Patent Application
20240415160
The present disclosure relates to the field of food processing, in particular to a composition, and preparation method therefor and application thereof.
Kluyveromyces is a kind of ascosporogenous yeast, which is a food-safe grade yeast. Among them, Kluyveromyces marxianus and Kluyveromyces lactis are the yeast which are widely used in industry and studied. For example, Kluyveromyces marxianus is widely found in yogurt, fruits and kefir. Due to the characteristics of high food safety, high growth rate, high biomass and high temperature resistance this yeast showed, it has been widely used in fermentation, development of lactobacillus beverages and other fields. However, the nutritional, physical and chemical properties of inactivated Kluyveromyces itself are less studied.
Emulsification technique is the most common process in the food field, especially in the production process of beverages and sauces. In order to obtain uniform and stable products without stratification, it is usually necessary to use emulsification technique to mix the water phase and oil phase evenly, and add a certain amount of one or more emulsifier composition, so that the different phases cannot repel too much and the same phase cannot gather easily between the two incompatible phases to ensure the long-term stability of the system, thereby ensuring the shelf life of the products. Emulsifier can be classified as a surfactant in the chemistry field, and a food additive in the food field. There are three types of emulsifiers commonly used in the processing of beverages and sauces: ionic, non-ionic and ampholytic. The characteristics and functions of these three types of emulsifiers are usually determined by the relative strength of the hydropathy of the hydrophilic groups and the hydrophobicity of the lipophilic groups in their molecules. A good emulsifier system must be fairly balanced between the hydrophilic groups and the hydrophobic groups. In the present food processing technology, emulsifiers are used to obtain a stable and uniform emulsification system, which usually requires strict theoretical calculation to balance the hydrophilic and hydrophobic groups in the emulsification system. Due to the diversification of food formulas, beyond theory it usually needs to rely on experience, or many experiments, or even simple and crude excessive use of emulsifiers to obtain a relative stable formula system. Therefore, it is an urgent industry problem need to develop a natural and simple technique which can make food containing both water phase and oil phase simultaneously maintain long-term stable and uniform.
The problem solved in the present disclosure is to provide a food composition without food additives, which is stable and uniform, has high nutritional components such as protein and dietary fiber.
The present disclosure also provides applications of the food composition in the preparation of food products.
In order to achieve the above purposes, the present disclosure provides the following solutions: In one aspect, the present disclosure provides a composition comprising the following raw materials in weight percentage: 2.5%-80% of preceding inactivated Kluyveromyces cell, 1%-50% of edible oil, and 18%-96.5% of water. According to the studies in the present disclosures, the composition and proportion of the raw material is conducive to the formation of a stable and uniform emulsification system, wherein inactivated Kluyveromyces cells can physically isolate the mutual contact between the oil particles. In the case of no exogenous food additives such as emulsifiers, stabilizers and thickeners, they can be directly applied to the emulsification technique of water phase and oil phase to obtain a stable and uniform emulsification system, which can maintain for at least 7 days without demulsification and stratification.
In an embodiment in the present disclosure, the weight percentage of the preceding inactivated Kluyveromyces cell is 10-70%.
In an embodiment in the present disclosure, the weight percentage of the preceding edible oil is 10-40%.
In an embodiment in the present disclosure, the preceding composition may comprise the following raw materials in weight percentage: 2.5-25% of inactivated Kluyveromyces cell, 1-40% of edible oil, and 50-96.5% of water. For example, it can comprise 2.5%, 5%, 10%, 15%, 20% and 25% of inactivated Kluyveromyces cell, 1%, 2%, 4%, 8%, 12%, 16%, 20%, 25%, 30%, 35% and 40% of edible oil. The composition and proportion of the raw material is conducive to it maintaining for at least 28 days without demulsification and stratification.
In the implementation of the present disclosure, when the content of inactivated Kluyveromyces cells is 2.5-25%:
The frequency of components with particle size≤55 μm is 5-98%. For example, it can be 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%.
The frequency of components with particle size≤10 μm is 8-100%. For example, it can be 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%.
The frequency of components with particle size≤50 μm is 38-100%. For example, it can be 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%.
The frequency of components with particle size≤100 μm is about 100%.
This frequency distribution helps maintain the long-term stability of the composition system.
In a preferred embodiment in the present disclosure, the preceding composition comprises the following raw materials in weight percentage: 30-70% of inactivated Kluyveromyces cell, 1-30% of edible oil, and 18-69% of water. For example, it can comprise 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% and 70% of inactivated Kluyveromyces cell, 1%, 2%, 4%, 8%, 12%, 16%, 20%, 25% and 30% of edible oil. The composition and proportion of the raw material is conducive to it maintaining for at least 28 days without demulsification and stratification.
In the implementation of the present disclosure, when the content of inactivated Kluyveromyces cells is 30-70%:
The frequency of components with particle size≤5 μm is 54-100%. For example, it can be 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%.
The frequency of components with particle size≤10 μm is 55-100%. For example, it can be 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%.
The frequency of components with particle size≤50 μm is 75-100%. For example, it can be 75%, 80%, 85%, 90%, 95%.
The frequency of components with particle size≤100 μm is about 100%.
This frequency distribution helps maintain the composition system without demulsification and stratification for at least 28 days.
According to the studies in the present disclosure, the particle size of the inactivated Kluyveromyces cell in the preceding composition can be 1-7 μm, such as 1 μm, 2 μm, 3 μm, 4 μm, 5 μm, 6 μm and 7 μm, which is conducive to isolate the mutual contact between the oil particles in the form of “caulking” in the oil-in-water (the proportion of water is greater than the proportion of oil) system, or enrich in water by polarity to form microparticles to isolate the mutual contact between the oil particles in the water-in-oil (the proportion of oil is greater than the proportion of water) system, thereby delaying the demulsification time of the composition system.
In a preferred embodiment in the present disclosure, the preceding composition is an oil-in-water system.
Specifically, the preceding inactivated Kluyveromyces cell is selected from one or more of Kluyveromyces marxianus, Kluyveromyces lactis, Kluyveromyces hubeiensis, Kluyveromyces wickerhamii, and Kluyveromyces thermotolerans cell. According to the studies in the present disclosure, the selection of these 5 yeast cells is conducive to obtaining the inactivated Kluyveromyces cells with the preceding particle size range.
Preferably, the preceding inactivated Kluyveromyces cell is Kluyveromyces marxianus and/or Kluyveromyces lactis cell. Kluyveromyces marxianus is an edible strain published by the National Health Commission. Kluyveromyces lactis is a strain published by the National Health Commission that can be used for health food.
In an preferable embodiment in the present disclosure, the particle size of the inactivated Kluyveromyces cell in the preceding composition can be 2-5 μm, which is more conducive to isolating the mutual contact between the oil particles in the composition system where the components of water phase and oil phase coexist, thereby delaying demulsification time.
According to the further studies in the present disclosure, when the preceding inactivated Kluyveromyces cell is Kluyveromyces hubeiensis cell, the particle size thereof can be 2-5 μm. For example, it can be 2 μm, 3 μm, 4 μm, 5 μm.
When the preceding inactivated Kluyveromyces cell is Kluyveromyces wickerhamii cell, the particle size thereof can be 2-7 μm. For example, it can be 2 μm, 3 μm, 4 μm, 5 μm, 6 μm, 7 μm.
When the preceding inactivated Kluyveromyces cell is Kluyveromyces thermotolerans cell, the particle size thereof can be 2-5 μm. For example, it can be 2 μm, 3 μm, 4 μm, 5 μm.
When the preceding inactivated Kluyveromyces cell is Kluyveromyces marxianus cell, the particle size thereof can be 2-7 μm. For example, it can be 2 μm, 3 μm, 4 μm, 5 μm, 6 μm, 7 μm.
When the preceding inactivated Kluyveromyces cell is Kluyveromyces lactis cell, the particle size thereof can be 2-7 μm. For example, it can be 2 μm, 3 μm, 4 μm, 5 μm, 6 μm, 7 μm.
According to the studies in the present disclosure, the particle size of the edible oil in the preceding composition can be 1-100 μm. Controlling the particle size of the edible oil in the composition within the range of 1-100 μm is more conducive to achieving the balance of processing technique and product stability. If the particle size of the edible oil in the composition is too large, it is difficult for inactivated Kluyveromyces cells to play an effective physical isolation role, so that the oil particles combine into large oil droplets quickly, which leads to demulsification and stratification of the system and effects on stability of the products.
In a specific embodiment in the present disclosure, the particle size of the edible oil in the preceding composition is 5-100 μm, and can further be 5-10 μm, 5-50 μm, 10-50 μm, 10-100 μm and 50-100 μm. For example, it can be 5 μm, 10 μm, 15 μm, 20 μm, 25 μm, 30 μm, 35 μm, 40 μm, 45 μm, 50 μm, 55 μm, 60 μm, 65 μm, 70 μm, 75 μm, 80 μm, 85 μm, 90 μm, 95 μm.
The particle size of the edible oil in the composition is calculated by the average particle size of compositions in the system, the average particle size of yeast cells and the particle size frequency of edible oil. The average particle size of the yeast cells added in the composition, the proportion of each component, and the total frequency of particle size distribution in the system are known.
The particle size frequency of edible oil=the total frequency of particle size distribution in the system−the average particle size frequency of the yeast cells.
Specifically, the preceding inactivated Kluyveromyces cell comprises 25-55% of protein, 1-5% of fat, and 15-30% of dietary fiber. It is characterized by high protein, low fat (including saturated fat and trans fat), and rich soluble dietary fiber, which can be applied in food as a nutritional source.
In a preferred embodiment in the present disclosure, the preceding inactivated Kluyveromyces cell comprises 30.5-52.5% of protein, 2.1-4.8% of fat, and 15.5-29.0% of dietary fiber.
Specifically, the preceding inactivated Kluyveromyces cell comprises 6-10% of water, and 2.5-10.5% of ash.
In a embodiment in the present disclosure, the preceding inactivated Kluyveromyces cell can be dry inactivated Kluyveromyces cell mycelium (inactivated Kluyveromyces cell dry powder), dehydrated inactivated Kluyveromyces cell, or inactivated Kluyveromyces cell mycelium suspension.
In a preferred embodiment in the present disclosure, the preceding inactivated Kluyveromyces cell is inactivated Kluyveromyces cell mycelium suspension.
In a preferred embodiment in the present disclosure, the preceding inactivated Kluyveromyces cell is dry inactivated Kluyveromyces cell mycelium. When it is dry Kluyveromyces cell mycelium, it is more convenient for the preservation, transportation, storage and use of the preceding composition.
In an embodiment in the present disclosure, the edible oil in the present disclosure is selected from one or more of soybean oil, rapeseed oil, high oleic sunflower seed oil, medium chain triglyceride, canola oil, coconut oil, corn oil, sesame oil, tea seed oil, rice bran oil, olive oil, linseed oil, safflower seed oil, grape seed oil, walnut oil, palm oil, peanut oil, and blend oil.
In a preferred embodiment in the present disclosure, the edible oil in the present disclosure is selected from one or more of coconut oil, medium chain triglyceride, canola oil, high oleic sunflower seed oil, olive oil, and rapeseed oil.
The preceding inactivated Kluyveromyces cell can be prepared by the method comprising the following steps:
Specifically, the preceding medium containing carbon source, nitrogen source and salts may comprise but not be limited to one or more carbon source, nitrogen source and salts composition selected from molasses, glucose, starch, dipotassium phosphate, corn pulp dry powder, sodium hydroxide, magnesium sulfate, ammonium sulfate, ammonia, urea, sodium chloride, yeast extract, peptone, potassium dihydrogen phosphate, potassium hydroxide, methionine, cysteine, alanine, glycine and glutamic acid.
In an embodiment in the present disclosure, the preceding medium containing carbon source, nitrogen source and salts can comprise glucose, magnesium sulfate, ammonium sulfate, yeast extract, potassium dihydrogen phosphate. Specifically, the weight percentage of each component thereof can be 3.5-4.5% of glucose, 0.2-7% of molasses, 0.1-0.3% of corn pup dry powder, 0.02-0.15% of magnesium sulfate, 0.5-0.6% of ammonium sulfate, 0.6-0.9% of yeast extract, 0.4-0.6% of potassium dihydrogen phosphate. It can also comprise other inorganic salt components such as copper sulfate, ferrous sulfate, manganese sulfate, cobalt chloride, zinc sulfate, and other nutrients such as methionine, alanine, cysteine, glycine, the rest is water.
Specifically, the culture time of the preceding inactivated Kluyveromyces in the medium can be 15-40 h. For example, it can be 15 h, 20 h, 25 h, 30 h, 35 h or 40 h.
Specifically, the pH of the preceding medium can be 4.0-8.0. For example, it can be 4, 4.5, 5.0, 5.5, 6, 6.5, 7, 7.5, 8.
Specifically, the preceding fermentation temperature can be 25-50° C. For example, it can be 25-30° C., 25-35° C., 25-40° C., 25-45° C., 30-35° C., 30-40° C., 30-45° C., 30-50° C., 35-40° C., 35-45° C., 35-50° C., 40-45° C., 45-50° C.
Specifically, the preceding fermentation time can be 15-40 h. For example, it can be 15 h, 20 h, 22 h, 24 h, 26 h, 28 h, 30 h, 32 h, 36 h, 40 h.
In an embodiment in the present disclosure, the preceding dry inactivated Kluyveromyces cell mycelium, dehydrated inactivated Kluyveromyces cell, and inactivated Kluyveromyces cell mycelium suspension are prepared by the following methods:
Selecting a medium containing carbon source, nitrogen source and salts to culture the Kluyveromyces strain for 15-40 hours; regulating medium pH to 4.0-8.0, heating and inactivating after fermentation at 25-50° C. for 15-40 h, and obtaining the inactivated Kluyveromyces pulp after removing the supernatant by centrifugation; post-treating the the Kluyveromyces pulp to obtain treated pulp, which is inactivated Kluyveromyces cell mycelium suspension.
Then drying the treated pulp to obtain dry inactivated Kluyveromyces cell mycelium, and dehydrating to obtain dehydrated inactivated Kluyveromyces cell.
In the implementation of the present disclosure, the drying treatment is commonly used drying techniques in the field such as freeze drying, spray drying and baking drying. The dehydrating treatment is commonly used technique in the field. It is not specifically limited in the present disclosure.
Specifically, the preceding heating and inactivation can be implemented at 100° C.; the time of heating and inactivation can be longer than 20 min. For example, it can be 20 min, 25 min, 30 min, 35 min, 40 min.
The preceding post-treatment process includes one or more selected from water cleaning, pH regulation, alcohol precipitation, alcohol extraction, activated carbon adsorption, ozone treatment, protease treatment, cellulase treatment, hemicellulase treatment, lipase treatment, freezing treatment, solution pressurization treatment and heating treatment. The post-treatment process is commonly used technique in the field. It is not specifically limited in the present disclosure.
The preceding protease can be endopeptidase or exopeptidase, which can be derived from microorganisms, plants or animals. For example, it can be serine protease, cysteine protease, aspartic protease, protease derived from microorganisms, papain and bromelain derived from plants, trypsin, pepsin and cathepsin derived from animals, etc. The preceding cellulase is an enzyme that can degrade cellulose to produce glucose, such as β-1,4-glucan-4-glucan hydrolase.
In another aspect, the present disclosure provides a preparation method of the preceding composition, comprising:
The proportion of inactivated Kluyveromyces cell in the composition described in the present disclosure is not affected by the form of inactivated Kluyveromyces cell mycelium added. Understandably, when the inactivated Kluyveromyces cell used is inactivated Kluyveromyces cell dry powder, it can be added directly according to the preceding proportion. When the inactivated Kluyveromyces cell used is inactivated Kluyveromyces cell mycelium suspension, the preceding proportion of inactivated Kluyveromyces cell in the composition can be calculated by number of cells.
In another aspect, the present disclosure provides a food product comprising the preceding composition.
In an embodiment in the present disclosure, when the content of inactivated Kluyveromyces cell is 2.5-25%, the composition is beverage, wherein comprises 2.5-25% of inactivated Kluyveromyces cell, 1-40% of edible oil, and 40-96.5% of water.
In an embodiment in the present disclosure, when the content of inactivated Kluyveromyces cell is 30-70%, the composition is sauce, wherein comprises 30-70% of inactivated Kluyveromyces cell, 1-30% of edible oil, and 18-69% of water.
In another aspect, the present disclosure provides an application of the preceding composition in the preparation of food products.
In an embodiment in the present disclosure, the preceding composition is preferably used in the preparation of biscuits, bread, baked goods, puffed food, freeze-dry food, ice cream, and dehydrated food. The composition in the present disclosure has a high protein content, which can be added to food products as a protein substitute. Meanwhile, the food composition in the present disclosure characterized by low fat (saturated fat, trans fat), and rich soluble dietary fiber, which has good food value.
In the implementation of the present disclosure, when the preceding beverage of food composition is used to prepare food products, the amount added thereof is greater than 10%. When the preceding sauce of food composition is used to prepare food products, the amount added thereof is greater than 3%.
In order to illustrate the technology solutions in the embodiments in the present disclosure or in the prior art more clearly, the drawings required to be used in the description of the embodiments or the prior art are briefly described below. Obviously, the drawings described below are only one embodiment of the present disclosure. Other embodiments can be obtained according to these drawings for those skilled in the art.
In the present disclosure, unless otherwise specified, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Moreover, protein and nucleic acid chemistry, molecular biology, cell and tissue culture, microbiology, and immunology-related terms and laboratory procedures used herein are terms and conventional procedures widely used in corresponding arts. Meanwhile, in order to better understand the present disclosure, definitions and explanations of relevant terms are provided below.
For the purpose of a clear and concise description, characteristics are described herein as a part of identical or separate embodiments. However, understandably, the scope of the present disclosure may include embodiments having a combination of all or some of the characteristics described.
As used herein, unless otherwise specified, the term “about” or “approximate” means within plus or minus 10% of a given value or range. In the case that integers are required, the term means rounding up or down to the nearest integer within plus or minus 10% of a given value or range.
As used herein, unless otherwise specified, the term “comprise” “include” “have” “contain”, including their grammatical equivalents, should generally be understood to be open and non-limited, such as not excluding other elements or steps not mentioned.
As used herein, the term “fermentation” refers to the process by which specific metabolic pathways in biological cells are used to transform external substrates to produce target products or mycelia needed by people.
The term “shear emulsifying” refers to that the materials are thrown radially to the narrow precise gap between the stator and the rotor under the action of centrifugal force produced by high-speed and strongly rotating rotor, which meanwhile under the action of centrifugal extrusion, impact and other forces, and/or high-pressure homogenizing treatment added in the later stage so that materials can be evenly dispersed, mixed and emulsified.
The term “food additive” means any artificially synthetic or natural substance put into food to improve its quality, color, fragrance or taste, or for the sake of preservation, fresh-keeping and processing. Spices for food, bodying agent in gum base candies, and processing AIDS for food industry are also included.
The term “drying” refers to the operation of using heat energy to vaporize the wet component of a wet material, and using air flow or vacuum to carry away the vaporized wet component, thereby obtaining a dry material.
Kluyveromyces CJ3113 used in the embodiments in the present disclosure is preserved in the China Center for Type Culture Collection (CCTCC). The preservation address is the Collection Center of Wuhan University, Wuhan City, Hubei Province. The preservation number is CCTCC No: M20211265, whose Latin scientific name is Kluyveromyces marxianus. The preservation date is Oct. 13, 2021.
Kluyveromyces lactis was bought from China Center of Industrial Culture Collection (CICC). The preservation number is CICC 32428.
Kluyveromyces hubeiensis, Kluyveromyces wickerhamii and Kluyveromyces thermotolerans was bought from China General Microbiological Culture Collection Center (CGMCC). The preservation numbers are CGMCC 2.4330, CGMCC 2.4309 and CGMCC 2.4072, respectively.
By observation and measurement through scanning electron microscope (ZEISS GeminiSEM500, field emission), the minimum (μm) and maximum (μm) particle size with stable effect of inactivated Kluyveromyces marxianus, inactivated Kluyveromyces lactis, inactivated Kluyveromyces hubeiensis, inactivated Kluyveromyces wickerhamii and inactivated Kluyveromyces thermotolerans were obtained, which are shown in Table 1. The measurement schematics by scanning electron microscope are shown in
Kluyveromyces
hubeiensis
Kluyveromyces
wickerhamii
Kluyveromyces
thermotolerans
Kluyveromyces
marxianus
Kluyveromyces lactis
Kluyveromyces hubeiensis strain was cultured in the medium containing carbon source, nitrogen source and salts (4.5% of glucose, 0.2% of molasses, 0.2% of corn pulp dry powder, 0.02% of magnesium sulfate, 0.6% of ammonium sulfate, 0.6% of yeast extract, 0.6% of potassium dihydrogen phosphate, 6 ppm copper sulfate, 12 ppm ferrous sulfate, 15 ppm manganese sulfate, 6 ppm cobalt chloride, 20 ppm zinc sulfate, 8 ppm methionine, 5 ppm alanine, 2 ppm cysteine, 5 ppm glycine); regulating medium pH to 4.0-5.0, and heating at 100° C. for 20 min after fermentation at 25-30° C. for 40 h, the Kluyveromyces pulp was obtained after removing the supernatant by centrifugation; the Kluyveromyces pulp was post-treated as follows: the pulp was obtained after centrifugation, soaked in deionized water five times the pulp for 30 min, and adjusted pH to 5.0 by food-grade lactic acid. 0.01% of acid protease and 0.01% of papain by weight of the pulp was added to react at 50° C. for 12 h. Particle size was measured by laser particle size analyzer. The pH was adjusted to 6.6-6.8 by food-grade sodium bicarbonate. 0.02% of β-glucolase and 0.01% of alkaline protease by weight of the pulp was added to react at 55° C. for 36 h. Particle size was measured by laser particle size analyzer. Adsorbing and decolorizing by activated carbon, the inactivated Kluyveromyces cell mycelium suspension was obtained. The suspension was frozen at −20° C. for 1 h cooling treatment, and soaked to sterilize for 60 min in 85% ethanol solution which was three times the weight of the pulp. The deionized water one times the weight of the pulp was mixed after recovering ethanol. The solution was sterilized for 1 h by passing ozone in. The Kluyveromyces hubeiensis cell dry powder was obtained by spray drying after pasteurization or pressure sterilization.
Kluyveromyces wickerhamii strain was cultured in the medium containing carbon source, nitrogen source and salts (4.2% of glucose, 0.5% of molasses, 0.1% of corn pulp dry powder, 0.03% of magnesium sulfate, 0.5% of ammonium sulfate, 0.6% of yeast extract, 0.4% of potassium dihydrogen phosphate, 5 ppm copper sulfate, 10 ppm ferrous sulfate, 10 ppm manganese sulfate, 5 ppm cobalt chloride, 18 ppm zinc sulfate, 5 ppm methionine, 6 ppm alanine, 3 ppm cysteine, 1 ppm glycine); regulating medium pH to 4.0-5.0, and heating at 100° C. for 30 min after fermentation at 45-50° C. for 15 h, the Kluyveromyces pulp was obtained after removing the supernatant by centrifugation; the Kluyveromyces pulp was post-treated as follows: the pulp was obtained after centrifugation, soaked in deionized water five times the pulp for 60 min, and adjusted pH to 5.0 by food-grade lactic acid. 0.02% of acid protease and 0.015% of papain by weight of the pulp was added to react at 50° C. for 6 h. Particle size was measured by laser particle size analyzer. The pH was adjusted to 6.6-6.8 by food-grade sodium bicarbonate. 0.015% of β-glucolase and 0.015% of alkaline protease by weight of the pulp was added to react at 55° C. for 24 h. Particle size was measured by laser particle size analyzer. Adsorbing and decolorizing by activated carbon, the inactivated Kluyveromyces cell mycelium suspension was obtained. The suspension was frozen at −20° C. for 1 h cooling treatment, and soaked to sterilize for 60 min in 85% ethanol solution which was three times the weight of the pulp. The deionized water one times the weight of the pulp was mixed after recovering ethanol. The solution was sterilized for 1 h by passing ozone in. The Kluyveromyces wickerhamii cell dry powder was obtained by spray drying after pasteurization or pressure sterilization.
Kluyveromyces thermotolerans strain was cultured in the medium containing carbon source, nitrogen source and salts (3.5% of glucose, 0.7% of molasses, 0.3% of corn pulp liquor dry powder, 0.02% of magnesium sulfate, 0.6% of ammonium sulfate, 0.7% of yeast extract, 0.4% of potassium dihydrogen phosphate, 1 ppm copper sulfate, 3 ppm ferrous sulfate, 15 ppm manganese sulfate, 3 ppm cobalt chloride, 10 ppm zinc sulfate, 15 ppm methionine, 3 ppm alanine, 1 ppm cysteine, 9 ppm glycine); regulating medium pH to 7.5-8.0, and heating at 100° C. for 25 min after fermentation at 30-35° C. for 30 h, the Kluyveromyces pulp was obtained after removing the supernatant by centrifugation; the Kluyveromyces pulp was post-treated as follows: the pulp was obtained after centrifugation, soaked in deionized water five times the pulp for 30 min, and adjusted pH to 5.0 by food-grade lactic acid. 0.01% of acid protease and 0.01% of papain by weight of the pulp was added to react at 50° C. for 12 h. Particle size was measured by laser particle size analyzer. The pH was adjusted to 6.6-6.8 by food-grade sodium bicarbonate. 0.02% of β-glucolase and 0.01% of alkaline protease by weight of the pulp was added to react at 55° C. for 30 h. Particle size was measured by laser particle size analyzer. Adsorbing and decolorizing by activated carbon, the inactivated Kluyveromyces cell mycelium suspension was obtained. The suspension was frozen at −20° C. for 1 h cooling treatment, and soaked to sterilize for 60 min in 85% ethanol solution which was three times the weight of the pulp. The deionized water one times the weight of the pulp was mixed after recovering ethanol. The solution was sterilized for 1 h by passing ozone in. The Kluyveromyces thermotolerans cell dry powder was obtained by spray drying after pasteurization or pressure sterilization.
Kluyveromyces marxianus strain was cultured in the medium containing carbon source, nitrogen source and salts (4.5% of glucose, 0.2% of molasses, 0.2% of corn pulp dry powder, 0.05% of magnesium sulfate, 0.5% of ammonium sulfate, 0.8% of yeast extract, 0.6% of potassium dihydrogen phosphate, 10 ppm copper sulfate, 5 ppm ferrous sulfate, 15 ppm manganese sulfate, 7 ppm cobalt chloride, 2 ppm zinc sulfate, 3 ppm methionine, 1 ppm alanine, 6 ppm cysteine, 3 ppm glycine); regulating medium pH to 5.5-6.5, and heating at 100° C. for 30 min after fermentation at 25-30° C. for 24 h, the Kluyveromyces pulp was obtained after removing the supernatant by centrifugation; the Kluyveromyces pulp was post-treated as follows: the pulp was obtained after centrifugation, soaked in deionized water five times the pulp for 30 min, and adjusted pH to 5.0 by food-grade lactic acid. 0.015% of acid protease and 0.015% of papain by weight of the pulp was added to react at 50° C. for 6 h. Particle size was measured by laser particle size analyzer. The pH was adjusted to 6.6-6.8 by food-grade sodium bicarbonate. 0.01% of β-glucolase and 0.02% of alkaline protease by weight of the pulp was added to react at 55° C. for 24 h. Particle size was measured by laser particle size analyzer. Adsorbing and decolorizing by activated carbon, the inactivated Kluyveromyces cell mycelium suspension was obtained. The suspension was frozen at −20° C. for 1 h cooling treatment, and soaked to sterilize for 60 min in 85% ethanol solution which was three times the weight of the pulp. The deionized water one times the weight of the pulp was mixed after recovering ethanol. The solution was sterilized for 1 h by passing ozone in. The Kluyveromyces marxianus cell dry powder was obtained by spray drying after pasteurization or pressure sterilization.
Kluyveromyces lactis strain was cultured in the medium containing carbon source, nitrogen source and salts (4.2% of glucose, 0.5% of molasses, 0.1% of corn pulp dry powder, 0.15% of magnesium sulfate, 0.6% of ammonium sulfate, 0.9% of yeast extract, 0.5% of potassium dihydrogen phosphate, 5 ppm copper sulfate, 10 ppm ferrous sulfate, 10 ppm manganese sulfate, 5 ppm cobalt chloride, 18 ppm zinc sulfate, 5 ppm methionine, 6 ppm alanine, 3 ppm cysteine, 1 ppm glycine); regulating medium pH to 4.0-5.0, and heating at 100° C. for 20 min after fermentation at 40-45° C. for 28 h, the Kluyveromyces pulp was obtained after removing the supernatant by centrifugation; the Kluyveromyces pulp was post-treated as follows: the pulp was obtained after centrifugation, soaked in deionized water five times the pulp for 60 min, and adjusted pH to 5.0 by food-grade lactic acid. 0.02% of acid protease and 0.02% of papain by weight of the pulp was added to react at 50° C. for 6 h. Particle size was measured by laser particle size analyzer. The pH was adjusted to 6.6-6.8 by food-grade sodium bicarbonate. 0.01% of β-glucolase and 0.015% of alkaline protease by weight of the pulp was added to react at 55° C. for 36 h. Particle size was measured by laser particle size analyzer. Adsorbing and decolorizing by activated carbon, the inactivated Kluyveromyces cell mycelium suspension was obtained. The suspension was frozen at −20° C. for 1 h cooling treatment, and soaked to sterilize for 60 min in 85% ethanol solution which was three times the weight of the pulp. The deionized water one times the weight of the pulp was mixed after recovering ethanol. The solution was sterilized for 1 h by passing ozone in. The Kluyveromyces lactis cell dry powder was obtained by spray drying after pasteurization or pressure sterilization.
The compositions of protein, fat, dietary fiber, water, ash and total nitrogen in the Kluyveromyces cell dry powder prepared in Example 2-6 were analyzed. Water content was determined by atmospheric dry weight method (105° C., 3 h). Total nitrogen was determined by Kjeldahl method. Ash content was determined by Muffle furnace direct ashing method. Dietary fiber content was determined by AOAC 991.43 standard. Protein content was determined by AOAC 979.09 standard. Fat content was determined by AOAC 996.06 standard. Water content was determined by AOAC 925.09 method. Ash content was determined by AOAC 942.05 method. The analysis results are shown in Table 2:
The inactivated Kluyveromyces cell dry powder prepared in Example 5 was used as a food raw material for the development of food formula, which could partially or completely substitute (the use of) emulsifier, thickener and stabilizer in food.
The food composition comprising inactivated Kluyveromyces cells has characters of stable and uniform (no stratification, no demulsification) without the use of exogenous emulsifier, thickener, and stabilizer. Specifically, it can be divided into three characters (A is defined as no demulsification and no stratification at all; B is defined as slight demulsification and a little amount of stratification at the top of the product; C is defined as obvious demulsification and a complete oil and water stratification of product) to analyze:
Finding the method with an appropriate proportion of water and oil based on the fixed yeast addition, it is supposed that the theoretical stable status can be reached when the contents of inactivated Kluyveromyces cell, oil and water in the composition in the present disclosure are as follows:
Kluyveromyces cell
Beverage of food with required particle size distribution was prepared by emulsification in a certain weight percentage of inactivated Kluyveromyces cell dry powder prepared in Example 2-6, rapeseed oil and water. The specific steps are as follows:
The results of the percentage content by weight of inactivated Kluyveromyces cell dry powder, rapeseed oil and water, the particle size distribution of composition, and the results of stability test are shown in Table 4, Table 5, Table 6, Table 7, and Table 8 (A is defined as no demulsification and no stratification at all; B is defined as slight demulsification and a little amount of stratification at the top of the product; C is defined as obvious demulsification and a complete oil and water stratification of product):
As shown in Table 4-Table 8, after shear emulsifying process until the particle sizes of each component in the suspension are all less than or equal to 100 μm (the frequency of particle size≤100 mμm is 100%), all can maintain a stable and uniform status without stratification for 7 days at room temperature when the proportion of inactivated Kluyveromyces cell powder is 2.5-25%, the proportion of edible oil is 1%-40%, and the proportion of water is 40%-96.5%; it can even maintain a stable and uniform status without stratification for 28 days when the proportion of inactivated Kluyveromyces cell powder is 2.5-25%, the proportion of edible oil is 1%-30%, the proportion of water is 40%-96.5% and the particle size of each component in the suspension is less than or equal to 100 μm by the same shearing.
Sauce of food with required particle size distribution was prepared by emulsification in a certain weight percentage of inactivated Kluyveromyces cell dry powder prepared in Example 2-6, rapeseed oil, and water. The specific steps are as follows:
The results of the percentage content by weight of inactivated Kluyveromyces cell dry powder, rapeseed oil and water, the particle size distribution of composition, and the results of stability test are shown in Table 9, Table 10, Table 11, Table 12, and Table 13 (A is defined as no demulsification and no stratification at all; B is defined as slight demulsification and a little amount of stratification at the top of the product; C is defined as obvious demulsification and a complete oil and water stratification of product):
As shown in Table 9-Table 13, after shear emulsifying process until the particle sizes of each component in the suspension are all less than or equal to 100 μm (the frequency of particle size≤100 mμm is 100%), all can maintain a stable and uniform status without stratification for 28 days at room temperature when the proportion of inactivated Kluyveromyces cell powder is 30-70%, the proportion of edible oil is 1%-30%, and the proportion of water is 18%-69%.
Beverage of food with required particle size distribution was prepared by emulsification in a certain weight percentage of inactivated Kluyveromyces cell dry powder prepared in Example 5, coconut oil, and water. The specific steps are as follows:
The results of the percentage content by weight of inactivated Kluyveromyces cell dry powder, coconut oil and water, the particle size distribution of composition, and the results of stability test are shown in Table 14 (A is defined as no demulsification and no stratification at all; B is defined as slight demulsification and a little amount of stratification at the top of the product; C is defined as obvious demulsification and a complete oil and water stratification of product):
Sauce of food with required particle size distribution was prepared by emulsification in a certain weight percentage of inactivated Kluyveromyces cell dry powder prepared in Example 5, coconut oil, and water. The specific steps are as follows:
The results of the percentage content by weight of inactivated Kluyveromyces cell dry powder, coconut oil and water, the particle size distribution of composition, and the results of stability test are shown in Table 15 (A is defined as no demulsification and no stratification at all; B is defined as slight demulsification and a little amount of stratification at the top of the product; C is defined as obvious demulsification and a complete oil and water stratification of product):
Beverage of food with required particle size distribution was prepared by emulsification in a certain weight percentage of inactivated Kluyveromyces cell dry powder prepared in Example 5, high oleic sunflower seed oil, and water. The specific steps are as follows:
The results of the percentage content by weight of inactivated Kluyveromyces cell dry powder, high oleic sunflower seed oil and water, the particle size distribution of composition, and the results of stability test are shown in Table 16 (A is defined as no demulsification and no stratification at all; B is defined as slight demulsification and a little amount of stratification at the top of the product; C is defined as obvious demulsification and a complete oil and water stratification of product):
Sauce of food with required particle size distribution was prepared by emulsification in a certain weight percentage of inactivated Kluyveromyces cell dry powder prepared in Example 5, high oleic sunflower seed oil, and water. The specific steps are as follows:
The results of the percentage content by weight of inactivated Kluyveromyces cell dry powder, high oleic sunflower seed oil and water, the particle size distribution of composition, and the results of stability test are shown in Table 17 (A is defined as no demulsification and no stratification at all; B is defined as slight demulsification and a little amount of stratification at the top of the product; C is defined as obvious demulsification and a complete oil and water stratification of product):
Beverage of food with required particle size distribution was prepared by emulsification in a certain weight percentage of inactivated Kluyveromyces cell dry powder prepared in Example 5, canola oil, and water. The specific steps are as follows:
The results of the percentage content by weight of inactivated Kluyveromyces cell dry powder, canola oil and water, the particle size distribution of composition, and the results of the stability test are shown in Table 18 (A is defined as no demulsification and no stratification at all; B is defined as slight demulsification and a little amount of stratification at the top of the product; C is defined as obvious demulsification and a complete oil and water stratification of product):
Sauce of food with required particle size distribution was prepared by emulsification in a certain weight percentage of inactivated Kluyveromyces cell dry powder prepared in Example 5, canola oil, and water. The specific steps are as follows:
The results of the percentage content by weight of inactivated Kluyveromyces cell dry powder, canola oil and water, the particle size distribution of composition, and the results of the stability test are shown in Table 19 (A is defined as no demulsification and no stratification at all; B is defined as slight demulsification and a little amount of stratification at the top of the product; C is defined as obvious demulsification and a complete oil and water stratification of product):
Beverage of food with required particle size distribution was prepared by emulsification in a certain weight percentage of inactivated Kluyveromyces cell dry powder prepared in Example 5, medium chain triglyceride, and water. The specific steps are as follows:
The results of the percentage content by weight of inactivated Kluyveromyces cell dry powder, medium chain triglyceride and water, the particle size distribution of composition, and the results of the stability test are shown in Table 20 (A is defined as no demulsification and no stratification at all; B is defined as slight demulsification and a little amount of stratification at the top of the product; C is defined as obvious demulsification and a complete oil and water stratification of product):
Sauce of food with required particle size distribution was prepared by emulsification in a certain weight percentage of inactivated Kluyveromyces cell dry powder prepared in Example 5, medium chain triglyceride, and water. The specific steps are as follows:
The results of the percentage content by weight of inactivated Kluyveromyces cell dry powder, medium chain triglyceride and water, the particle size distribution of composition, and the results of the stability test are shown in Table 21 (A is defined as no demulsification and no stratification at all; B is defined as slight demulsification and a little amount of stratification at the top of the product; C is defined as obvious demulsification and a complete oil and water stratification of product):
As results shown in Table 7, 14, 16, 18 and 20, beverages of food in the present disclosure were prepared by inactivated Kluyveromyces marxianus cells with rapeseed oil, coconut oil, high oleic sunflower seed oil, canola oil, and medium chain triglyceride, according to the proportion in the present disclosure, respectively. After shear emulsifying process until the particle sizes of each component in the suspension are all less than or equal to 100 μm (the frequency of particle size≤100 mμm is 100%), it is proved that all can maintain a stable and uniform status without stratification for at least 7 days at room temperature. It can even maintain a stable and uniform status without stratification for 28 days according to the preceding preferred proportion of beverages in the present disclosure (2.5-25% of inactivated Kluyveromyces cell powder, 1%-30% of edible oil).
As results shown in Table 9, 15, 17, 19 and 21, sauces of food in the present disclosure were prepared by inactivated Kluyveromyces marxianus cells with rapeseed oil, coconut oil, high oleic sunflower seed oil, canola oil, and medium chain triglyceride, according to the proportion in the present disclosure, respectively. After shear emulsifying process until the particle sizes of each component in the suspension are all less than or equal to 100 μm (the frequency of particle size≤100 mμm is 100%), it is proved that it can maintain a stable and uniform status without stratification for 28 days at room temperature.
Specifically, the edible oil is all common edible oil of different sources and nutrients that used in food industry, whose market price is at 10-300/L (canola oil is 10 CNY/L, high oleic acid sunflower seed oil is 16 CNY/L, coconut oil is 50 CNY/L, medium chain triglyceride is 300 CNY/L). The raw materials of food compositions in the present disclosure are cheap and have relatively low cost.
Understandably, Example 11-18 are exemplary embodiments. Except for inactivated Kluyveromyces marxianus cells, beverages or sauces of food had the same or similar stability during shelf life, which were prepared according to proportion in the present disclosure by the four other inactivated Kluyveromyces cells (Kluyveromyces hubeiensis, Kluyveromyces wickerhamii, Kluyveromyces thermotolerans, and Kluyveromyces lactis) in the present disclosure and edible oils other than rapeseed oil.
Kluyveromyces beverage was made of inactivated Kluyveromyces marxianus cell dry powder prepared in Example 5 according to shear emulsifying. Under the same method and proportion, the food composition was replaced by pea protein and sucrose fatty acid ester to obtain plant-based beverage and emulsifier beverage. The composition of beverages is shown in Table 22. The statuses of stratification and demulsification of the preceding three beverages were recorded on the on the 1st, 3rd, 7th, 14th and 28th day after the preparation of beverages (A is defined as no demulsification and no stratification at all; B is defined as slight demulsification and a little amount of stratification at the top of the product; C is defined as obvious demulsification and a complete oil and water stratification of product). The observation results are shown in Table 23 below.
Kluyveromyces
marxianus cell dry
Kluyveromyces
As shown in Table 23, the Kluyveromyces beverage prepared without exogenous food additives completely maintained a stable and uniform suspension status without stratification during the 28-day shelf life observation period, without any demulsification either; the plant-based beverage prepared without exogenous food additives showed obvious stratification and demulsification on the 7th day during the 28-day shelf life observation period; the emulsifier beverage prepared by sucrose fatty acid ester which is an emulsifying food additive showed a little stratification and slight demulsification on the 7th day, and obvious stratification and demulsification on the 28th day during the 28-day shelf life observation period. It can be seen that Kluyveromyces beverage has the value of developing microbic beverages and microbic food raw materials.
The preceding descriptions of specific exemplary embodiments are for the purpose of explanation and illustration. These descriptions are not intended to limit the present disclosure to the precise form disclosed, and it is clear that many changes and variations can be made according to the preceding instructions. The purpose of selection and description of exemplary embodiments is to explain the specific principles and their practical applications of the present disclosure so that those skilled in the art can implement and utilize various exemplary embodiments and different options and changes of the present disclosure. The scope of the present disclosure is intended to be limited by the claims and their equivalents.
| Number | Date | Country | Kind |
|---|---|---|---|
| 202111359573.X | Nov 2021 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2022/131380 | 11/11/2022 | WO |
