesquioxane (POSS): 10.1016/j.polymer.2005.04.020; Polymer Vol. 46, 14, 5025-5031 (2005); Grafting of thio-beta-glucose onto unsaturated side chains: 1022-1336/99/0202-0091$17.50+0.50/0; Macromol. Rapid Commun., 20, 91-94 (1999); and/or
The chemical reactions mentioned previously are known to those skilled in the art. Mention may notably be made of the following documents: 10.1021/bm049337; Biomacromolecules, vol. 6, 2, 891-896 (2005); 10.1016/S0032-3861(99)00347-X; Polymer, vol. 41, 5, 1703-1709 (2000); 10.1021/ma9714528 and 10.1016/S1381-5148(97)00024-2; Macromolecules, 23, 15, 3705-3707 (1990); 10.1016/S0032-3861(01)00692-9; Polymer, vol. 43, 4, 1095-1101 (2002); 10.1016/S0032-3861(99)00347-X; Polymer, vol. 41, 5, 1703-1709 (2000); and 10.1021/bm025728h; Biomacromolecules, vol. 4, 2, 193-195 (2003).
Example of functionalization of PHA copolymers according to the invention starting from a PHA copolymer bearing a hydrocarbon-based chain containing an epoxide group, according to Scheme 2 below:
in which Scheme 2 Y, m, n, q′ and R2 are as defined in Scheme 1.
The epoxide structure may be obtained via a conventional method known to those skilled in the art, whether via biotechnological processes or via chemical processes such as oxidation of unsaturation as mentioned previously. The peroxide group(s) may react with carboxylic acids, maleic anhydrides, amines, alcohols, thiols or isocyanates, all these reagents including at least one linear or branched, cyclic or acyclic, saturated or unsaturated C1-C20 hydrocarbon-based chain, or borne by an oligomer or polymer, in particular amino (poly)saccharides such as compounds derived from chitosan and (poly)sil(ox)anes; 3-glycidyloxypropyltrimethoxysilane, 3-aminopropyltriethoxysilane 3-(trimethoxysilyl)propylcarbamic acid, diethanolamine, or 3-mercapto-1-propanesulfonate of alkali metal or alkaline-earth metal salts such as sodium. The epoxide groups may also react with water.
Mention may notably be made of the following documents:
Example of functionalization of PHA copolymers according to the invention starting from a PHA copolymer bearing a hydrocarbon-based chain containing a nucleofugal group, according to Scheme 3 below:
in which Scheme 3 Y, m, n, q′ and R2 are as defined in Scheme 1. M corresponds to an organic or inorganic nucleofugal group, which may be substituted with a nucleophilic group; preferably, said nucleophile is a heteroatom which is electron-donating via the +I and/or +M effect such as 0, S or N. Preferably, the nucleofugal group M is chosen from halogen atoms such as Br, and mesylate, tosylate or triflate groups. This is a reaction known to those skilled in the art. Mention may be made, for example, of the following document: 10.1016/j.ijbiomac.2016.11.118, International Journal of Biological Macromolecules, vol. 95, 796-808 (2017).
Example of functionalization of PHA copolymers according to the invention starting from a PHA copolymer bearing a hydrocarbon-based chain containing a cyano group, according to Scheme 4 below:
in which Scheme 4 Y, m, n, q′ and R2 are ad defined in Scheme 1.
In a first step i), the PHA copolymer bearing a side chain containing a cyano or nitrile group reacts with an organo-alkali metal or organomagnesium compound Y-MgHal, Y—Li or Y—Na, followed by hydrolysis to give the PHA copolymer bearing a side chain containing a group Y grafted with a ketone function. The ketone function may be converted into a thio ketone by thionation, for example with S8 in the presence of amine, or with Lawesson's reagent. Said thio ketone, after total reduction ii) (for example by Clemmensen reduction), leads to the PHA copolymer bearing a side chain containing a group Y grafted with an alkylene group. Alternatively, said thio ketone may undergo a controlled reduction iii) with a conventional reducing agent to give the PHA copolymer bearing a side chain containing a group Y grafted with a hydroxyalkylene group. The cyano group of the starting PHA copolymer can react with water after hydration v) to give the amide derivative, after hydrolysis iv) to the carboxyl derivative. The cyano group of the starting PHA copolymer can also, after reduction vi), give the amine derivative or the ketone derivative. PHA copolymers with a hydrocarbon-based chain bearing a nitrile function are prepared via conventional methods known to those skilled in the art. Mention may be made, for example, of the document: 10.1016/0378-1097(92)90311-B, FEMS Microbiology Letters, vol. 103, 2-4, 207-214 (1992).
Example of functionalization of PHA copolymers according to the invention starting from a PHA copolymer bearing a hydrocarbon-based chain at the chain end, according to Scheme 5 below:
in which Scheme 5 R1, R2, m, n and Y are as defined previously, and R′1 represents a hydrocarbon-based chain chosen from i) linear or branched (C1-C20)alkyl, ii) linear or branched (C2-C20)alkenyl, iii) linear or branched (C2-C20)alkynyl; preferably, the hydrocarbon-based group is linear; said hydrocarbon-based chain being substituted with one or more atoms or groups chosen from: a) halogens such as chlorine or bromine, b) hydroxyl, c) thiol, d) (di)(C1-C4)(alkyl)amino, e) (thio)carboxyl, f) (thio)carboxamide —C(O)—N(Ra)2 or —C(S)—N(Ra)2, f) cyano, g) iso(thio)cyanate, h) (hetero)aryl such as phenyl or furyl, and i) (hetero)cycloalkyl such as anhydride, or epoxide, j) a cosmetic active agent chosen from coloured or uncoloured, fluorescent or non-fluorescent chromophores such as those derived from optical brighteners, or chromophores derived from UVA and/or UVB screening agents, and anti-ageing active agents.
These chain-end grafts onto PHA polymers are known to those skilled in the art. Mention may be made, for example, of the following documents:
Mention may also be made of other methods known to those skilled in the art:
The combination of grafted PHA copolymers of the invention described previously, according to Scheme 6:
in which Scheme 6 R′1, R2, m, n and Y are as defined previously, and X′ represents a reactive atom or group that is capable of reacting with an electrophilic or nucleophilic
atom or group to create a Σ covalent bond; if X′ is an electrophilic or nucleofugal group, then it can react with a reagent R′1-
; if X′ is a nucleophilic group
then it can react with R′1-
to create a Σ covalent bond.
By way of example, the Z covalent bonds or bonding group that may be generated are listed in the table below, from condensation of electrophiles with nucleophiles:
It is also possible, starting with a PHA functionalized on a side chain, to perform chain-end grafting in a second stage as described in Scheme 7. The reverse is also true, in which the chain-end grafting may be performed in a first stage, followed by performing functionalization of a functionalizable side chain in a second stage.
in which Scheme 7 R′1, R2, m, n and Y are as defined previously.
All these chemical reactions are known to those skilled in the art. Mention may be made, for example, of the following documents:
The composition according to the invention comprises one or more oligo/polyester(s) other than the PHA(s) a) as defined previously and the fatty substance(s) c) as defined below.
The term “oligo/polyester(s)” means one or more esters chosen from oligoesters and polyesters preferably of biobased origin.
The term “oligoester” means a hydrocarbon-based chain resulting from the polycondensation of b1) one or more polyol(s) and b2)/b3) with one or more saturated or unsaturated fatty acids comprising one or more carboxyl groups (also called carboxylic acid group —C(O)—OH) and containing between 2 and 10 ester repeating units (ester units also called ester monomers) −[—R—C(O)—O—]n— or R(—[R′C(O)—O]n—R″)m with n and m representing an integer between 2 and 10 with n+m representing an integer between 2 and 10 and R, R′ and R″ representing a linear or branched, cyclic or non-cyclic, saturated or unsaturated, aromatic or non-aromatic hydrocarbon-based chain, which is optionally substituted, notably with one or more hydroxyl or carboxyl groups and/or interrupted with one or more heteroatoms such as oxygen. Oligoesters are short analogues of polyester polymers.
The term “polyester” means a hydrocarbon-based chain, containing more than 11 ester repeating units −[—R—C(O)—O—]n′— or R(—[R′C(O)—O]n—R″)m′ with n‘ and m’ representing an integer greater than or equal to 11 with n′+m′ representing an integer greater than or equal to 11 with R, R′ and R″ as defined previously.
As described previously, the oligo/polyester(s) according to the invention are obtained from at least one polyol component, denoted (b1), preferably comprising from 2 to 6 hydroxyl groups (—OH), in particular three or four hydroxyl groups, preferably 3 OH.
It is understood that the oligo/polyester(s) according to the invention may be prepared from a single polyol or from a mixture of at least two polyols.
In the continuation of the text, unless otherwise specified, the term “polyol” thus means a single polyol compound or a mixture of several polyol compounds, said polyol(s) preferably being organic compounds. Preferably, the polyol component b1) is green, in particular of natural origin, in particular biobased and notably of plant origin.
According to a particular embodiment, the polyol b1) is an organic compound comprising a linear or branched, acyclic or (poly)cyclic, saturated or unsaturated, aromatic or unsaturated hydrocarbon-based chain comprising from 3 to 18 carbon atoms, in particular from 4 to 12 carbon atoms, or even from 5 to 10 carbon atoms; and from 2 to 6 hydroxyl groups, in particular from 3 to 6 hydroxyl groups, said hydrocarbon-based chain being optionally interrupted with one or more heteroatoms, notably of oxygen, in particular possibly bearing ether functions.
As examples of such polyols (al), mention may be made of, without this list being limiting, i) the saccharides as defined previously, notably pentoses such as ribose, arabinose, xylose, lyxose, ribulose, xylulose or hexoses such as allose, altrose, galactose, glucose, idose, mannose, talose, fructose, sorbose or deoxyhexoses such as fucose or rhamnose; ii) triols such as glycerol, iii) tetraols such as pentaerythritol (tetramethylolmethane), erythritol, diglycerol, iv) pentols such as xylitol, v) hexols such as sorbitol and mannitol, or dipentaerythritol or triglycerol, and mixtures thereof.
According to a particular embodiment, the polyol b1) is an oligomer derived from one or more diols of formula (I) HO-ALK-OH, in which formula (I) ALK represents a linear or branched C1-C18, in particular C1-C6, alkylene group, optionally substituted with one or more hydroxyl groups, in particular one hydroxyl group. The polyol b1) is then a poly(C1-C18)alkanediol.
In particular, the polyol may be an oligomer derived from one or more diols of the abovementioned formula (I) with a weight-average molecular mass of between 200 and 4000 g·mol−1, in particular between 300 and 3000 g·mol−1.
By way of example, the compound of formula (I) is such that ALK represents a (C1-C6)alkylene group, the oligomer then being a poly(C1-C6)alkanediol.
Mention may notably be made of polypropanediol (also known as polypropylene glycol) with a weight-average molecular mass of between 300 and 3000 g·mol−1, or polyethylene glycol with a weight-average molecular mass of between 300 and 3000 g·mol−1.
According to another embodiment, the polyol may be chosen from sugars, saccharides and polysaccharides.
According to another preferred embodiment, the oligo/polyester(s) of the invention are derived from condensation with one or more organic polyol(s) b1) chosen from:
According to a particular embodiment, the polyol b1) comprises at least one compound from among glycerol, pentaerythritol and/or sorbitol. In particular, the polyol b1) may be chosen from glycerol, pentaerythritol, sorbitol and mixtures thereof.
According to a particular embodiment, said polyol b1) comprises at least glycerol. In a particular embodiment, the polyol b1) is glycerol.
According to one embodiment, said polyol b1) comprises at least pentaerythritol. In a particular embodiment, the polyol b1) is pentaerythritol.
Said polyol component(s) b1), in particular glycerol and/or pentaerythritol, may represent from 0.1% to 40% by weight, in particular from 1% to 30% by weight, more particularly from 5% to 25% by weight, preferably from 10% to 20% by weight, relative to the total weight of the oligo/polyester(s) according to the invention.
This is the mass proportion of said polyol component(s) b1) relative to the total mass of all the components for the synthesis of oligo/polyester(s).
The content of said polyol(s) b1), in particular glycerol and/or pentaerythritol, in oligo/polyester(s) b) according to the invention may be between 0.1% and 40% by weight, in particular between 1% and 30% by weight, more particularly between 5% and 25% by weight, notably between 5% and 20% by weight, even more particularly between 10% and 15% by weight, relative to the total weight of oligo/polyester(s) according to the invention.
The mass content of said polyol(s) b1), and more generally the mass content of one of the components of the oligo/polyester(s) according to the invention, may be obtained from the amounts of each of the components used in the preparation of said oligo/polyester(s), relative to the mass of oligo/polyester(s).
As described previously, the oligo/polyester(s) of the invention are obtained from at least one polycarboxylic acid, also referred to in the text hereinbelow as “polyacid”, denoted b2).
Said polyacid may be acyclic or (poly)cyclic, saturated or unsaturated, aromatic or non-aromatic, linear or branched, and comprises at least two carboxyl groups —C(O)OH, in particular two to four —C(O)OH groups and more particularly two —C(O)OH groups.
It is understood that oligo/polyester(s) may be prepared from a single polyacid or from a mixture of at least two polyacids. In the continuation of the text, the term “polyacid” or “polycarboxylic acid” means a single polyacid compound or a mixture of several polyacid compounds, unless otherwise specified. Advantageously, said polyacid may be biobased, in particular of plant origin.
Said polycarboxylic acid b2) may be chosen more particularly from linear, branched and/or cyclic, saturated or unsaturated, or even aromatic, polycarboxylic acids comprising from 2 to 54 carbon atoms, notably from 2 to 50 and more particularly from 3 to 40 carbon atoms, such as from 3 to 36 carbon atoms, said acid comprising at least two carboxyl groups —C(O)OH, notably from 2 to 4 carboxyl groups.
In particular, the polycarboxylic acid b2) may comprise two carboxyl groups; it is then referred to as a dicarboxylic acid or diacid.
According to a particular embodiment, said polycarboxylic acid b2) may be chosen from polycarboxylic acids, in particular dicarboxylic acids, having a molecular mass MM of less than or equal to 200 g·mol−1.
In particular, said polycarboxylic acid b2) may be chosen from oxalic acid, succinic acid, adipic acid, and mixtures thereof, and also optical or geometric isomers thereof, organic or mineral base salts thereof, and solvates thereof such as hydrates.
According to another particular embodiment of the invention, the polycarboxylic acid b2) used to form b) according to the invention may be chosen from acids, in particular diacids, with a molecular mass MM of greater than 200 g·mol−1.
In particular, the polycarboxylic acid b2) may be chosen from fatty acids (also called fatty polyacids), in particular dicarboxylic fatty acids (also called fatty diacids), and notably from fatty acid dimers.
The term “fatty acid” means a carboxylic acid comprising at least one linear or branched, preferably linear, saturated or unsaturated aliphatic chain, which may contain at least one carbocycle, in particular one or two carbocycles, notably of the monocyclic or polycyclic cycloalkyl type, for example cyclohexyl or decalinyl (cis or trans decalinyl). The fatty acids are C8 to C38 acids, in particular C8 to C36 acids, notably C10 to C34 acids, it being understood that the number of carbon atoms characterizing a fatty acid takes into account the aliphatic chain of the fatty acid and the carbon atom(s) of said carboxyl group(s). Preferably, the number of carbon atoms of the fatty acid is even.
The dicarboxylic fatty acids may be chosen from the compounds of formula (II) below, and also the salts thereof with organic or mineral bases, and the solvates thereof such as hydrates HO—C(O)-ALK′-C(O)—OH (II) in which ALK′ represents an aliphatic chain as defined previously for the fatty acid, preferably of C16 to C38, notably C32 to C36, in particular C34.
As examples of dicarboxylic fatty acids, mention may be made of sebacic acid and fatty acid dimers.
The term “fatty acid dimer” refers to the product of dimerization of monounsaturated or polyunsaturated fatty acids.
Thus, in a particular embodiment, the polyacid component b2) comprises at least, or even is formed of, a diacid of fatty acid dimer type.
The fatty acids from which fatty acid dimers may be prepared may be chosen more particularly from oleic acid, linoleic acid, palmitoleic acid, linolenic acid, eleostearic acid and mixtures thereof.
The polyacid b2) may be a C18 to C38, in particular C34 to C38 and more particularly C36 fatty acid dimer.
In particular, the fatty acid dimer may be a diacid of formula (II) as defined previously, in which ALK′ represents an aliphatic chain, preferably of C16 to C38, notably C32 to C36, in particular C34, incorporating at least one carbocycle group, preferably one or two carbocycle(s), and more preferentially a central carbocycle group, in particular of the monocyclic or polycyclic cycloalkyl type, in particular cyclohexyl or decalinyl, in particular cyclohexyl.
In a particular embodiment, the polycarboxylic acid b2) may be a diacid of fatty acid dimer type containing a cycloalkyl group, in particular a cyclohexyl group, connected to the —C(O)OH groups via C2 to C10, in particular C4 to C8, in particular C6, alkylene chains, said cycloalkyl group in particular containing C4 to C12, in particular C7 to C9, notably C8, alkyl side groups.
In particular, the fatty acid dimer may be formed from C16 to C20, in particular C18 fatty acids, for example from oleic acid and/or linoleic acid.
Fatty acid dimers may be commercially available. As examples, mention may be made of the fatty acid dimers sold by the company Croda under the references Pripol®, notably Pripol®1009, of cosmetic grade, of structure (III) below:
in which formula (III) n is an integer between 6 and 8. According to a particular embodiment, the polyacid component b2) may comprise, or even consist of, diacid(s) of fatty acid dimer type, for example Pripol®1009.
The content of said polycarboxylic acid(s) b2), in particular of the polyacid fatty acid dimer type such as Pripol®1009, in b) according to the invention may be between 5% and 70% by weight, in particular between 15% and 65% by weight, more particularly from 25% to 55% by weight, notably from 25% to 50% by weight, relative to the total weight of the components forming b) according to the invention.
As described previously, b) may be obtained from at least one monocarboxylic acid component, also referred to hereinbelow as “monoacid”, denoted b3), i.e. comprising a single carboxyl group.
The monocarboxylic acids under consideration according to the invention are more particularly acyclic or (poly)cyclic, saturated or unsaturated, aromatic or unsaturated, linear or branched monocarboxylic acids comprising from 6 to 32 carbon atoms, in particular from 8 to 28 carbon atoms and more particularly from 10 to 20 carbon atoms.
It is understood that the oligo/polyester(s) b) may be prepared from a single monoacid or from a mixture of at least two monoacids. In the continuation of the text, unless otherwise specified, the term “monoacid” or “monocarboxylic acid” means a single monoacid compound or a mixture of several monoacid compounds.
According to a particular embodiment, said monocarboxylic acid b3) is non-aromatic, saturated or unsaturated, linear or branched, cyclic or non-cyclic, and comprises from 6 to 32 carbon atoms, in particular from 8 to 30 carbon atoms and more particularly from 10 to 28 carbon atoms, preferentially from 12 to 26 carbon atoms, more preferentially from 16 to 24 carbon atoms, even more preferentially from 18 to 22 carbon atoms.
Preferably, said monoacid component b3) is biobased, in particular of plant origin.
When the monoacid b3) used is of natural origin, it may notably consist of mixtures comprising saturated acids and unsaturated acids with conjugated and/or non-conjugated unsaturations.
According to a particular embodiment, said monocarboxylic acid b3) is a saturated or unsaturated, preferably biobased fatty acid.
According to a first particular embodiment, said monoacid b3) according to the invention is a monocarboxylic acid comprising a linear or branched, preferably linear, saturated or unsaturated aliphatic chain comprising more than 7 carbon atoms, in particular more than 8 carbon atoms, in particular from 8 to 30 carbon atoms, notably from 14 to 26 carbon atoms and more particularly from 16 to 22 carbon atoms, even more preferentially from 18 to 20 carbon atoms.
By way of example, a monoacid according to the invention may be chosen from isostearic acid (branched saturated C18), stearic acid (linear saturated C18), linoleic acid (linear polyunsaturated C18), arachidic acid (linear saturated C20), behenic acid (linear saturated C20), and behenic acid (linear saturated C22), capric acid (linear saturated C10), caprylic acid (linear saturated C8) and mixtures thereof, and also the geometrical isomers thereof, and the salts thereof with organic or mineral bases.
According to another particular embodiment, said monoacid b3) according to the invention is a (poly)cyclic, preferably polycyclic, monocarboxylic acid, in particular comprising a saturated and/or unsaturated, preferably unsaturated, in particular non-aromatic, monocyclic or fused bicyclic or tricyclic carbocycle, and including from 5 to 20 carbon atoms, said carbocycle being optionally substituted with one or more (C1-C4)alkyl groups such as methyl. In particular, said monoacid b3) according to the invention comprises a polycyclic, notably tricyclic, preferably unsaturated and non-aromatic, fused C8-C14 carbocycle, such as a partially hydrogenated phenanthrenyl group, in particular a decahydrophenanthrenyl group.
It may notably be abietic acid or an optical isomer thereof, a salt thereof with organic or mineral bases, and solvates thereof, such as hydrates. In particular, abietic acid may be derived from natural rosin.
It is understood that the particular embodiments described previously for each of the components b1), b2)) and b3) according to the invention may be combined.
According to one embodiment, the oligo/polyester(s) b) of the invention are derived from the condensation of polyol(s) b1) and monoacid(s) b3) such as a plant oil notably chosen from triglycerides.
According to one embodiment, the oligo/polyester(s) b) of the invention are derived from the condensation of polyol(s) b1) and of diacid(s) b2).
According to one embodiment, the oligo/polyester(s) b) of the invention are derived from the condensation of polyol(s) b1) and of diacid(s) b2) and of monoacid(s) b1).
Thus, according to a particular embodiment, the oligo/polyester(s) b) according to the invention may be formed at least, or even solely, from:
According to one embodiment, the oligo/polyester(s) b) according to the invention may be formed at least, or even solely, from:
According to a particular embodiment, said oligo/poly-ester(s) b) according to the invention may comprise, or even be formed of:
Advantageously, the oligo/polyester(s) b) used according to the invention are of biobased origin. In particular, it may be formed to more than 50% by weight, in particular to more than 80% by weight, or even exclusively formed, from biobased components (notably, polyacids, polyols and monoacids).
The oligo/polyester(s) b) used according to the invention may have an acid number of less than or equal to 5, in particular between 0.2 and 5 and more particularly between 0.5 and 3 mg KOH/g.
For the purposes of the invention, the term “acid number” means a number characterizing the acidity of the oligo/polyester(s) b). This number thus corresponds to the number of milligrams of potassium hydroxide required to neutralize one gram of sample of oligo/polyester(s) b). The acid number can be measured according to the ISO 2114 titration method.
The oligo/polyester(s) used according to the invention may have a weight average molar mass, Mw, ranging from 2000 to 10 000 g·mol−1, in particular from 3000 to 7000 g·mol−1.
The weight-average molar mass may be determined by size exclusion chromatography (SEC), as described more particularly in the following examples.
According to a particular embodiment of the invention the oligo/polyester(s) of the invention are liquid. According to a particular embodiment of the invention, the oligo/polyester(s) are liquid oligoesters.
The term “liquid” oligo/polyester means an oligo/polyester which begins to flow under its own weight in less than one minute at room temperature (25° C.) and at atmospheric pressure.
More particularly, the liquid oligo/polyester(s) are obtained by condensation of dimer(s) and/or trimer(s) of saturated or unsaturated fatty acids (preferably unsaturated fatty acid(s)) and of diol(s).
In the context of the present invention, the term “saturated fatty acids” means saturated fatty acids, i.e. fatty acids not comprising any unsaturation and comprising from 14 to 22 carbon atoms. Preferably, the saturated fatty acid dimers and/or trimers are polycarboxylic acids comprising at least 2 and up to 6 carboxylic acid groups per molecule. In a preferred embodiment, the saturated fatty acid dimer may comprise from 28 to 44 carbon atoms and 2 carboxylic acid groups. The saturated fatty acid trimer may comprise from 42 to 66 carbon atoms and 3 carboxylic acid functions.
Preferably, an unsaturated fatty acid dimer is used, in particular containing 36 carbon atoms and 2 carboxylic acid groups.
In the context of the present invention, the term “unsaturated fatty acids” means mono- or polyunsaturated fatty acids comprising from 14 to 22 carbon atoms. Unsaturated fatty acid dimers may notably comprise from 2 to 4 conjugated or unconjugated unsaturations, in their carbon chain. Unsaturated fatty acid trimers may comprise from 3 to 6 conjugated or unconjugated unsaturations, in their carbon chain. Preferably, the unsaturated fatty acid dimers and/or trimers are polycarboxylic acids comprising at least 2 and up to 6 carboxylic acid groups per molecule. In a preferred embodiment, the unsaturated fatty acid dimer may comprise from 28 to 44 carbon atoms and 2 carboxylic acid groups. The unsaturated fatty acid trimer may comprise from 42 to 66 carbon atoms and 3 carboxylic acid functions.
Preferably, an unsaturated fatty acid dimer is used, in particular containing 36 carbon atoms and 2 carboxylic acid groups.
Mixtures of unsaturated fatty acid dimers and trimers and/or unsaturated fatty acid (unpolymerized and therefore corresponding to a monomer) may also be used in the context of the invention. In the case of such a mixture, a mixture comprising more than 50% by weight of dimers is preferred, for example a mixture comprising more than 90% by weight, preferably more than 95%, of acids in the form of dimers, it being possible for the remainder of the mixture to be trimers and/or monomers of unsaturated fatty acids.
The unsaturated fatty acid dimer and/or trimer may optionally be hydrogenated after the polymerization reaction of the unsaturated fatty acid, notably to improve the stability of the dimer or trimer product.
Hydrogenated fatty acid dimers (oleic or linoleic acid) are notably sold under the brand names Empol1008, Empol1004, Empol1025, Empol1011 and Empol1062 by Cognis and Pripol 1006 (dilinoleic acid) by Uniqema, International. Uniqema also markets a hydrogenated fatty acid dimer under the name Pripol 1013 (hydrogenated dilinoleic acid).
Particularly preferably, the unsaturated fatty acid dimer is a dimer of linoleic acid, also called dilinoleic acid, obtained by intermolecular polymerization of linoleic acid.
The unsaturated fatty acid may be of natural origin, preferably of plant origin. A fatty acid of plant origin may be derived from any plant source producing said fatty acid. For example, in the case of linoleic acid, molecules extracted from soybean or oilseed rape may be used.
The oligo/polyester(s) in the composition according to the invention are thus obtained by condensation of a polymerized long-chain fatty acid with a diol. Preferably, the diol is a C2 to C10, preferably C2-C8, and preferentially C2-C6 hydrocarbon-based compound, the carbon chain of which is substituted with two hydroxyl functions. The hydrocarbon-based chain(s) may be interrupted with an oxygen atom. The diols that may be used according to the invention may be linear, branched or cyclic, saturated or unsaturated alcohols. Preferably, the diol is a linear saturated diol. Particularly preferably, the diol is a butanediol, notably 1,2-butanediol, 1,3-butanediol or 1,4-butanediol, and preferably 1,4-butanediol.
More particularly, the oligo/polyester(s) are b):
Advantageously, the oligo/polyester b) used in composition C1 according to the invention has an average molecular weight of between 500 and 10 000, preferably between 950 and 5500, and preferentially between 1200 and 1800.
According to one embodiment, the oligo/polyester(s) b) of the invention are chosen from copolymers of dinoleic acid and propanediol (INCI name: diniloleic acid propanediol copolymer, sold under the name Viscoplast Green 3000, INCI name: dilinoleic acid butanediol copolymer, sold under the name Viscoplast 14436 H, INCI name; dimer dinoleyl dimer dilinoleate, sold under the name Lusplan DD-DA7, INCI name: capryloyl glycerin/sebacic acid copolymer, sold under the name Lexfilm Sun Natural, INCI name: capryloyl glycerin/sebacic acid copolymer, sold under the name Vellaplex, INCI name: hydrogenated castor oil/sebacic acid copolymer, sold under the name Crodabond CSA, INCI name: pentaerytrithyl isostearate/caprate/adipate, sold under the name Supermol L-LQ, INCI name: diisostearyl polyglyceryl-3 dimer dilinoleate, sold under the name Solamaze Natural, INCI name: polyglyceryl-6 polyricenoleate, sold under the name S Face CR 1001, and INCI name: polyglyceryl-10 decaisostearate, sold under the name Nikkol Decaglyn 10 ISV.
In a particularly preferred embodiment, the polyester obtained by condensation of dimer and/or trimer of unsaturated fatty acid and of diol is a polymer, or polyester, of dilinoleic acid and of 1,4-butanediol, preferably having an average molecular weight of 1300, a viscosity at 40° C. of 2500-3500 cSt and a refractive index at 25° C. of 1.475-1.485. Mention may notably be made in this respect of the polymer sold by Biosynthis under the name Viscoplast 14436H (INCI name: dilinoleic acid/butanediol copolymer).
Preferably, the lower limit of the viscosity of the oligo/polyester(s) b) of the invention is 800 mPas at 25° C.
Preferably, the oligo/polyester(s) b) of the invention are chosen from i) pentaerythrityl isostearate/caprate/caprylate/adipate; and ii) dimer dilinoleyl dimer dilinoleate, more preferentially ii).
The total amount of the polyester(s) or oligoester(s) b), present in composition C1 according to the invention, is preferably from 0.01% to 30% by weight, more preferentially from 0.5% to 20% by weight, and better still from 1% to 15% by weight, such as 6% or 10% by weight relative to the total weight of the composition. According to a preferred embodiment of the invention, the total amount of the polyester(s) or oligoester(s) b), present in composition C1 according to the invention, is from 0.01% to 15% by weight, even better from 1% to 15% by weight, relative to the total weight of the composition.
The weight ratio between the total amount of a) PHA (active material) and the total amount b) of oligo/polyester(s) b), present in the composition according to the invention, is preferably from 0.5 to 10, more preferentially from 1 to 6, more particularly from 1 to 3, or even 1 to 2 such as 1.
c) The fatty substances
According to a particular embodiment of the invention, the composition also comprises one or more fatty substances.
The term “fatty substance” means an organic compound that is insoluble in water at ordinary room temperature (25° C.) and at atmospheric pressure (760 mmHg) (solubility of less than 5%, preferably 1% and even more preferentially 0.1%). They bear in their structure at least one hydrocarbon-based chain including at least 6 carbon atoms or a sequence of at least two siloxane groups. In addition, the fatty substances are generally soluble in organic solvents under the same temperature and pressure conditions, for instance chloroform, ethanol, benzene, liquid petroleum jelly or decamethylcyclopentasiloxane.
The fatty substance(s) of the invention are of natural or synthetic origin, preferably natural, more preferentially of plant origin. They are different from fatty acids since salified fatty acids constitute soaps which are generally soluble in aqueous media.
According to a particular embodiment of the invention, the composition comprises one or more fatty substances that are not liquid at 25° C. and at atmospheric pressure.
According to a particular embodiment, the composition of the invention comprises one or more waxes.
The term “wax” means a lipophilic compound that is solid at room temperature (25° C.), with a reversible solid/liquid change of state, having a melting point of greater than or equal to 30° C., which may be up to 200° C. and notably up to 120° C.
In particular, the wax(es) that are suitable for use in the invention may have a melting point of greater than or equal to 45° C. and in particular of greater than or equal to 55° C.
According to a particular form of the invention, the composition of the invention is solid, in particular anhydrous. It may then be in stick form; use will be made of polyethylene microwaxes in the form of crystallites with an aspect ratio at least equal to 2, and with a melting point ranging from 70 to 110° C. and preferably from 70 to 100° C., so as to reduce or even eliminate the presence of strata in the solid composition. These crystallites in needle form and notably the dimensions thereof may be characterized visually according to the following method.
According to a particular embodiment, the composition of the invention comprises one or more pasty compounds.
For the purposes of the present invention, the term “pasty compound” means a lipophilic fatty compound that undergoes a reversible solid/liquid change of state, having anisotropic crystal organization in the solid state, and including, at a temperature of 23° C., a liquid fraction and a solid fraction.
Preferably, the composition contains one or more fatty substances c) which are hydrocarbon-based fatty substances that are liquid at 25° C. and atmospheric pressure.
The hydrocarbon-based liquid fatty substance(s) are notably chosen from C6-C16 hydrocarbons or hydrocarbons comprising more than 16 carbon atoms and up to 60 carbon atoms, preferably between C6 and C16, and in particular alkanes, oils of animal origin, oils of plant origin, glycerides or fluoro oils of synthetic origin, fatty alcohols, fatty acid and/or fatty alcohol esters, and silicones. In particular, the liquid fatty substance(s) are chosen from non-silicone oils.
It is recalled that, for the purposes of the invention, the fatty alcohols, fatty esters and fatty acids more particularly contain one or more linear or branched, saturated or unsaturated hydrocarbon-based groups comprising 6 to 60 carbon atoms, which are optionally substituted, in particular with one or more hydroxyl groups OH (in particular from 1 to 4 hydroxyl groups). If they are unsaturated, these compounds may comprise one to three unsaturations, preferably from one to three conjugated or unconjugated carbon-carbon double bonds.
As regards the C6-C16 alkanes, these compounds are linear or branched, and optionally cyclic; preferably, the fatty substance(s) c) of the invention are chosen from linear or branched C8-C14, more preferentially C9-C13 and even more preferentially C9-C12 alkanes. Examples that may be mentioned include hexane, decane, undecane, dodecane, tridecane, and isoparaffins, for instance isohexadecane, isodecane or isododecane. The linear or branched hydrocarbons containing more than 16 carbon atoms may be chosen from liquid paraffins, liquid petroleum jelly, polydecenes, and hydrogenated polyisobutene such as Parleam®.
Among the hydrocarbon-based liquid fatty substances c) having an overall solubility parameter according to the Hansen solubility space of less than or equal to 20 (MPa)1/2, mention may be made of oils, which may be chosen from natural or synthetic, hydrocarbon-based oils, which are optionally fluorinated and optionally branched, alone or as a mixture.
According to a very advantageous embodiment, the composition of the invention comprises one or more fatty substances which are one or more hydrocarbon-based oils. The hydrocarbon-based oil(s) may be volatile or non-volatile.
According to a preferred embodiment of the invention, the fatty substance(s) c) are linear or branched hydrocarbon-based oils, which are volatile, notably chosen from undecane, decane, dodecane, isododecane, tridecane, and a mixture of various volatile oils thereof preferably comprising isododecane in the mixture, or a mixture of undecane and tridecane.
According to another particular embodiment, the liquid fatty substance(s) c) are a mixture of a volatile hydrocarbon-based oil and a non-volatile hydrocarbon-based oil, the mixture of which preferentially comprises dodecane or isododecane as volatile oil.
In particular, the fatty substance(s) c) of the invention are a mixture of C9-C12 alkanes, preferably of natural origin, the chains of which comprise from 9 to 12 carbon atoms, preferably linear or branched C9-C12 alkanes. This mixture is notably known under the INCI name C9-C12 Alkane, CAS 68608-12-8, Vegelight Silk® sold by BioSynthls. This volatile biodegradable mixture of volatile oils is obtained from coconut oil (the viscosity is 0.9-1.1 cSt (40° C.) and it has a flash point of 65° C.).
According to one embodiment, composition C1 contains only oils that are liquid at 25° C. and atmospheric pressure. According to another embodiment, composition C1 contains at least 80% of hydrocarbon-based oils that are liquid at 25° C. and atmospheric pressure, which are preferably volatile, more preferentially chosen from isodecane, decane, Cetiol UT® and Vegelight Silk®.
According to another embodiment, composition C1 may comprise volatile and non-volatile oils, notably in a volatile oil/non-volatile oil ratio of greater than or equal to 4.
According to another embodiment, composition C1 contains from 0 to 10% of silicone oils, preferably from 0 to 5% of silicone oils.
Volatile silicone oils that may be mentioned include volatile linear or cyclic silicone oils, notably those with a viscosity of less than or equal to 8 centistokes (cSt) (8×10−6 m2/s), and notably containing from 2 to 10 silicon atoms and in particular from 2 to 7 silicon atoms, these silicones optionally including alkyl or alkoxy groups containing from 1 to 10 carbon atoms. As volatile silicone oils that may be used in the invention, mention may notably be made of dimethicones with viscosities of 5 and 6 cSt, octamethylcyclotetrasiloxane, decamethylcyclopentasiloxane, dodecamethylcyclohexasiloxane, heptamethylhexyltrisiloxane, heptamethyloctyltrisiloxane, hexamethyldisiloxane, octamethyltrisiloxane, decamethyltetrasiloxane and dodecamethylpentasiloxane, and mixtures thereof.
As nonvolatile silicone oils, mention may be made of linear or cyclic nonvolatile polydimethylsiloxanes (PDMSs); polydimethylsiloxanes including alkyl, alkoxy and/or phenyl groups, which are pendent or at the end of a silicone chain, these groups containing from 2 to 24 carbon atoms; phenyl silicones, for instance phenyl trimethicones, phenyl dimethicones, phenyltrimethylsiloxydiphenylsiloxanes, diphenyl dimethicones, diphenylmethyldiphenyltrisiloxanes, 2-phenylethyl trimethylsiloxysilicates and pentaphenyl silicone oils.
The hydrocarbon-based oil may be chosen from:
In particular, the composition comprises at least one hydrocarbon-based liquid fatty substance c) chosen from:
Preferably, the composition of the invention comprises at least one hydrocarbon-based liquid fatty substance c) chosen from:
Advantageously, the fatty substance(s) c) of the invention, which are notably liquid, are apolar, i.e. formed solely of carbon and hydrogen atoms.
The hydrocarbon-based liquid fatty substance(s) are preferably chosen from hydrocarbon-based oils containing from 8 to 14 carbon atoms, which are in particular volatile, more particularly the apolar oils described previously.
Preferentially, the fatty substance(s) c) of the invention, which are notably liquid, are chosen from alkanes such as dodecane, decane, undecane, tridecane, isododecane, hydrogenated polyisobutene, fatty alcohols such as octyldodecanol, esters such as isononyl isononanoate, cocoyl caprylate/caprate and mixtures thereof, more preferentially alkanes.
More particularly, the fatty substance(s) c) of the invention, which are notably liquid, are chosen from linear or branched C6-C16, preferably C8-C14, more preferentially C9-C13 and even more preferentially C9-C12 alkanes, and even more preferentially the alkanes are volatile. More particularly, the liquid fatty substance(s) c) of the invention are volatile and are chosen from undecane, decane, dodecane, isododecane, tridecane, and a mixture thereof notably comprising dodecane, isododecane or a mixture of undecane and tridecane.
Preferentially, the liquid fatty substance(s) c) of the invention, which are notably liquid, are isododecane.
According to another advantageous embodiment of the invention, the fatty substance(s) c) of the invention, which are notably liquid, are a mixture of non-volatile oil(s) and volatile oil(s); preferably, the mixture comprises, as volatile oil, undecane, dodecane, isododecane or tridecane, more preferentially isododecane. A mixture of volatile oil and non-volatile oil that may be mentioned is the mixture of isododecane and of isononyl isononanoate or the mixture of isododecane with isononyl isononanoate.
More preferentially, when the fatty substance(s) are a mixture of volatile oil and of non-volatile oil, the amount of volatile oil is greater than the amount of non-volatile oil.
In particular, in the mixture, the non-volatile oil is a phenyl silicone oil, preferably chosen from pentaphenyl silicone oils.
Advantageously, composition C1 comprises one or more fatty substances, which are notably liquid at 25° C. and at atmospheric pressure, preferably one or more oils, in a content ranging from 2% to 99.9% by weight, relative to the total weight of the composition, preferably ranging from 5% to 90% by weight, preferably ranging from 10% to 80% by weight, preferably ranging from 20% to 80% by weight.
According to a preferred embodiment of the invention, composition C1 according to the invention comprises c) one or more fatty substances that are notably liquid at 25° C. and at atmospheric pressure, f) one or more organic solvents other than c) and optionally e) water.
d) Organic Solvent(s) Other than c)
Preferably, composition C1 also comprises one or more organic solvents other than c), which are apolar or polar, preferably polar, and which are protic or aprotic, more particularly protic and/or polar, preferably protic and polar.
Preferably, the organic solvent(s) are water-miscible.
According to the invention, the term “water-miscible solvent” is intended to denote a compound that is liquid at room temperature and water-miscible (water miscibility of greater than 50% by weight at 25° C. and atmospheric pressure).
The organic solvent(s) that may be used in composition C1 of the invention may also be volatile.
Among the organic solvents that may be used in composition C1 according to the invention, mention may notably be made of polar protic or polar aprotic organic solvents, preferably polar protic organic solvents, particularly lower monoalcohols containing from 2 to 10 carbon atoms, such as ethanol and isopropanol, preferably ethanol.
According to one embodiment, composition C1 of the invention comprises one or more organic solvents, preferably chosen from monoalcohols containing from 2 to 6 carbon atoms such as ethanol and isopropanol.
Preferably, the composition according to the invention also comprises at least one polar organic solvent other than the fatty substances c), more preferentially protic solvent.
More preferentially, the composition according to the invention also comprises at least one polar organic solvent other than the fatty substances c), chosen from lower monoalcohols containing from 2 to 10 carbon atoms, such as ethanol and isopropanol, and even more preferentially ethanol.
Preferably, the amount of organic solvent(s) is less than 70% by weight, more preferentially less than 50% by weight, relative to the total weight of composition C1. According to one embodiment of the invention, composition C1 comprises an amount of organic solvent(s) of greater than 0.1%, more particularly greater than or equal to 0.5% by weight relative to the total weight of the composition. In particular, the composition comprises between 1% and 50% by weight of organic solvent(s), more particularly between 2% and 10% and better still between 2.5% and 5%.
Preferably, the content of polar organic solvent(s) other than the fatty substances c) present in the composition according to the invention is in the range from 1% to 50% by weight, more preferentially from 1% to 30% by weight, even more preferentially from 1% to 20% by weight, better still from 1% to 10% by weight, even better still from 2% to 10% by weight, or even from 2% to 5% by weight and even better still from 2.5% to 5% by weight, relative to the total weight of the composition according to the invention.
Preferably, the content of polar protic organic solvent(s) other than the fatty substances c) present in the composition according to the invention is in the range from 1% to 50% by weight, more preferentially from 1% to 30% by weight, even more preferentially from 1% to 20% by weight, better still from 1% to 10% by weight, even better still from 2% to 10% by weight, or even from 2% to 5% by weight and even better still from 2.5% to 5% by weight, relative to the total weight of the composition according to the invention.
Preferably, the content of lower monoalcohols containing from 2 to 10 carbon atoms present in the composition according to the invention is in the range from 1% to 50% by weight, more preferentially from 1% to 30% by weight, even more preferentially from 1% to 20% by weight, better still from 1% to 10% by weight, even better still from 2% to 10% by weight, or even from 2% to 5% by weight and even better still from 2.5% to 5% by weight, relative to the total weight of the composition according to the invention.
Preferably, the content of ethanol in the composition according to the invention is in the range from 1% to 50% by weight, more preferentially from 1% to 30% by weight, even more preferentially from 1% to 20% by weight, better still from 1% to 10% by weight, even better still from 2% to 10% by weight, or even from 2% to 5% by weight and even better still from 2.5% to 5% by weight, relative to the total weight of the composition according to the invention.
According to a particular embodiment of the invention, composition C1 also comprises water.
The water that is suitable for use in the invention may be tap water, distilled water, spring water, a floral water such as cornflower water and/or a mineral water such as Vittel water, Lucas water or La Roche Posay water and/or a thermal water.
According to one embodiment, composition C1 of the invention comprises e) water and at least one fatty substance c) in a ratio between the mass of water and the mass of fatty substance c) of less than 1, preferably less than 0.9, more preferentially less than 0.9, such as between 0.5 and 0.8.
According to a particular embodiment of the invention, composition C1 comprises an amount of water of less than or equal to 10% by weight relative to the total weight of the composition, particularly less than or equal to 5% by weight, preferably less than 2% by weight, more preferentially less than 1% by weight relative to the total weight of the composition. More particularly, the composition of the invention is anhydrous, i.e. it is free of water.
According to a particular embodiment of the invention, composition C1 also comprises f) one or more surfactants, preferably nonionic or ionic surfactants, or mixtures thereof.
According to another particular embodiment of the invention, composition C1 does not comprise any surfactant.
The term “surfactant” means a compound which modifies the surface tension between two surfaces. The surfactant(s) d) are amphiphilic molecules, which have two parts of different polarity, one part being lipophilic (which retains fatty substances) which is apolar, the other hydrophilic part (miscible or soluble in water) being polar. The lipophilic part is generally a fatty chain, and the other water-miscible part is polar, and/or protic.
The term “ionic” means anionic, cationic, amphoteric or zwitterionic.
The term “fatty chain” means a linear or branched, saturated or unsaturated hydrocarbon-based chain comprising more than 6 atoms, preferably between 6 and 30 carbon atoms and preferably from 8 to 24 carbon atoms.
According to a first particular embodiment, the composition of the invention contains d) at least one silicone or non-silicone nonionic surfactant.
Among the nonionic surfactants according to the invention, mention may be made, alone or as mixtures, of fatty alcohols, α-diols and alkylphenols, these three types of compound being polyethoxylated, polypropoxylated and/or polyglycerolated and containing a fatty chain comprising, for example, 8 to 22 carbon atoms, the number of ethylene oxide or propylene oxide groups possibly ranging in particular from 2 to 50 and the number of glycerol groups possibly ranging in particular from 2 to 30. Mention may also be made of ethylene oxide and propylene oxide copolymers, condensates of ethylene oxide and of propylene oxide with fatty alcohols; polyethoxylated fatty amides preferably having from 2 to 30 ethylene oxide units, polyglycerolated fatty amides containing on average 1 to 5, and in particular 1.5 to 4, glycerol groups, ethoxylated fatty acid esters of sorbitan containing from 2 to 30 ethylene oxide units, fatty acid esters of sucrose, fatty acid esters of polyethylene glycol, alkylpolyglycosides, N-alkylglucamine derivatives, amine oxides such as (C10-C14)alkylamine oxides or N-acylaminopropylmorpholine oxides.
The surfactant(s) represent in total particularly from 0.01% to 30% by weight, preferably from 0.5% to 15% by weight, even more preferentially from 1% to 10% by weight and better still between 1% and 5% by weight of the composition, relative to the total weight of the composition.
According to one embodiment of the invention, composition C1 comprises an aqueous phase. The composition is notably formulated as aqueous lotions or as water-in-oil or oil-in-water emulsions or as multiple emulsions (oil-in-water-in-oil or water-in-oil-in-water triple emulsions (such emulsions are known and described, for example, by C. Fox in “Cosmetics and Toiletries”—November 1986—Vol. 101—pages 101-112)).
According to a particular embodiment of the invention, the composition is a direct emulsion, i.e. an emulsion of oil-in-water or O/W type. The weight amount of oil is preferably less than 70% in the inverse emulsion, preferably less than or equal to 40%, more particularly less than or equal to 35% by weight relative to the total weight of the composition.
More particularly, in the direct emulsion, the amount of water is greater than or equal to 30% by weight relative to the total weight of the composition, more particularly greater than or equal to 40%, preferentially greater than or equal to 35%.
According to another particular embodiment of the invention, the composition of the invention is an inverse emulsion, i.e. of water-in-oil or W/O type. The weight amount of oil is preferably greater than 30% in the inverse emulsion, preferably greater than 40%, more preferentially greater than or equal to 45% by weight relative to the total weight of the composition. More particularly, in the inverse emulsion, the amount of water is less than 40% by weight relative to the total weight of the composition, preferably less than or equal to 35% by weight.
According to one embodiment, the composition is an oily composition comprising at least one fatty substance c) and optionally at least one solvent d), preferably containing at least one fatty substance c) and at least one solvent d).
Composition C1 according to the invention preferably has a pH ranging from 3 to 9, depending on the support chosen.
According to a particular embodiment of the invention, the pH of the composition(s) is neutral or even slightly acidic. Preferably, the pH of the composition is between 6 and 7. The pH of these compositions may be adjusted to the desired value by means of acidifying or basifying agents usually used in cosmetics, or alternatively using standard buffer systems.
The term “basifying agent” or “base” means any agent for increasing the pH of the composition in which it is present. The basifying agent is a Brønsted, Lowry or Lewis base. It may be mineral or organic. Particularly, said agent is chosen from a) aqueous ammonia, b) (bi)carbonate, c) alkanolamines such as monoethanolamine, diethanolamine, triethanolamine and derivatives thereof, d) oxyethylenated and/or oxypropylenated ethylenediamines, e) organic amines, f) mineral or organic hydroxides, g) alkali metal silicates such as sodium metasilicates, h) amino acids, preferably basic amino acids such as arginine, lysine, ornithine, citrulline and histidine, and i) the compounds of formula (F) below:
in which formula (F):
Examples of amines of formula (F) that may be mentioned include 1,3-diaminopropane, 1,3-diamino-2-propanol, spermine and spermidine.
The term “alkanolamine” means an organic amine comprising a primary, secondary or tertiary amine function, and one or more linear or branched C1-C8 alkyl groups bearing one or more hydroxyl radicals.
Among the mineral or organic hydroxides, mention may be made of those chosen from a) hydroxides of an alkali metal, b) hydroxides of an alkaline-earth metal, for instance sodium hydroxide or potassium hydroxide, c) hydroxides of a transition metal, d) hydroxides of lanthanides or actinides, quaternary ammonium hydroxides and guanidinium hydroxide. The mineral or organic hydroxides a) and b) are preferred.
Mention may be made, among the acidifying agents for the compositions used in the invention, by way of example, of mineral or organic acids, such as hydrochloric acid, orthophosphoric acid, sulfuric acid, carboxylic acids, such as acetic acid, tartaric acid, citric acid or lactic acid, or sulfonic acids.
The basifying agents and the acidifying agents as defined previously preferably represent from 0.001% to 20% by weight relative to the weight of the composition. and more particularly from 0.005% to 8% by weight of the composition.
According to a particular embodiment of the invention, composition C1 comprises an amount of water of less than or equal to 10% by weight relative to the total weight of the composition. Even more preferentially, composition C1 comprises an amount of water of less than or equal to 5%, better still less than 2%, even better still less than 0.5%, and is notably free of water. Where appropriate, such small amounts of water may notably be introduced by ingredients of the composition that may contain residual amounts thereof.
According to a particular embodiment of the invention, the composition does not comprise any water.
Advantageously, composition C1 according to the invention comprises a physiologically acceptable medium. In particular, the composition is a cosmetic composition.
The term “physiologically acceptable medium” means a medium that is compatible with human keratin materials, for instance the skin, the lips, the nails, the eyelashes, the eyebrows or the hair.
The term “cosmetic composition” means a composition that is compatible with keratin materials, which has a pleasant colour, odour and feel and which does not cause any unacceptable discomfort (stinging or tautness) liable to discourage the consumer from using it.
The term “keratin materials” means the skin (body, face, contour of the eyes, scalp), head hair, the eyelashes, the eyebrows, bodily hair, the nails or the lips.
Composition C1 according to the invention may comprise one or more cosmetic additives chosen from fragrances, preserving agents, fillers, colouring agents, UV-screening agents, oils other than the fatty substances c), moisturizers, vitamins, ceramides, antioxidants, free-radical scavengers, polymers other than a), thickeners or film-forming agents other than b), trace elements, softeners, sequestrants, agents for combating hair loss, anti-dandruff agents, propellants. In particular, composition C1 according to the invention also comprises one or more colouring agents chosen from pigments, direct dyes and mixtures thereof, preferably pigments; more preferentially, the pigment(s) of the invention are chosen from carbon black, iron oxides, notably black iron oxides, and micas coated with iron oxide, triarylmethane pigments, notably blue and violet triarylmethane pigments, such as Blue 1 Lake, azo pigments, notably red azo pigments, such as D&C Red 7, an alkali metal salt of lithol red, such as the calcium salt of lithol red B, even more preferentially red iron oxides.
Advantageously, composition C1 according to the invention is a makeup composition, in particular a lip makeup composition, a mascara, an eyeliner, an eye shadow or a foundation.
Composition C1 according to the invention may also comprise one or more fillers, notably in a content ranging from 0.01% to 30% by weight and preferably ranging from 0.01% to 20% by weight relative to the total weight of the composition. The term “fillers” should be understood as meaning colourless or white, mineral or synthetic particles of any shape, which are insoluble in the medium of the composition, irrespective of the temperature at which the composition is manufactured. These fillers notably serve to modify the rheology or texture of the composition.
Composition C1 according to the invention may be in the form of an anhydrous composition, a water-in-oil emulsion or an oil-in-water emulsion.
The invention is illustrated in greater detail in the examples that follow. The amounts are indicated as weight percentages.
The PHAs illustrated in the various examples were prepared in 3-litre chemostats and/or 5-litre Fernbach flasks depending on whether or not a β-oxidation pathway inhibitor was used. The isolation of the PHAs is similar for all the examples obtained.
In a first step, the microorganism generates the PHAs which are stored in intracellular granules, the proportion of which varies as a function of the applied conditions such as the temperature or the nature of the culture medium. The generation of PHA granules may or may not be associated with the growth of the microorganism as a function of the nature of the microorganisms. During the second step, the biomass containing the PHAs is isolated, i.e. separated from the fermentation medium, and then dried. The PHAs are extracted from the biomass before being purified, if necessary.
A mixture of saturated and unsaturated carbon sources is, for certain examples, necessary for the stability of the PHA obtained.
The process for synthesizing the compound of Example 1 is adapted from the article: Fed-batch production of unsaturated medium-chain-length polyhydroxyalkanoates with controlled composition by Pseudomonas putida KT2440, Z. Sun, J. A. Ramsay, M. Guay, B. A. Ramsay, Applied Microbiology Biotechnology, 82. 657-662, 2009.
The microorganism used is Pseudomonas putida KT2440 ATCC® 47054™. The culture method is performed under fed-batch growth axenic conditions with a maintenance solution containing a mixture of carbon source at a rate μ=0.15 h−1 in a 3 L chemostat containing 2.5 L of culture medium.
The system is aerated with a flow of 0.5 vvm of air for a nominal dissolved oxygen (OD) value at 30% of saturation. The pH is regulated with 15% aqueous ammonia solution. The temperature of the fermentation medium is regulated at 30° C.
Equipment for the fed-batch growth fermentation mode:
The composition of the Nutrient Broth, as mass percentages, is 37.5% beef extract and 62.5% peptone. Reference 233000 DIFCO™.
100 mL of preculture are prepared by suspending a cryotube containing 1 mL of the strain with 100 mL of “inoculum” culture medium at a pH adjusted to 6.8 with 2N NaOH in a 250 mL Fernbach flask and are then incubated at 30° C. at 150 rpm for 24 hours. 1.9 L of CM2 “batch” culture medium placed in a presterilized 3 L chemostat are inoculated at OD=0.1 with the 100 mL of preculture. After 4 hours at 30° C. at 850 rpm.
At the end of the introduction, the biomass is isolated by centrifugation and then washed three times with water. The biomass is dried by lyophilization before being extracted with ethyl acetate for 24 hours. The suspension is clarified by filtration on a GF/A filter (Whatman®). The filtrate, composed of PHA dissolved in the ethyl acetate, is concentrated by evaporation and then dried under high vacuum at 40° C. to constant mass.
The PHA may optionally be purified by successive dissolution and precipitation from an ethyl acetate/ethanol 70% methanol system, for example.
The PHA was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure.
Preparation of Example 1′: PHA Copolymer Bearing a Side Chain R1 Representing a 5% Unsaturated n-Octenyl Group and R2 Representing an n-Hexyl Group
The copolymer of Example 1′ (5% unsaturation and R2 chain representing n-hexyl) was prepared according to the procedure described for Example 1, with the same composition of the microelement solution as described in Example 1 and with the following culture medium compositions:
The PHA copolymer of Example 1′ was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure, with a degree of unsaturation of 5%.
The copolymer of Example 1″ (10% unsaturation and R2 chain representing n-hexyl) was prepared according to the procedure described for Example 1, with the same composition of the microelement solution as described in Example 1 and with the following culture medium compositions:
The PHA was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure.
The copolymer of Example 1′″ (30% unsaturated and R2 chain representing n-pentyl) was prepared according to the procedure described for Example 1, with the same composition of the microelement solution as described in Example 1 and with the following culture medium compositions:
The PHA copolymer was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure.
The process for obtaining example 1d is adapted from Appl Microbiol Biotechnol 82:657-662 (2009).
“Fed-batch production of unsaturated medium-chain-length polyhydroxyalkanoates with controlled composition by Pseudomonas putida KT2440”.
The microorganism used is Pseudomonas putida KT2440 ATCC®47054™.
The culture mode is carried out under axenic conditions in discontinuous growth fed with a maintenance solution containing a mixture of carbon sources at a rate of μ=0.15 h−1 in a 3 L chemostat containing 2.5 L of medium of culture. The flow rate of the maintenance supply pump is proportional to the growth of the microorganism according to formula 1:
Formula 1: theoretical equation linking the quantity of biomass and carbon source as a function of time with St=quantity of carbon source required to produce the biomass Xt at time t (g), YX/S=biomass yield from the carbon source, X0=initial biomass (g) and μ=desired specific growth rate (h−1).
The system is aerated by an air flow of 0.5 vvm for a dissolved oxygen (DO) setpoint at 30% saturation. The pH is regulated with a 15% of ammonia solution. The temperature of the fermentation medium is regulated at 30° C. The Assembly of the fed batch growth fermentation mode is made according
The fermentation medium is regulated in temperature-dissolved oxygen pressure and pH (not shown on the fig.).
The production process is carried out using three distinct culture media. The first culture medium defined MC1 “inoculum” is used for the preparation of the preculture.
The second culture medium defined MC2 “bach” is used for the non-supplied discontinuous growth of the microorganism with the primary carbonaceous sources in the Fernbachs flasks.
The third culture medium defined (MC3 “maintenance”) is used for the discontinuous feeding, or maintenance, of the fermentation with the carbonaceous sources of interest at a rate calibrated according to the growth of the microorganism.
The composition in grams per liter of the three media is described in Table below:
The composition of Nutrient Broth in mass percentage is 37.5% beef extract and 62.5% peptone. Reference 233000 DIFCO™.
The composition of the solution of microelements in grams per liter is described in Table below:
100 mL of preculture are prepared by suspending a cryotube containing 1 mL of the strain with 100 mL “inoculum” culture media at pH adjusted to 6.8 with 2N NaOH in a 250 mL Fernbach flask then incubating at 30° C. at 150 rpm for 24 hours. 1.9 L of MC2 “BATCH” culture medium placed in a previously sterilized 3 L chemostat are inoculated at OD=0.1 with the 100 mL of preculture. After 4 hours at 30° C. at 850 rpm, the introduction of the maintenance is carried out by applying the flow rate defined by equation 1. At the end of the introduction, the biomass is isolated by centrifugation then washed three times with some water. The biomass is dried by freeze-drying before being extracted with dichloromethane for 24 hours. The suspension is clarified by filtration on a GF/A filter (Wattman®) the filtrate, composed of PHA in solution in dichloromethane, is concentrated by evaporation then dried under high vacuum at 40° C. until constant mass. The PHA can optionally be purified by solubilization and successive precipitations such as a dichloromethane methanol system for example.
The PHA was characterized by gas chromatography equipped with an FID detector. It conforms to the expected chemical structure, with an unsaturation rate of 2%.
1 g of the compound of Example 1 and 150 mg of thiolactic acid were dissolved in 20 mL of ethyl acetate at room temperature with stirring. 20 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.
20 mL of the reaction medium were then precipitated from a 200 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.
The grafted PHA of Example 2 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure.
0.5 g of the compound of Example 1 and 125 mg of octanethiol were dissolved in 10 mL of ethyl acetate at room temperature with stirring. 15 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.
The reaction medium was then precipitated from a 100 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.
The grafted PHA of Example 3 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure.
50 mg of the compound of Example 1 and 10 mg of 8-mercapto-1-octanol were dissolved in 5 mL of ethyl acetate at room temperature with stirring. 2 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.
The reaction medium was then precipitated from a 50 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.
The grafted PHA of Example 4 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 75% or 7.5% of functions in total.
0.5 g of the compound of Example 1 and 54 mg of cysteamine were dissolved in a mixture of 10 mL of dichloromethane and 2 mL of ethanol at room temperature with stirring. 10 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.
The reaction medium was then precipitated from a 100 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.
The grafted PHA of Example 5 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 32% or 3.2% of functions in total.
100 mg of the compound of Example 1 and 26 mg of cyclohexanethiol were dissolved in 5 mL of dichloromethane at room temperature with stirring. 5 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.
The reaction medium was then precipitated from a 50 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.
The grafted PHA of Example 6 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 73% or 7.3% of functions in total.
100 mg of the compound of Example 1 and 26 mg of 2-furanmethanethiol were dissolved in 5 mL of dichloromethane at room temperature with stirring. 5 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.
The reaction medium was then precipitated from a 50 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.
The grafted PHA of Example 7 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 70% or 7% of functions in total.
100 mg of the compound of Example 1 and 26 mg of 1-thio-β-D-glucose tetraacetate were dissolved in 5 mL of dichloromethane at room temperature with stirring. 5 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.
The reaction medium was then precipitated from a 50 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.
The grafted PHA of Example 8 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 70% or 7% of functions in total.
100 mg of the compound of Example 1 and 26 mg of 2-phenylethanethiol were dissolved in 5 mL of dichloromethane at room temperature with stirring. 5 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.
The reaction medium was then precipitated from a 50 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.
The grafted PHA of Example 9 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 50% or 5% of functions in total.
100 mg of the compound of Example 1 and 26 mg of 4-tert-butylbenzyl mercaptan were dissolved in 5 mL of dichloromethane at room temperature with stirring. 5 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.
The reaction medium was then precipitated from a 50 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.
The grafted PHA of Example 10 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 64% or 6.4% of functions in total.
0.1 g of the compound of Example 1″ and 15 mg of thiolactic acid were dissolved in 5 mL of chloroform at room temperature with stirring. 5 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.
The reaction medium was then precipitated from a 50 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.
The grafted PHA of Example 11 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 100%.
2 g of compound of example 1d and 180 mg of thiolactic acid were dissolved in 15 mL of ethyl acetate at room temperature with stirring. 5 mg of 2-Hydroxy-2-methylpropiophenone (HMP) was added to the mixture.
The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes. The reaction medium thus obtained is poured onto a Teflon plate, then dried under dynamic vacuum at 40° C., to obtain a homogeneous film. The PHA grafted with thiolactic acid was fully characterized by proton NMR. The proton NMR spectrum shows that the characteristic signals of the unsaturations have completely disappeared.
1 g of the PHA copolymer of Example 1′ and 150 mg of octanethiol were dissolved in 15 mL of ethyl acetate at room temperature with stirring. 20 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.
The reaction medium was then precipitated from a 500 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.
The grafted PHA of Example 12 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 100%.
20 g of the PHA copolymer of Example 1′ were dissolved in 80 mL of anhydrous dichloromethane. A suspension of 1.9 g of 77% m-CPBA was prepared with 20 mL of anhydrous dichloromethane and added to the mixture with stirring, at room temperature for at least 120 hours.
The reaction medium was then precipitated from a 500 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.
The PHA of Example 13 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Epoxidation to 100%.
10 g of the PHA copolymer of Example 1″ were dissolved in 40 mL of anhydrous dichloromethane. A suspension of 1.9 g of 77% m-CPBA was prepared with 10 mL of anhydrous dichloromethane and added to the mixture with stirring, at room temperature for at least 120 hours.
The reaction medium was then precipitated from a 500 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.
The PHA of Example 14 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Epoxidation to 100%.
10 g of the PHA copolymer of Example 1′″ were dissolved in 40 mL of anhydrous dichloromethane. A suspension of 6.2 g of 77% m-CPBA was prepared with 10 mL of anhydrous dichloromethane and added to the mixture with stirring, at room temperature for at least 120 hours.
The reaction medium was then precipitated from a 250 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.
The PHA of Example 15 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Epoxidation to 100%.
2 g of the PHA copolymer of Example 1′ and 300 mg of 4-tert-butylbenzyl mercaptan were dissolved in 25 mL of ethyl acetate at room temperature with stirring. 25 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.
The reaction medium was then precipitated from a 500 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.
The PHA of Example 16 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 100%.
The production process of Example 17 is an adaptation of Applied and Environmental Microbiology, Vol. 60, No. 9. 3245-3254 (1994) “Polyester Biosynthesis Characteristics of Pseudomonas citronellolis Grown on Various Carbon Sources, Including 3-Methyl-Branched Substrate”. Mun Hwan Choi and Sung Chul Yoon. The microorganism used is Pseudomonas citronellolis ATCC® 13674™. The culture method was performed under axenic conditions in unfed batch culture mode in 5 L Fernbach flasks (Corning® ref. 431685) containing 2 of culture medium, shaken at 110 rpm at 30° C. in an orbital incubator (orbit diameter of 2.5 cm).
The production process is performed using two different culture media. The first culture medium, defined CM1 “inoculum”, is used for the preparation of the preculture. The second culture medium, defined CM2 “batch”, is used for unfed batch culture growth of the microorganism with the carbon source of interest in the Fernbach flasks.
The composition of the Nutrient Broth, as mass percentages, is 37.5% beef extract and 62.5% peptone. Reference 233000 DIFCO™ BD.
The composition of the yeast extract, as a mass percentage, is 100% autolysate of the yeast Saccharomyces cerevisiae. Reference 210933 DIFCO™ BD.
100 mL of preculture are prepared by suspending a cryotube containing 1 mL of the strain with 100 mL of “inoculum” culture medium at a pH adjusted to 6.8 with 2N NaOH in a 250 mL Fernbach flask and then incubated at 30° C. at 150 rpm for 24 hours. 1.9 L of CM2 “batch” culture medium placed in a presterilized 5 L Fernbach flask are inoculated at OD=0.1 with 100 mL of inoculum.
After 70 hours at 30° C. at 110 rpm, the biomass is dried by lyophilization before being extracted with dichloromethane for 24 hours. The suspension is clarified by filtration on a GF/A filter (Whatman®). The filtrate, composed of PHA dissolved in dichloromethane, is concentrated by evaporation and then dried under high vacuum at 40° C. to constant mass.
The PHA may optionally be purified by successive dissolution and precipitation, for instance using a dichloromethane/methanol system.
The PHA copolymer of Example 17 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure, with: 68 mol % of unit (A) for which R1=isohexenyl and 32 mol % of unit (B) for which R2=isobutyl.
Example 18 is obtained by hydrogenation of the PHA copolymer of Example 17 using an H-Cube Midi® continuous hydrogenator from ThalesNano Technologies.
A solution of 2 g (8.83 mmol) of PHA of Example 17 is prepared with a mixture composed of 100 ml of ethyl acetate (Sigma-Aldrich—CAS: 141-78-6) and 100 mL of methanol (Sigma-Aldrich—CAS: 67-56-1) and is introduced at a flow rate of 3 mL per minute into a hydrogenation cartridge containing the catalyst containing 5% palladium on charcoal (MidiCard ref. DHS 2141; ThalesNano Technologies) maintained at 100° C. under a pressure of 80 bar in the presence of hydrogen in the ThalesNano Technologies H-Cube Midi® system. The reduction of the double bond is monitored by NMR. After six consecutive cycles of reduction, the solution is concentrated by evaporation and then dried under vacuum to constant mass.
The PHA may optionally be purified by successive dissolution and precipitation, for instance using a dichloromethane/methanol system.
The PHA copolymer of Example 18 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure, with: 68 mol % of unit (A) for which R1=isohexyl and 32 mol % of unit (B) for which R2=isobutyl.
A polymer was prepared using the microorganism Pseudomonas putida KT2440 ATCC®47054™ and octanoic acid.
The culture method was performed under batch axenic conditions in 5 L Fernbach flasks (Corning® ref. 431685) containing 2 L of culture medium, shaken at 110 rpm at 30° C. in an orbital incubator (orbit diameter of 2.5 cm). The synthetic process was performed using two different culture media. The first culture medium, defined CM1 “inoculum”, was used for the preparation of the inoculum. The second culture medium, defined CM2 “batch”, was used for unfed batch growth of the microorganism with the octanoic acid in the Fernbach flasks.
The composition in grams per litre of the two media is described in Table 8 below:
100 ml of inoculum were prepared by suspending a cryotube containing 1 ml of the strain with 100 ml of “inoculum” culture medium at a pH adjusted to 6.8 with 2N NaOH in a 250 mL Fernbach flask and then incubated at 30° C. at 150 rpm for 24 hours. 1.9 L of CM2 “batch” culture medium placed in a presterilized 5 L Fernbach flask were inoculated at OD=0.1 with 100 mL of inoculum. After 70 hours at 30° C. at 110 rev/min, the biomass was dried by lyophilization before being extracted with dichloromethane for 24 h. The suspension was clarified by filtration on a GF/A filter (Whatman®). The filtrate, containing the copolymer in solution in the dichloromethane, was concentrated by evaporation and then dried under high vacuum at 40° C. to constant weight. The crude polyhydroxyalkanoate was purified by precipitation of a solution of the latter in solution in 10 times its weight of dichloromethane from 10 volumes of the solution of cold methanol. The solid obtained was dried under high vacuum at 40° C. to constant mass.
The molecular weight of the polyhydroxyalkanoate obtained was characterized by size exclusion chromatography, with detection by refractive index.
The analysis makes it possible to measure the weight-average molecular weight (Mw in g/mol), the number-average molecular weight (Mn in g/mol), the polydispersity index PI (Mw/Mn) and the degree of polymerization DPn.
The monomeric composition of the polyhydroxyalkanoate obtained was defined by gas chromatography equipped with a flame ionization detector. The identification is performed by injection of commercial standards and the monomer composition was determined by a methanolysis and silylation treatment. To determine the monomer composition, 7 mg of the polyhydroxyalkanoate polymer were dissolved in 1.5 mL of chloroform and subjected to methanolysis in the presence of 1.5 mL of an MeOH/HCl solution (17/2, v/v) at 100° C. for 4 hours. The organic phase was then washed with 1 mL of water and then dried over MgSO4. Silylation of the methyl esters formed was performed by adding 100 μL of BSTFA (N,O-bis(trimethylsilyl)trifluoroacetamide) and 100 μL of pyridine to the methylated sample. The solution was heated at 70° C. for 1 hour and then evaporated to dryness. The sample is then dissolved in 600 μL of dichloromethane and analysed by chromatography under the following conditions:
A polymer was prepared using the microorganism Pseudomonas putida KT2440 ATCC®47054™, octanoic acid and acrylic acid.
The culture method was performed under continuous axenic conditions at a dilution D=0.25 h−1 in a 3 L chemostat containing 1.1 L of culture medium. The system was aerated with air at a flow of 3 vvm (vvm=volume of air per volume of fermentation medium per minute) for a nominal dissolved oxygen (OD) value at 30% of saturation.
The production process was performed using three different culture media. The first undefined culture medium (CM1) was used for the preparation of the inoculum. The second defined culture medium (CM2) was used for the unfed batch growth of the microorganism in the fermenter. The third defined culture medium (CM3) was used for the feeding, or maintenance, of the continuous fermentation containing octanoic acid and acrylic acid (inhibitor of the β-oxidation pathway).
The CM1 and CM2 media are identical to those described in example 1. The composition in grams per litre of the medium CM3 is described in Table 10 below:
100 ml of inoculum were prepared by suspending a cryotube containing 1 ml of the strain with 100 ml of Nutrient Broth at a pH adjusted to 7.0 with 2N NaOH in a 250 ml Fernbach flask and were then incubated at 30° C. at 150 rev/min for 24 h.
The fermenter containing 1 litre of culture medium CM2 at 30° C. was inoculated at an optical density of 0.1 at 630 nm (OD 630=0.1). The system was maintained at 30° C. with shaking at 700±200 rpm and regulated in cascade with oxygenation for about 16 hours and/or the time for the microorganism to be able to reach its growth plateau.
Feeding of the fermenter with the medium CM3 was initiated when the microorganism reached its growth plateau, and withdrawal was then performed so as to maintain the initial mass of fermentation medium. Once the equilibrium state was reached in continuous culture, a fraction of the withdrawn material was centrifuged in order to separate the biomass from the fermentation medium. The biomass was dried by lyophilization and then extracted with dichloromethane for 24 hours. The suspension obtained was clarified by filtration through a GF/A filter (Whatman®). The filtrate obtained, comprising the copolymer dissolved in dichloromethane, was concentrated by evaporation and then dried under high vacuum at 40° C. to constant mass. The crude polyhydroxyalkanoate was purified by precipitation of a solution of the latter in solution in 10 times its weight of dichloromethane from 10 volumes of the solution of cold methanol. The solid obtained was dried under high vacuum at 40° C. to constant weight. A copolymer comprising 96% by weight of poly(3-hydroxyoctanoate), 3% by weight of poly(3-hydroxyhexanoate) and 1% by weight of poly(3-hydroxybutanoate) was thus obtained.
A polymer was prepared using the microorganism Pseudomonas putida KT2440 ATCC®47054™, nonanoic acid and acrylic acid. The culture method is performed under continuous axenic conditions at a dilution D=0.25 h−1 in a 3 L chemostat containing 1.1 L of culture medium. The system is aerated with a flow of 1 vvm of air for a nominal dissolved oxygen (OD) value at 30% of saturation. The production process is performed using three different culture media. The first culture medium (CM1) is used for the preparation of the inoculum. The second culture medium (CM2) is used for batch growth of the microorganism in the fermenter. The third culture medium (CM3) is used for the feeding, or maintenance, of the continuous fermentation containing the carbon source of interest and the β-oxidation pathway inhibitor (acrylic acid). The composition in grams per litre of the three media CM1, CM2 and CM3 is described in Table 11 below:
The composition of the Nutrient Broth, as mass percentages, is 37.5% beef extract and 62.5% peptone. Reference 233000 DIFCO™.
The composition of the microelement solution in grams per litre is described in Table 12 below.
100 mL of inoculum are prepared by suspending a cryotube containing 1 mL of the strain at OD=10 with 100 mL of CM1 “inoculum” at a pH preadjusted to 7.0 with 2N NaOH in a 500 mL Fernbach flask and are then incubated at 30° C. at 150 rpm for 24 hours.
The 3 L fermenter containing 1 litre of CM2 “batch” culture medium at 30° C. is inoculated at an optical density of 0.1 at 600 nm (OD 600=0.1). The system is maintained at 30° C. with shaking at 700±200 rpm and regulated in cascade with oxygenation for about 16 hours and/or the time for the microorganism to be able to reach its growth plateau.
Feeding of the fermenter with the CM3 “continuous” medium is initiated when the microorganism has reached its growth plateau, and withdrawal is then performed so as to maintain the initial mass of fermentation medium. Once the equilibrium state is reached in continuous culturing, a fraction of the withdrawn material is centrifuged so as to separate the biomass from the fermentation medium. The biomass is dried by lyophilization and is then extracted with dichloromethane for 24 hours. The suspension is clarified by filtration on a GF/A filter (Whatman®). The filtrate, composed of PHA dissolved in dichloromethane, is concentrated by evaporation and then dried under high vacuum at 40° C. to constant mass.
The molecular weight of the polyhydroxyalkanoate obtained was characterized by size exclusion chromatography, with detection by refractive index.
The analysis makes it possible to measure the weight-average molecular weight (Mw in g/mol), the number-average molecular weight (Mn in g/mol), the polydispersity index PI (Mw/Mn) and the degree of polymerization DPn.
The monomeric composition of the polyhydroxyalkanoate obtained was defined by gas chromatography equipped with a flame ionization detector.
The identification is performed by injection of commercial standards and the monomer composition was determined by a methanolysis and silylation treatment. To determine the monomer composition, 7 mg of the polyhydroxyalkanoate polymer were dissolved in 1.5 mL of chloroform and subjected to methanolysis in the presence of 1.5 mL of an MeOH/HCl solution (17/2, v/v) at 100° C. for 4 hours. The organic phase was then washed with 1 mL of water and then dried over MgSO4. The silylation of the methyl esters formed was carried out by adding 100 μl of BSTFA (N,O-bis(trimethylsilyl)trifluoroacetamide) and 100 μl of pyridine to the methylated sample. The solution was heated at 70° C. for 1 hour and then evaporated to dryness. The sample is then dissolved in 600 μL of dichloromethane and analysed by chromatography under the following conditions:
A polymer was prepared according to the procedure of example 19 using nonanoic acid (instead of octanoic acid) and without acrylic acid.
A copolymer comprising 68% by weight of poly(3-hydroxynonanoate), 27% by weight of poly(3-hydroxyheptanoate) and 5% by weight of poly(3-hydroxypentanoate) was thus obtained.
A polymer was prepared according to the procedure of example 19 using dodecanoic acid (instead of octanoic acid).
A copolymer comprising 44% by weight of poly(3-hydroxydodecanoate), 38% by weight of poly(3-hydroxydecanoate) and 18% by weight of poly(3-hydroxyoctanoate) was thus obtained.
The production process of Example 24 is an adaptation of the article Biomacromolecules 2012, 13, 2926-2932: “Biosynthesis and Properties of Medium-Chain-Length Polyhydroxyalkanoates with Enriched Content of the Dominant Monomer”
The microorganism used is Pseudomonas putida ATCC® 47054™.
The culture method is performed under continuous axenic conditions at a dilution D=0.25 h−1 in a 3 L chemostat containing 1.1 L of culture medium.
The system is aerated with a flow of 3 vvm of air for a nominal dissolved oxygen (OD) value at 30% of saturation.
See
The production process is performed using three different culture media.
The first undefined culture medium (CM1) is used for the preparation of the inoculum.
The second defined culture medium (CM2) is used for batch growth of the microorganism in the fermenter.
The third defined culture medium (CM3) is used for the feeding, or maintenance, of the continuous fermentation containing the carbon source of interest and the β-oxidation pathway inhibitor.
The composition in grams per litre of the three media is described in Table 13. Composition in grams per litre of the culture media for the inoculum and for maintenance.
The composition of the Nutrient Broth, as mass percentages, is 37.5% beef extract and 62.5% peptone. Reference 233000 DIFCO™.
The composition of the microelement solution in grams per litre is described in Table 14: composition in grams per litre of the microelement solution
100 mL of inoculum are prepared by suspending a cryotube containing 1 mL of the strain with 100 mL of Nutrient Broth at a pH adjusted to 7.0 with 2N NaOH in a 250 mL Fernbach flask and are then incubated at 30° C. at 150 rpm for 24 hours.
The fermenter containing 1 litre of culture medium CM2 at 30° C. was inoculated at an optical density of 0.1 at 630 nm (OD 630=0.1). The system is maintained at 30° C. with shaking at 700±200 rpm and regulated in cascade with oxygenation for about 16 hours and/or the time for the microorganism to be able to reach its growth plateau.
Feeding of the fermenter with the medium CM3 is initiated when the microorganism has reached its growth plateau, and withdrawal is then performed so as to maintain the initial mass of fermentation medium. Once the equilibrium state is reached in continuous culturing, a fraction of the withdrawn material is centrifuged so as to separate the biomass from the fermentation medium. The biomass is dried by lyophilization and is then extracted with dichloromethane for 24 hours. The suspension is clarified by filtration on a GF/A filter (Whatman®). The filtrate, composed of PHA dissolved in dichloromethane, is concentrated by evaporation and then dried under high vacuum at 40° C. to constant mass.
The PHA may optionally be purified by successive dissolution and precipitation, for instance using a dichloromethane/methanol system.
The PHA copolymer of Example 24 was fully characterized by spectrometric and spectroscopic methods. By gas chromatography equipped with an FID detector, it is seen that the copolymer contains 96% of radical R1=n-pentyl and 4% of radical R2=n-propyl.
The process for synthesizing the compound of Example 1 is adapted from the article: Fed-batch production of unsaturated medium-chain-length polyhydroxyalkanoates with controlled composition by Pseudomonas putida KT2440, Z. Sun, J.A. Ramsay, M. Guay, B. A. Ramsay, Applied Microbiology Biotechnology, 82. 657-662, 2009.
The microorganism used is Pseudomonas putida KT2440 ATCC® 47054™. The culture method is performed under fed-batch growth axenic conditions with a maintenance solution containing a mixture of carbon source at a rate ρ=0.15 h−1 in a 3 L chemostat containing 2.5 L of culture medium.
The system is aerated with a flow of 0.5 vvm of air for a nominal dissolved oxygen (OD) value at 30% of saturation. The pH is regulated with a solution composed of ammonia and glucose at 15% and 40% final mass, respectively. The temperature of the fermentation medium is regulated at 30° C.
The fermentation medium is regulated in terms of temperature-pressure of dissolved oxygen and pH (not shown).
The production process is performed using three different culture media. The first culture medium, defined CM1 “inoculum”, is used for the preparation of the preculture. The second culture medium, defined CM2 “batch”, is used for unfed batch growth of the microorganism with the primary carbon sources in the Fernbach flasks. The third culture medium, defined CM3 “maintenance”, is used for the fed-batch or maintenance fermentation mode with the carbon sources of interest at a flow rate calibrated as a function of the growth of the microorganism.
The composition of the Nutrient Broth, as mass percentages, is 37.5% beef extract and 62.5% peptone. Reference 233000 DIFCO™.
100 mL of preculture are prepared by suspending a cryotube containing 1 mL of the strain with 100 mL of “inoculum” culture medium at a pH adjusted to 6.8 with 2N NaOH in a 250 mL Fernbach flask and then incubated at 30° C. at 150 rpm for 24 hours. 1.9 L of CM2 “batch” culture medium placed in a presterilized 3 L chemostat are inoculated at OD=0.1 with 100 mL of preculture. After 4 hours at 30° C. at 850 rpm, introduction of the maintenance culture medium is performed, applying the flow rate defined by equation 1.
At the end of the introduction, the biomass is isolated by centrifugation and then washed three times with water. The biomass is dried by lyophilization before being extracted with ethyl acetate for 24 hours. The suspension is clarified by filtration on a GF/A filter (Whatman®). The filtrate, composed of PHA dissolved in the ethyl acetate, is concentrated by evaporation and then dried under high vacuum at 40° C. to constant mass.
The PHA may optionally be purified by dissolution in ethyl acetate and successive precipitations from a 70/30 v/v % ethanol/water system, for example.
The PHA was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure: 95 mol % of unit (B) for which R2=n-hexyl (71%) and n-butyl (24%) and 5 mol % of unit (A) for which R1=8-bromo-n-octanyl (5.9%) and 6-bromo-n-hexyl (0.2%).
The compounds of Examples 1 to 25 may be mixed with one or more oligo/polyesters b) as defined previously; preferably in the presence of a liquid fatty substance c) such as isododecane and optionally water e). The mixing of the PHA(s) a) with the oligo/polyester(s) b) may be performed at room temperature, with stirring, preferably in the presence of a liquid fatty substance c) and optionally of organic solvent(s) d) other than c) as defined previously. According to one variant, water e) is added to the mixture of a), b) and c) and one or more organic solvents d) other than c) as defined previously are then optionally added.
Compositions 26 (comparative) and 27 to 32 (invention) described in Tables 17 and 18 below were prepared:
The PHA, the fatty substance and ethanol are stirred at 2500 rpm, at a temperature of 25° C. The oligo/polyester is introduced and the medium is heated from 25° C. to 80° C. with stirring at 2500 rpm. The medium is maintained at 80° C. for 30 minutes with stirring at 3000 rpm and is then cooled from 80° C. to 25° C. with stirring at 2500 rpm.
The first step in this test consists in making a deposit. The deposits are prepared on a Byko Chart Lenata contrast card using a film spreader and left to dry for 24 hours at 25° C. and 45% RH. The final thickness of the deposit is 30 μm.
A wear resistance test is performed on this dry deposit. A hydrophilic steel ball is used as a friction device. The load or normal force applied is 1N, and the displacement speed is 50 mm·s−1. On each film are defined tracks on which the friction device makes multiple passes. In the case of wear measurements, permanent contact is maintained during the to and fro trips of the ball on the deposit. The number of passes is increased for each track. The wear resistance is quantified as the minimum number of passes to completely wear out the deposit.
In the case of this study, the number of passes per track are, respectively, 10, 30, 50, 100, 200 and 300 passes.
Each measurement was repeated five times.
The results of the wear resistance tests are quantified as described in the table below:
It is seen from the resistance tests that the substrate which was treated with the compositions of the invention makes it possible to significantly improve the wear resistance relative to the comparative composition free of oligo/polyester b).
On the same 30 μm dry deposit identical to that made for the wear test, the sensitivity to stressors is evaluated after depositing a drop of stressor (20 μl for water) on the surface of the deposit. The evaluations are made after 1 hour of contact between the stressor and the deposit. The level of sensitivity to stressors is noted as follows. It is seen that the compositions of the invention (Ex. 27 to 33) are highly resistant to water since the film remained intact.
Compositions 33 and 34 (Invention) Described in Table 21 Below were Prepared:
The first step in this test consists in making a deposit. The deposits are prepared on a Byko Chart Lenata contrast card using a film spreader and left to dry for 24 hours at 25° C. and 45% RH. The final thickness of the deposit is 30 μm.
The films obtained from the compositions of Examples 33 and 34 are satisfactory in terms of wear and water-resistance.
It has been observed that the deposit obtained from the composition containing 20% oligoester (Example 34) is significantly stickier than the deposit obtained from the composition according to the invention containing 15% oligoester (Example 33).
| Number | Date | Country | Kind |
|---|---|---|---|
| FR2114369 | Dec 2021 | FR | national |
This is a National Stage Application of PCT/EP2022/087639, filed internationally on Dec. 22, 2022, which claims priority to French Application No. 2114369, filed on Dec. 23, 2021, which are incorporated by reference herein in their entireties.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2022/087639 | 12/22/2022 | WO |
