The present invention relates to a method of producing metal nanoparticles and metal sulfide nanoparticles using a recombinant microorganism, and more particularly to a method of producing metal nanoparticles and metal sulfide nanoparticles using a recombinant microorganism co-expressing metallothionein and phytochelatin synthase, which are heavy metal-adsorbing proteins.
As interest in nanotechnology has increased, various studies have been actively conducted to develop nanometer-sized new materials in various fields, including as physics, chemistry, materials, electricity, and electronics. In particular, materials comprising metal nanostructures can be applied to the fabrication of high-efficiency electronic, optics, photoelectronics, electronic devices, bioactive molecule detection devices, and catalysts, which were difficult to realize by bulk materials made using existing technologies, and these materials can also be applied in medicines, cosmetics, energy conversion and storage, etc., indicating that these materials are used in various and broad applications (Shedbalkar U et al., Adv Colloid Interface Sci, 209:40-48, 2014; Fan W et al., Phys Chem Chem Phys, 15(8):2632-2649, 2013; Korbekandi H et al., Crit Rev Biotechnol 29(4):279-306, 2009). Thus, studies have been actively conducted to synthesize novel metal nanoparticles or to discover the novel properties of existing metal nanoparticles.
In the production of conventional metal nanoparticles, a chemical method of synthesizing the metal nanoparticles from a metal element using a chemical reducing agent has been mainly used. However, this method has disadvantages in that it requires a toxic organic solvent and a costly catalyst and requires high temperature and high pressure conditions, and thus it causes environmental pollution problems and has low energy efficiency and economic efficiency. For this reason, as an environmentally friendly and cost-effective alternative, biological methods using organisms having reductase have been proposed. In fact, cases have been reported in which metal nanoparticles are synthesized from metal elements using microorganisms, enzymes, plants, or the like (Paza G A et al., Int J Mol Sci 15(8):13720-13737, 2014; Hulkoti N I et al., Colloids Surf B Biointerfaces, 121:474-483, 2014; Moghaddam et al., Molecules, 20(9):16540-16565, 2015; Park T J et al., Appl Microbiol Biotechnol, 1-14, 2015). However, such methods use the biological mechanisms of organisms themselves without changes, and have a limitation in that elements that are applied for the synthesis of nanoparticles are limited to gold, silver, copper, cadmium, iron, selenium and the like (Quester K et al., Micron, 54-55:1-27, 2013; Bharde A et al., J Nanosci Nanotechnol, 7(12):4369-4377. 2007; Shedbalkar U et al., Adv Colloid Interface Sci, 209:40-48, 2014; Sriram M I et al., Methods Mol Biol, 906:33-43, 2012; Gurunathan S et al., Colloids Surf B Biointerfaces, 74(1):328-335, 2009).
Previously, the present inventors produced metal nanoparticles from elements such as zinc (Zn), selenium (Se), tellurium (Te), cesium (Cs), copper (Cu), lead (Pb), nickel (Ni), manganese (Mn), mercury (Hg), cobalt (Co) and chromium (Cr) by use of recombinant microorganisms that express heavy metal-adsorbing protein (Lee et al., ACS Nano 6(8):6998-7008, 2012; Korean Patent No. 10-0755746). However, there were limitations in producing metal nanoparticles using various other metal elements, and there was a problem in that the yield is also low.
Meanwhile, metal sulfide nanoparticles are attracting attention as next-generation electrode materials and energy storage materials because of their unique and excellent characteristics (Rui X et al., Nanoscale, 6(17): 9889-9924, 2014). Accordingly, prior art technologies related to methods for producing metal sulfide nanoparticles have been reported, including a method of producing metal sulfide nanocrystals using a thiol compound as a sulfur precursor, which is related to a method of obtaining metal sulfide crystals by reacting a metal precursor with a thiol compound (Korean Patent No. 10-0621309); plate-like or linear indium composite nanomaterials for use as a buffer in a solar cell (Korean Patent No. 10-1305554); a method of producing tin sulfide nanoparticles by heating tin sulfide precursors and a method of fabricating a lithium ion battery using the same (Korean Patent No. 10-0896656), and the like. However, the above methods have disadvantages in that they require a high reaction temperature of 300° C. or higher, are not environmentally friendly due to the use of organic solvents such as toluene, and require a subsequent heat treatment process, which is disadvantageous in terms of production costs. In addition, CdS and PbS (Joo J et al., J. Am. Chem. Soc, 125(36):11100-11105, 2003), which are used as semiconductor materials, have negative environmental impacts, and thus it is required to synthesize metal sulfide nanoparticles using elements other than cadmium- or lead-based metal sulfides.
Accordingly, the present inventors have made extensive efforts to produce various metal nanoparticles and metal sulfide nanoparticles which have not been synthesized in conventional art. As a result, the present inventors have found that, when culture conditions are optimized using a recombinant microorganism co-expressing metallothionein and phytochelatin synthase, which are heavy metal-adsorbing proteins, metal nanoparticles and metal sulfide nanoparticles can be produced from elements which have not been reported as elements for synthesis of metal nanoparticles in conventional art, and the yield of metal nanoparticles which have been produced by conventional methods is also significantly increased, thereby completing the present invention.
The information disclosed in the Background Art section is only for the enhancement of understanding of the background of the present invention, and therefore may not contain information that forms a prior art that would already be known to a person of ordinary skill in the art.
It is an object of the present invention to provide a method of producing single-element metal nanoparticles using a recombinant microorganism.
Another object of the present invention is to provide a method of producing metal alloy nanoparticles using a recombinant microorganism.
Still another object of the present invention is to provide a method of producing metal sulfide nanoparticles using a recombinant microorganism.
Yet another object of the present invention is to provide the use of the produced metal nanoparticles.
To achieve the above object, the present invention provides a method for producing a single-element metal nanoparticle, comprising the steps of: (a) culturing a recombinant microorganism into which a metallothionein-encoding gene and a phytochelatin synthase-encoding gene are introduced; (b) adding to a medium of step (a) a metal ion selected from the group consisting of zinc (Zn), selenium (Se), tellurium (Te), cesium (Cs), copper (Cu), lead (Pb), nickel (Ni), manganese (Mn), mercury (Hg), cobalt (Co), chromium (Cr), cadmium (Cd), strontium (Sr), iron (Fe), gold (Au), silver (Ag), praseodymium (Pr), gadolinium (Gd), barium (Ba), zirconium (Zr), molybdenum (Mo), indium (In), tin (Sn), lanthanum (La), cerium (Ce), neodymium (Nd), samarium (Sm), yttrium (Y), aluminum (Al), and europium (Eu), followed by additional culturing, thereby producing a single-element metal nanoparticle; and (c) recovering the produced single-element metal nanoparticle.
The present invention also provides a method for producing a metal alloy nanoparticle, comprising the steps of: (a) culturing a recombinant microorganism into which a metallothionein-encoding gene and a phytochelatin synthase-encoding gene are introduced; (b) adding to a medium of step (a) two or more metal ions selected from the group consisting of zinc (Zn), selenium (Se), tellurium (Te), cesium (Cs), copper (Cu), lead (Pb), nickel (Ni), manganese (Mn), mercury (Hg), cobalt (Co), chromium (Cr), cadmium (Cd), strontium (Sr), iron (Fe), gold (Au), silver (Ag), praseodymium (Pr), gadolinium (Gd), barium (Ba), zirconium (Zr), molybdenum (Mo), indium (In), tin (Sn), lanthanum (La), cerium (Ce), neodymium (Nd), samarium (Sm), yttrium (Y), aluminum (Al), and europium (Eu), followed by additional culturing, thereby producing a metal alloy nanoparticle; and (c) recovering the produced metal alloy nanoparticle.
The present invention also provides a method for producing a metal sulfide nanoparticle, comprising the steps of: (a) culturing a recombinant microorganism into which a metallothionein-encoding gene and a phytochelatin synthase-encoding gene are introduced; (b) adding to a medium of step (a) a metal ion selected from the group consisting of i) zinc (Zn), selenium (Se), tellurium (Te), cesium (Cs), copper (Cu), lead (Pb), nickel (Ni), manganese (Mn), mercury (Hg), cobalt (Co), chromium (Cr), cadmium (Cd), strontium (Sr), iron (Fe), gold (Au), silver (Ag), praseodymium (Pr), gadolinium (Gd), barium (Ba), zirconium (Zr), molybdenum (Mo), indium (In), tin (Sn), lanthanum (La), cerium (Ce), neodymium (Nd), samarium (Sm), yttrium (Y), aluminum (Al), and europium (Eu); and ii) sulfur, followed by additional culturing, thereby producing a metal sulfide nanoparticle; and (c) recovering the produced metal sulfide nanoparticle.
The present invention also provides a contrast agent comprising a silver tellurite (Ag2TeO3) nanoparticle.
The present invention also provides an electrode comprising a silver tellurite (Ag2TeO3) nanoparticle.
Unless defined otherwise, all the technical and scientific terms used herein have the same meaning as those generally understood by one of ordinary skill in the art to which the invention pertains. Generally, the nomenclature used herein and the experiment methods, which will be described below, are those well known and commonly employed in the art.
In the present invention, in order to synthesize various metal nanoparticles using a biological method, a recombinant microorganism co-expressing two heavy metal-adsorbing proteins (metallothionein and phytochelatin synthase) was cultured in media containing various metal elements. As a result, it was found that single-element metal nanoparticles were synthesized.
In one example of the present invention, a recombinant microorganism was cultured, and then additionally cultured in medium containing a single-element metal ion such as barium (Ba), zirconium (Zr), molybdenum (Mo), indium (In), tin (Sn), lanthanum (La), cerium (Ce) or praseodymium (Pr), and as a result, it could be seen that amorphous single nanoparticles were synthesized. Meanwhile, when the recombinant microorganism was cultured in medium containing cobalt (Co), nickel (Ni), zinc (Zn) or cadmium (Cd) ions, it was observed that single-element metal nanoparticles were not synthesized. However, when the recombinant microorganism was cultured in media containing these metal ions after increasing the pH of the media, it could be seen that crystalline metal particles of cobalt oxide (Co3O4), nickel hydroxide (Ni(OH)2), zinc peroxide (ZnO) or cadmium hydroxide (Cd(OH)2) were produced. In addition, when the recombinant microorganism was cultured in medium containing lanthanum (La), cerium (Ce), praseodymium (Pr), neodymium (Nd), samarium (Sm), europium (Eu) or gadolinium (Gd) ions, it was observed that amorphous single-element metal nanoparticles were synthesized. However, when the recombinant microorganism was cultured in media containing the above-described metal ions after increasing the pH of the media, it was observed that crystalline single-element metal nanoparticles of lanthanum hydroxide (La(OH)3), cerium oxide (CeO2), praseodymium hydroxide (Pr(OH)3), neodymium hydroxide (Nd(OH)3), samarium hydroxide (Sm(OH)3), europium hydroxide (Eu(OH)3) or gadolinium hydroxide (Gd(OH)3) were produced.
Therefore, in a first aspect, the present invention is directed to a method for producing single-element metal nanoparticles, comprising the steps of: (a) culturing in a medium a recombinant microorganism having introduced therein a metallothionein-encoding gene and a phytochelatin synthase-encoding gene; (b) adding to the medium of step (a) a metal ion selected from the group consisting of zinc (Zn), selenium (Se), tellurium (Te), cesium (Cs), copper (Cu), lead (Pb), nickel (Ni), manganese (Mn), mercury (Hg), cobalt (Co), chromium (Cr), cadmium (Cd), strontium (Sr), iron (Fe), gold (Au), silver (Ag), praseodymium (Pr), gadolinium (Gd), barium (Ba), zirconium (Zr), molybdenum (Mo), indium (In), tin (Sn), lanthanum (La), cerium (Ce), neodymium (Nd), samarium (Sm), yttrium (Y), aluminum (Al), and europium (Eu), followed by additional culturing, thereby producing single-element metal nanoparticles; and (c) recovering the produced single-element metal nanoparticles.
In the present invention, the added metal ion may be selected from the group consisting of zinc (Zn), selenium (Se), tellurium (Te), cesium (Cs), copper (Cu), lead (Pb), nickel (Ni), manganese (Mn), mercury (Hg), cobalt (Co), chromium (Cr), cadmium (Cd), strontium (Sr), iron (Fe), gold (Au), silver (Ag), praseodymium (Pr), gadolinium (Gd), barium (Ba), zirconium (Zr), molybdenum (Mo), indium (In), tin (Sn), lanthanum (La), cerium (Ce), neodymium (Nd), samarium (Sm), yttrium (Y), aluminum (Al), and europium (Eu), but is not limited thereto.
In the present invention, the produced metal nanoparticles may be selected from the group consisting of cobalt oxide (Co3O4), nickel hydroxide (Ni(OH)2), zinc peroxide (ZnO), cadmium hydroxide (Cd(OH)2), cobalt hydroxide (Co(OH)2), barium carbonate (BaCO3), lanthanum hydroxide (La(OH)3), cerium oxide (CeO2), praseodymium hydroxide (Pr(OH)3), neodymium hydroxide (Nd(OH)3), samarium hydroxide (Sm(OH)3), europium hydroxide (Eu(OH)3), cerium hydroxide (Ce(OH)3), yttrium hydroxide (Y(OH)3), aluminum hydroxide (Al(OH)3), and gadolinium hydroxide (Gd(OH)3), but are not limited thereto.
In the present invention, it could be found that the surface functional groups of the single-element metal nanoparticles comprise 3300-3000 cm−1 (OH groups), 2960-2850 cm−1 (C—H), 1650-1660 cm−1 (amide I), 1540-1535 cm−1 (amide II), 1240-1234 cm−1 (amide III), and 1150-1030 cm−1 (C═O) in an IR spectrum range of 400 to 4000 cm−1.
In the present invention, when the culture in step (b) was performed after adjusting the initial pH to 7.3 to 7.7, it could be seen that the metal nanoparticles were produced with the highest efficiency while the pH increased to 8.0 to 9.0 with the passage of time. In addition, it could be seen that the potential (Eh) of the medium when the metal nanoparticles were produced with the highest efficiency was −0.5 V to +0.5 V.
Thus, in the present invention, the pH of the medium in step (b) may be adjusted so that it reaches an optimal pH of 8.0-9.0 at which the single-element metal nanoparticles are produced by additional culture of the recombinant microorganism.
To this end, the initial pH of the medium in step (b) before additional culture is preferably increased to 7.3 to 7.7, more preferably 7.4 to 7.6.
In the present invention, the initial pH of the medium in step (a) before culture may be 6 to 7, preferably 6.4 to 6.6.
When the pH in the microorganism culture step and the pH in the metal nanoparticle production step are adjusted as described above, it is possible to synthesize metal nanoparticles which have been difficult to synthesize in conventional art. Metal nanoparticles that can be synthesized by increasing the pH may be selected from the group consisting of cobalt oxide (Co3O4), nickel hydroxide (Ni(OH)2), zinc peroxide (ZnO) and cadmium hydroxide (Cd(OH)2), but are not limited thereto.
In addition, the single-element metal nanoparticles including cobalt oxide (Co3O4), nickel hydroxide (Ni(OH)2), zinc peroxide (ZnO), cadmium hydroxide (Cd(OH)2), lanthanum hydroxide (La(OH)3), cerium oxide (CeO2), praseodymium hydroxide (Pr(OH)3), neodymium hydroxide (Nd(OH)3), samarium hydroxide (Sm(OH)3), europium hydroxide (Eu(OH)3), and gadolinium hydroxide (Gd(OH)3) were amorphous when the pH was not adjusted, but crystalline nanoparticles were produced as the pH was increased.
In another example of the present invention, the recombinant microorganism was cultured, and then it was observed that i) metal alloy nanoparticles of cobalt iron oxide (CoFe2O4) were produced in a medium containing cobalt and iron (Co, CoCl2, 0.5 mM, Fe, Fe(NO)3.6H2O, 0.5 mM); ii) metal alloy nanoparticles of nickel iron oxide (NiFe2O4) were produced in a medium containing nickel and iron (Ni, NiCl2.6H2O, 0.5 mM, Fe, Fe(NO)3.6H2O, 0.5 mM); iii) metal alloy nanoparticles of zinc manganese oxide (ZnMn2O4) were produced in a medium containing zinc and manganese (Zn, ZnSO4.7H2O, 0.5 mM, MnCl24H2O); iv) metal alloy nanoparticles of zinc iron oxide (ZnFe2O4) were produced in a medium containing zinc and iron ions (Zn, ZnSO4.7H2O, 0.5 mM, Fe, Fe(NO)3.6H2O, 0.5 mM); and v) metal alloy nanoparticles of silver tellurite (Ag2TeO3) were produced in a medium containing silver (Ag, AgNO3, 0.25, 0.5 mM) and tellurium (Te, NaTeO3, 0.25, 0.5 mM) ions.
Therefore, in a second aspect, the present invention is directed to a method for producing metal alloy nanoparticles, comprising the steps of: (a) culturing in a medium a recombinant microorganism having introduced therein a metallothionein-encoding gene and a phytochelatin synthase-encoding gene; (b) adding to the medium of step (a) two or more metal ions selected from the group consisting of zinc (Zn), selenium (Se), tellurium (Te), cesium (Cs), copper (Cu), lead (Pb), nickel (Ni), manganese (Mn), mercury (Hg), cobalt (Co), chromium (Cr), cadmium (Cd), strontium (Sr), iron (Fe), gold (Au), silver (Ag), praseodymium (Pr), gadolinium (Gd), barium (Ba), zirconium (Zr), molybdenum (Mo), indium (In), tin (Sn), lanthanum (La), cerium (Ce), neodymium (Nd), samarium (Sm), yttrium (Y), aluminum (Al), and europium (Eu), followed by additional culturing, thereby producing metal alloy nanoparticles; and (c) recovering the produced metal alloy nanoparticles.
In the present invention, the added metal ions may be two or more metal ions selected from the group consisting of zinc (Zn), selenium (Se), tellurium (Te), cesium (Cs), copper (Cu), lead (Pb), nickel (Ni), manganese (Mn), mercury (Hg), cobalt (Co), chromium (Cr), cadmium (Cd), strontium (Sr), iron (Fe), gold (Au), silver (Ag), praseodymium (Pr), gadolinium (Gd), barium (Ba), zirconium (Zr), molybdenum (Mo), indium (In), tin (Sn), lanthanum (La), cerium (Ce), neodymium (Nd), samarium (Sm), yttrium (Y), aluminum (Al), and europium (Eu), but is not limited thereto.
In the present invention, it was found that the metal alloy nanoparticles produced by the above-described method consist of zinc manganese oxide (ZnMn2O4), zinc iron oxide (ZnFe2O4), nickel iron oxide (NiFe2O4), cobalt iron oxide (CoFe2O4), cadmium selenide (CdSe), cadmium zinc (CdZn), cadmium telluride (CdTe), zinc selenide (SeZn), cadmium selenide zinc (CdSeZn), strontium gadolinium (SrGd), praseodymium gadolinium (PrGd), cadmium cesium CdCs), cobalt iron (FeCo), iron cobalt nickel compound (FeCoNi), and silver tellurite (Ag2TeO3).
In the present invention, it could be found that the surface functional groups of the metal alloy nanoparticles comprise 3300-3000 cm−1 (O—H), 2960-2850 cm−1 (C—H), 1650-1660 cm−1 (amide I), 1540-1535 cm−1 (amide II), 1240-1234 cm−1 (amide III), and 1150-1030 cm−1 (C═O) in an IR spectrum range of 400 to 4000 cm−1.
It could be seen that, among the metal nanoparticles synthesized by the above-described method, cobalt iron oxide (CoFe2O4) nanoparticles are ferromagnetic, and nickel iron oxide (NiFe2O4), zinc manganese oxide (ZnMn2O4) and zinc iron oxide (ZnFe2O4) nanoparticles are paramagnetic.
In particular, in the second aspect, it could be seen that when silver (Ag) and tellurium (Te) are added simultaneously, silver tellurite (Ag2TeO3) nanoparticles were synthetized, and the surface functional groups of the silver tellurite (Ag2TeO3) metal alloy nanoparticles comprise 3300-3000 cm−1 (O—H), 2960-2850 cm−1 (C—H), 1650-1660 cm−1 (amide I), 1540-1535 cm−1 (amide II), 1240-1234 cm−1 (amide III), and 1150-1030 cm−1 (C═O).
Therefore, in a third aspect, the present invention is directed to the novel use of silver tellurite (Ag2TeO3) nanoparticles synthesized using a recombinant microorganism co-expressing metallothionein and phytochelatin synthase.
The silver tellurite (Ag2TeO3) nanoparticles are diamagnetic, and thus can be used as a contrast agent, wherein the contrast agent may be used for imaging selected from the group consisting of optical imaging, magnetic resonance imaging, precision nuclear medicine imaging, and ultrasound imaging, but is not limited thereto.
Furthermore, the silver tellurite (Ag2TeO3) nanoparticles show different CV curves depending on scan rate, and have electrochemical characteristics showing peaks at about 0.30 V and 0.40 V in an anode and peaks at −0.50 V and −0.20 V in a cathode. Thus, the silver tellurite (Ag2TeO3) nanoparticles may be used as a component of an electrode, wherein the electrode may be used in a cell selected from the group consisting of lithium ion cells, fuel cells, solar cells, hydrogen cells, and secondary cells, but is not limited thereto. For example, the metal nanoparticles according to the present invention may be used in an anode for a lithium-ion cell, particularly, a lithium-ion secondary cell. In addition, the metal nanoparticles according to the present invention may be used in an anode for a lithium-ion cell, particularly a lithium-ion secondary cell, which comprises an electrically active material.
The silver tellurite (Ag2TeO3) nanoparticles according to the present invention may be used as a component of a biosensor and for a semiconductor device.
In addition, the metal nanoparticles synthesized according to the present invention may be used in various applications based on their characteristics or activity. Examples of applications in which the metal nanoparticles according to the present invention can be used include, but are not limited to, electrode activation, a catalyst for glucose oxidation, a catalyst for carbon monoxide oxidation, a catalyst for carbon compound oxidation, a catalyst for synthesis of pyrano[2,3-d] pyrimidines, a catalyst for cobalt compound synthesis, a catalyst for iron compound synthesis, a composition for a storage battery, a composition for a battery, a composition for a lithium secondary cell, a photoelectronic device, a semiconductor device, a light-emitting diode, a self-cooling device, a carrier for drug delivery, a radiotherapy enhancer, an MRI contrast agent, a composition for medical diagnosis, a composition for high-density magnetic recording medium, a composition for electromagnetic wave absorption, an antibacterial composition, a composition for treatment/prevention of oral diseases, a composition for IR detection, a composition for X-ray detection, a composition for gamma-ray detection, a magnetic refrigerant composition, a raw material for a permanent magnet, a biosensor, a composition for graphene tubes, a food additive composition, a tobacco filter composition, a paint pigment composition, a paint drying composition, a porcelain colorant composition, and the like.
In addition, the metal nanoparticles according to the present invention may be used in any fields in which the development and proliferation of microorganisms must be suppressed. Specifically, the metal nanoparticles are advantageously used in medical devices, hand rails, door handles, mobile phones, keyboards, etc. Furthermore, the metal nanoparticles according to the present invention are useful for the disinfection and general antimicrobial treatment, such as deodorizing, of the skin, mucous membrane and hair, preferably for the disinfection of hands and wounds, and may be utilized as an antimicrobial agent in various product forms or compositions for personal and household care use or for industrial and hospital applications including, but not limited to, cosmetic compositions for skin and hair care, for example, lotions; creams; oils; gels; powders; wipes; deodorants like sprays, sticks and roll-ons; cleansers like shower gels; bath additives; liquid and solid soaps (based on synthetic surfactants and salts of saturated and/or unsaturated fatty acids); aqueous and/or alcoholic solutions, e.g., cleansing solutions for the skin; moist cleaning cloths; hand sanitizers; shampoos; rinses; etc.; oral hygiene compositions, for example, in the form of a gel, a paste, a cream or an aqueous preparation (mouthwash); hard surface cleaners, e.g., disinfectant sprays, liquids, or powders; dish or laundry detergents (liquid or solid), floor waxes, glass cleaners, etc.; and industrial and hospital applications (e.g., sterilization of instruments, medical devices, gloves; contact lenses, contact lens cases, contact lens storage solutions, contact lens cleaning solutions).
In another example of the present invention, it was found that the recombinant microorganism was additionally cultured for 12 hours in the medium supplemented with sulfur (S, Na2S, 1 mM) together with silver (Ag, AgNO3, 0.5 mM), indium (In, InCl24H2O, 0.5 mM)), manganese (Mn, MnSO45H2O, 0.5 mM) or tin (Sn, SnCl24H2O, 0.5 mM), and as a result, it was shown that silver sulfide, indium sulfide, manganese sulfide or tin sulfide nanoparticles were produced.
Therefore, in a fourth aspect, the present invention is directed to a method for producing metal sulfide nanoparticles, comprising the steps of: (a) culturing in a medium a recombinant microorganism having introduced therein a metallothionein-encoding gene and a phytochelatin synthase-encoding gene; (b) adding to the medium of step (a) a metal ion selected from the group consisting of i) zinc (Zn), selenium (Se), tellurium (Te), cesium (Cs), copper (Cu), lead (Pb), nickel (Ni), manganese (Mn), mercury (Hg), cobalt (Co), chromium (Cr), cadmium (Cd), strontium (Sr), iron (Fe), gold (Au), silver (Ag), praseodymium (Pr), gadolinium (Gd), barium (Ba), zirconium (Zr), molybdenum (Mo), indium (In), tin (Sn), lanthanum (La), cerium (Ce), neodymium (Nd), samarium (Sm), and europium (Eu); and ii) sulfur, followed by additional culturing, thereby producing metal sulfide nanoparticles; and (c) recovering the produced metal sulfide nanoparticles.
In the present invention, the added metal ion may be selected from the group consisting of zinc (Zn), selenium (Se), tellurium (Te), cesium (Cs), copper (Cu), lead (Pb), nickel (Ni), manganese (Mn), mercury (Hg), cobalt (Co), chromium (Cr), cadmium (Cd), strontium (Sr), iron (Fe), gold (Au), silver (Ag), praseodymium (Pr), gadolinium (Gd), barium (Ba), zirconium (Zr), molybdenum (Mo), indium (In), tin (Sn), lanthanum (La), cerium (Ce), neodymium (Nd), samarium (Sm), and europium (Eu), but is not limited thereto.
In the present invention, the produced metal sulfide nanoparticles may be selected from the group consisting of silver sulfide (Ag2S), indium sulfide (InS), manganese sulfide (MnS), zinc sulfide (ZnS), copper sulfide (CuS), cadmium sulfide (CdS), gold sulfide (AuS), nickel sulfide (NiS), cobalt sulfide (CoS), mercury sulfide (HgS), and tin sulfide (SnS), but is not limited thereto.
In the present invention, it could be found that the surface functional groups of the metal sulfide nanoparticles comprise 3300-3000 cm−1 (O—H), 2960-2850 cm−1 (C—H), 1650-1660 cm−1 (amide I), 1540-1535 cm−1 (amide II), 1240-1234 cm−1 (amide III), and 1150-1030 cm−1 (C═O) in an IR spectrum range of 400 to 4000 cm−1.
In the first, second, and fourth aspects of the present invention, the microorganism may be selected from the group consisting of bacteria, yeasts, algae, archaea, and fungi, but is not limited thereto.
In the first, second, and fourth aspects of the present invention, the metallothionein-encoding gene may be represented by SEQ ID NO: 1, and the phytochelatin synthase-encoding gene is represented by SEQ ID NO: 2, but they are not limited thereto.
In the present invention, metal nanoparticles produced in the recombinant microorganism can be recovered by disrupting the microorganism, removing the cell debris by filtration, and collecting the metal nanoparticles from the filtered solution. In one embodiment, the metal nanoparticles can be recovered by centrifuging the microbial culture at 3500 rpm and 4° C. for 15 minutes, removing the supernatant, collecting the microbial pellet, washing the collected microbial pellet three times with PBS buffer, disrupting the cell suspension with a sonicator (VCX-600, Sonics and Materials Inc., USA) equipped with an ultrasonic probe (40 T, Sonics and Materials Inc., USA) (on for 3 sec and off for 3 sec), removing the cell debris by filtration through a 8-μm pore size cellulose filter and then through a 0.1-μm pore size membrane filter (Whatman), and collecting the nanoparticles from the filtered solution. In addition to this method, any method known in the art may also be used to recover the nanoparticles from the microbial cells or the cell culture medium.
In examples of the present invention, a specific medium and culture method has been illustrated, but it will be obvious to a person of ordinary skill in the art that other media can be used as reported in literatures. In examples of the present invention, E. coli was used as a microorganism, it will be obvious to those skilled in the art that other bacteria, yeasts and fungi could be used. In addition, although the following examples have illustrated only a specific strain-derived gene as a gene to be introduced, it will be obvious to those skilled in the art that any gene can be used as the gene to be introduced without limitations, as long as it is expressed in a host cell being introduced to show the same activity as that of the above gene.
In the present invention, the concentration of metal ions may be 5 mM to 0.01 mM, preferably 3 mM to 0.1 mM, more preferably 2 to 0.5 mM. If the concentration of metal ions is more than 5 mM, their toxicity to cells may increase to inhibit the growth of the cells, as reported in the literatures (Xiu Z M et al., Nano Lett., 12(8): 4271-4275, 2012; Nies D H, Appl Microbiol Biotechnol, 51(6):730-750, 1999). Meanwhile, the size of metal nanoparticles can be controlled by controlling the concentration of metal ions.
In the present invention, an aqueous metal compound can be used as a metal element. For example, an aqueous metal salt, an aqueous metal oxide salt, or a combination thereof can be used.
As used herein, the term “plasmid” means a DNA construct containing a DNA sequence operably linked to a suitable control sequence capable of effecting the expression of the DNA in a suitable host. The plasmid may be a vector, a phage particle, or simply a potential genomic insert. Once incorporated into a suitable host, the plasmid may replicate and function independently of the host genome, or may in some instances, integrate into the genome itself. In the present specification, “plasmid” and “vector” are sometimes used interchangeably, as the plasmid is the most commonly used form of vector. However, the present invention is intended to include other types of vectors with the same function as that would be known or known in the art.
As used herein, the term “recombinant microorganism” refers to a microorganism produced by modifying a part of a gene for a specific purpose, such as introducing a useful gene of any organism into another microorganism by using a genetic recombination technique, deleting a part of a gene of a host microorganism, or adjusting an expression amount. It will be apparent to those skilled in the art that the term can be replaced with the term “genetically modified microorganism”, “recombinant strain” or the like. The microorganism may include algae, bacteria, protozoa, fungi, yeast and the like. The microorganism can be preferably selected from the group consisting of bacteria, yeast, and fungi. More preferably, E. coli can be selected.
As used herein, the term “crystalline” means a state in which a certain solid forms a crystal in a repetitive atomic arrangement state.
As used herein, the term “amorphous” means an amorphous state, i.e., a state in which a certain solid does not form a crystal in a repetitive atomic arrangement state.
As used herein, the term “nanoparticle” means a particle having a diameter of a nanometer unit.
Hereinafter, the present invention will be described in further detail with reference to examples. It will be obvious to a person having ordinary skill in the art that these examples are for illustrative purposes only and are not to be construed to limit the scope of the present invention. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.
1-1: Construction of Metallothionein Expression Vector
Using the genomic DNA of Pseudomonas putida KT2440 as a template, polymerase chain reaction (PCR) was performed using primers of SEQ ID NOs: 3 and 4, thereby constructing an metallothionein (MT) gene fragment encoding the nucleotide sequence of metallothionein (MT).
Next, the constructed metallothionein fragment was subjected to agarose gel electrophoresis to isolate a 243-bp metallothionein gene fragment which was then digested with two restriction enzymes (EcoRI and PstI). Meanwhile, a pTac15K plasmid which is inducible and containing tac promoter was digested with two restriction enzymes (EcoRI and PstI), and then mixed and ligated with the MT fragment by T4 DNA ligase. The ligation product was transformed into E. coli DH5a by a heat shock method. The transformed strain was selected on LB medium containing kanamycin antibiotic (100 g/L), and a pYJ-MT recombinant plasmid was constructed therefrom (
1-2: Construction of Phytochelatin Synthase Expression Vector
In order to obtain phytochelatin synthase (PCS) from Arabidopsis thalinana, a phytochelatin synthase gene fragment encoding the nucleotide sequence of phytochelatin synthase was constructed by PCR using the complementary DNA (cDNA) of Arabidopsis thalinana as a template. The PCR was performed using primers of SEQ ID NOs: 5 and 6.
Next, the constructed phytochelatin synthase fragment was subjected to agarose gel electrophoresis to isolate a 1458-bp phytochelatin synthase gene fragment which was then digested with two restriction enzymes (EcoRI and SphI). Meanwhile, a pTac15K plasmid which is inducible and containing tac promoter was digested with two restriction enzymes (EcoRI and SphI), and then mixed and ligated with the phytochelatin synthase fragment by T4 DNA ligase. The ligation product was transformed into E. coli DH5a by a heat shock method. The transformed strain was selected on LB plate medium containing kanamycin antibiotic (100 g/L), and a pYJ-PCS recombinant plasmid was constructed therefrom (
1-3: Construction of Vector Co-Expressing Metallothionein and Phytochelatin Synthase
In order to construct a vector co-expressing metallothionein and phytochelatin synthase, pYJ-MT-PCS was constructed from the pYJ-MT and pYJ-PCS plasmids constructed as described above. Specifically, the pYJ-PCS vector was subjected to polymerase chain reaction (PCR) using primers of SEQ ID NOs: 7 and 8.
Then, a 1545-bp pYJ-PCS gene fragment was isolated from the PCR fragment by agarose gel electrophoresis. Meanwhile, the pYJ-MT plasmid was digested with two restriction enzymes (PstI and SphI), and then mixed and ligated with the pYJ-PCS fragment by T4 DNA ligase. The ligation product was transformed into E. coli DH5a by a heat shock method. The transformed strain was selected on LB plate medium containing kanamycin antibiotic (100 g/L), and a pYJ-MT-PCS recombinant plasmid was constructed therefrom (
1-4: Production of Transformed Recombinant Microorganism and Induction of Expression of Heavy Metal-Adsorbing Proteins
The recombinant plasmid pYJ-MT-PCS constructed in Example 1-3 so as to co-express metallothionein and phytochelatin synthase was introduced into E. coli DH5a. The transformed recombinant E. coli strain was inoculated into a 250 mL flask containing 100 mL of LB liquid medium and was cultured at 37° C. When the E. coli strain reached an optical density (OD) of 0.6 at a wavelength of 600 nm, IPTG (isopropyl-D-1-thiogalactopyranoside) was added to the medium to induce expression of metallothionein and phytochelatin synthase. The tac promoter inserted in the pYJ-MT-PCS was inducible by IPTG, and IPTG was added to the medium to a final concentration of 1 mM to induce the expression.
2-1: Synthesis of Single-Element Metal Nanoparticles Using Recombinant Microorganism
One hour after inducing expression of metallothionein and phytochelatin synthase in the recombinant microorganism produced in Example 1 above by IPTG, barium (Ba, (CH3COO)2Ba, 0.5 mM), zirconium (Zr, K2ZrF6, 0.5 mM), molybdenum (Mo, Na2MoO42H2O, 0.5 mM), indium (In, InCl24H2O, 0.5 mM), tin (Sn, SnCl24H2O, 0.5 mM), lanthanum (La, La(NO3)36H2O, 0.5 mM), cerium (Ce, Ce(NO3)36H2O, 0.5 mM) or praseodymium (Pr, Pr(NO3)36H2O, 0.5 mM) was added to the LB liquid medium (Tryptone 10 g/L, yeast extract 5 g/L, NaCl 5 g/L, pH 6.5) in which the recombinant E. coli strain has been cultured, after which the recombinant microorganism was additionally cultured for 12 hours. The culture was centrifuged at 3500 rpm and 4° C. for 15 minutes, after which the supernatant was discarded and the E. coli pellet was collected. The E. coli pellet was washed three times with PBS buffer, and then dried in a freeze-dryer under vacuum for one day or more. Synthesis of nanoparticles was examined using transmission electron microscopy (TEM) (Tecnai F20, Philips, Netherlands) and energy-dispersive X-ray spectroscopy (EDX) HD/MAX-2,500, Rigaku, Japan) with CuKa radiation (λ=1.5406 Å)).
As a result, as can be seen in
2-2: Analysis of Surface Functional Groups of Single-Element Metal Nanoparticles Synthesized Using Recombinant Microorganism
In order to confirm the surface functional groups of the single-element metal nanoparticles synthesized in Example 2-1 above, the synthesized single-element metal nanoparticles were analyzed using a Fourier transform infrared spectrophotometer (FT-IR) (Nicolet™ iS™50, Thermo Scientific, USA) in a range from 400 to 4000 cm−1 at room temperature.
As a result, as can be seen in
3-1: Synthesis of Metal Alloy Nanoparticles Using Recombinant Microorganism
One hour after inducing expression of metallothionein and phytochelatin synthase in the recombinant microorganism produced in Example 1 above by IPTG, i) cobalt and iron (Co, CoCl2, 0.5 mM, Fe, Fe (NO)3.6H2O, 0.5 mM), ii) nickel and iron (Ni, NiCl2.6H2O, 0.5 mM, Fe, Fe(NO)3.6H2O, 0.5 mM), iii) zinc and manganese (Zn, ZnSO4.7H2O, 0.5 mM, MnCl24H2O), 0.5 mM or iv) zinc and iron (Zn, ZnSO4.7H2O, 0.5 mM, Fe, Fe(NO)3.6H2O, 0.5 mM) ions, were added to the LB liquid medium (Tryptone 10 g/L, yeast extract 5 g/L, NaCl 5 g/L′ pH 6.5) in which the recombinant E. coli strain has been cultured, after which the recombinant microorganism was additionally cultured for 12 hours. The culture was centrifuged at 3500 rpm and 4° C. for 15 minutes, after which the supernatant was discarded and the E. coli pellet was collected. The E. coli pellet was washed three times with PBS buffer, and then dried in a freeze-dryer under vacuum for one day or more. Synthesis of nanoparticles was examined using transmission electron microscopy (TEM).
As a result, as shown in
Meanwhile, as shown in
3-2: Confirmation of Surface Functional Groups of Synthesized Metal Alloy Nanoparticles
In order to confirm the surface functional groups of the metal alloy nanoparticles synthesized in Example 3-1 above, the synthesized metal alloy nanoparticles were analyzed using a Fourier transform infrared spectrophotometer (FT-IR) (Nicolet™ iS™50, Thermo Scientific, USA) in a range from 400 to 4000 cm−1 at room temperature.
As a result, as can be seen in
3-3: Analysis of Magnetic Properties of Synthesized Metal Alloy Nanoparticles
In order to examine the magnetic properties of the nanoparticles produced in Example 3-1, magnetic hysteresis (M-H) curves of the nanoparticles were measured using an MPMS3 (magnetic property measurement system) (SQUID-VSM) (Quantum Design, USA) with SQUID-VSM function. Measurement of the M-H curves was performed at a temperature of 300K and at a magnetic field intensity of −50K Oe to +50K Oe.
As a result, as shown in
4-1: Synthesis of Metal Alloy Nanoparticles Using Recombinant Microorganism
One hour after inducing expression of metallothionein and phytochelatin synthase in the recombinant microorganism produced in Example 1 above by IPTG, silver (Ag, AgNO3, 0.25, 1 mM) and tellurium (Te, NaTeO3, 0.25, 1 mM) ions were added to the LB liquid medium (Tryptone 10 g/L, yeast extract 5 g/L, NaCl 5 g/L, pH 6.5) in which the recombinant E. coli strain has been cultured, after which the recombinant microorganism was additionally cultured for 12 hours. The culture was centrifuged at 3500 rpm and 4° C. for 15 minutes, after which the supernatant was discarded and the E. coli pellet was collected. The E. coli pellet was washed three times with PBS buffer, and then dried in a freeze-dryer under vacuum for one day or more. Synthesis of nanoparticles was examined using transmission electron microscopy (TEM).
As a result, as shown in
4-2: Confirmation of Surface Functional Groups of Synthesized Silver Tellurite (Ag2TeO3) Nanoparticles
In order to confirm the surface functional groups of the silver tellurite (Ag2TeO3) nanoparticles synthesized in Example 4-1 above, the silver tellurite (Ag2TeO3) nanoparticles were analyzed using a Fourier transform infrared spectrophotometer (FT-IR) (Nicolet™ iS™50, Thermo Scientific, USA) in a range from 400 to 4000 cm−1 at room temperature.
As a result, as can be seen in
4-3: Analysis of Magnetic Properties of Synthesized Silver Tellurite (Ag2TeO3) Nanoparticles
In order to examine the magnetic properties of the silver tellurite (Ag2TeO3) nanoparticles produced in Example 4-1, magnetic hysteresis (M-H) curves of the nanoparticles were measured using an MPMS3 (magnetic property measurement system) (SQUID-VSM) (Quantum Design, USA) with SQUID-VSM function. Measurement of the M-H curves was performed at a temperature of 300K and a magnetic field intensity of −70K Oe to +70K Oe.
As a result, as can be seen in
4-4: Analysis of Electrochemical Properties of Synthesized Silver Tellurite (Ag2TeO3) Nanoparticles
To analyze the electrochemical properties of the silver tellurite (Ag2TeO3) nanoparticles produced in Example 4-1, an electrochemical analyzer (Potentiostat; Princeton applied research, VSP) was used. The electrochemical properties of the nanoparticles were measured by cyclic voltammetry (CV) in 1M NaOH aqueous solution. To this end, the produced silver tellurite (Ag2TeO3) nanoparticles were dried on a glassy carbon electrode to form a working electrode. Furthermore, a platinum (Pt) wire was used as a counter electrode, and silver/silver chloride (Ag/AgCl) was used as a reference electrode. Measurement by cyclic voltammetry (CV) was performed at a voltage ranging from −0.4 to +0.5 V and at different scan rates of 10, 20, 30, 50 and 100 mV/s.
As a result, as shown in
5-1: Synthesis of Metal Sulfide Nanoparticles Using Recombinant Microorganism
One hour after inducing expression of metallothionein and phytochelatin synthase in the recombinant microorganism produced in Example 1 above by IPTG, sulfur (S, Na2S, 1 mM) together with silver (Ag, AgNO3, 1 mM), indium (In, InCl24H2O, 1 mM)), manganese (Mn, MnSO45H2O, 1 mM) or tin (Sn, SnCl24H2O, 1 mM) were added to the LB liquid medium (Tryptone 10 g/L, yeast extract 5 g/L, NaCl 5 g/L, pH 6.5) in which the recombinant E. coli strain has been cultured, after which the recombinant microorganism was additionally cultured for 12 hours.
The culture was centrifuged at 3500 rpm and 4° C. for 15 minutes, after which the supernatant was discarded and the E. coli pellet was obtained. The E. coli pellet was washed three times with PBS buffer, and then dried in a freeze-dryer under vacuum for one day or more. Synthesis of nanoparticles was examined using transmission electron microscopy (TEM) (Tecnai F20, Philips, Netherlands) and energy-dispersive X-ray spectroscopy (EDX) HD/MAX-2,500, Rigaku, Japan) with CuKa radiation (λ=1.5406 Å)).
As a result, as shown in
5-2: Confirmation of Surface Functional Groups of Synthesized Metal Sulfide Nanoparticles
In order to confirm the surface functional groups of the metal sulfide nanoparticles synthesized in Example 5-1 above, the synthesized metal sulfide nanoparticles were analyzed using a Fourier transform infrared spectrophotometer (FT-IR) (Nicolet™ iS™50, Thermo Scientific, USA) in a range from 400 to 4000 cm−1 at room temperature.
As a result, as can be seen in
In the case of some metals, it was shown that metal nanoparticles were not synthesized even when metal ions were added to the medium in which the recombinant microorganism produced in Example 1 had been cultured. In other words, one hour after inducing expression of metallothionein and phytochelatin synthase in the recombinant microorganism produced in Example 1 above by IPTG, cobalt (Co, CoCl26H2O, 0.5 mM), nickel (Ni, NiCl26H2O, 0.5 mM), zinc (Zn, ZnSO47H2O, 0.5 mM) or cadmium (Cd, CdCl2, 0.5 mM) ions, were added to the LB liquid medium (Tryptone 10 g/L, yeast extract 5 g/L, NaCl 5 g/L′ pH 6.5) in which the recombinant E. coli strain has been cultured, after which the recombinant microorganism was additionally cultured for 12 hours. The culture was centrifuged at 3500 rpm and 4° C. for 15 minutes, after which the supernatant was discarded and the E. coli pellet was collected. The E. coli pellet was washed three times with PBS buffer, and then dried in a freeze-dryer under vacuum for one day or more. In this procedure, whether nanoparticles would be synthesized was examined by transmission electron microscopy (TEM) (Tecnai F20, Philips, Netherlands), but the shape of synthesized nanoparticles could not be observed, and only the appearance of the recombinant E. coli was observed (
Accordingly, assuming that the ability of enzyme to bind to metal may vary depending on a change in pH, the present inventors changed pH to induce the synthesis of metal nanoparticles. In other words, one hour after inducing expression of metallothionein and phytochelatin synthase in the recombinant microorganism produced in Example 1 above by IPTG, the initial pH of the LB liquid medium (Tryptone 10 g/L, yeast extract 5 g/L, NaCl 5 g/L′ pH 6.5) in which the recombinant E. coli strain has been cultured is increased to 7.5, and then cobalt (Co, CoCl26H2O, 0.5 mM), nickel (Ni, NiCl26H2O, 0.5 mM), zinc (Zn, ZnSO47H2O, 0.5 mM) or cadmium (Cd, CdCl2, 0.5 mM) ions were added to the LB liquid medium, after which the recombinant microorganism was additionally cultured for 12 hours. After 12 hours of the culture, the pH of the medium was increased to 8 to 8.5. Thereafter, the culture was centrifuged at 3500 rpm and 4° C. for 15 minutes, after which the supernatant was discarded and the E. coli pellet was collected. The E. coli pellet was washed three times with PBS buffer, and then dried in a freeze-dryer under vacuum for one day or more. In this procedure, whether metal nanoparticles would be synthesized was examined by transmission electron microscopy (TEM) (Tecnai F20, Philips, Netherlands), and as a result, it could be seen that metal nanoparticles were synthesized, and particularly, the synthesized metal nanoparticles were crystalline (
One hour after inducing expression of metallothionein and phytochelatin synthase in the recombinant microorganism produced in Example 1 above by IPTG, lanthanum (La, La(NO3)36H2O, 0.5 mM), cerium (Ce, Ce(NO3)36H2O, 0.5 mM), praseodymium (Pr, Pr(NO3)36H2O, 0.5 mM), neodymium (Nd, Nd(NO3)36H2O, 0.5 mM), samarium (Sm, Sm(NO3)36H2O, 0.5 mM), europium (Eu, Eu(NO3)36H2O, 0.5 mM) or gadolinium (Gd, Gd(NO3)36H2O, 0.5 mM) ions were added to the LB liquid medium (Tryptone 10 g/L, yeast extract 5 g/L, NaCl 5 g/L, pH 6.5) in which the recombinant E. coli strain has been cultured, after which the recombinant microorganism was additionally cultured for 12 hours. The culture was centrifuged at 3500 rpm and 4° C. for 15 minutes, after which the supernatant was discarded and the E. coli pellet was collected. The E. coli pellet was washed three times with PBS buffer, and then dried in a freeze-dryer under vacuum for one day or more. In this procedure, whether nanoparticles would be synthesized was examined by transmission electron microscopy (TEM) (Tecnai F20, Philips, Netherlands), but it was shown that synthesized nanoparticles were all amorphous, and crystalline nanoparticles were not synthesized (
Accordingly, based on the fact that adjustment of the initial pH of the medium in the metal nanoparticle synthesis step induced the synthesis of crystalline metal nanoparticles which have not been synthesized, the present inventors adjusted the initial pH of the medium in the metal nanoparticle synthesis step. In other words, one hour after inducing expression of metallothionein and phytochelatin synthase in the recombinant microorganism produced in Example 1 above by IPTG, the initial pH of the LB liquid medium (Tryptone 10 g/L, yeast extract 5 g/L, NaCl 5 g/L′ pH 6.5) in which the recombinant E. coli strain has been cultured is increased to 7.5, and then lanthanum (La, La(NO3)36H2O, 0.5 mM), cerium (Ce, Ce(NO3)36H2O, 0.5 mM), praseodymium (Pr, Pr(NO3)36H2O, 0.5 mM), neodymium (Nd, Nd(NO3)36H2O, 0.5 mM), samarium (Sm, Sm(NO3)36H2O, 0.5 mM), europium (Eu, Eu(NO3)36H2O, 0.5 mM), or gadolinium (Gd, Gd(NO3)36H2O, 0.5 mM) ions were added to the LB liquid medium, after which the recombinant microorganism was additionally cultured for 12 hours. After 12 hours of the culture, the pH of the medium was increased to 8 to 8.5. Thereafter, the culture was centrifuged at 3500 rpm and 4° C. for 15 minutes, after which the supernatant was discarded and the E. coli pellet was collected. The E. coli pellet was washed three times with PBS buffer, and then dried in a freeze-dryer under vacuum for one day or more. In this procedure, whether nanoparticles would be synthesized was examined by transmission electron microscopy (TEM) (Tecnai F20, Philips, Netherlands), and as a result, it could be seen that amorphous metal nanoparticles were synthesized before pH adjustment, but crystalline metal nanoparticles were synthesized as a result of pH adjustment (
According to the present invention, a method for synthesizing metal nanoparticles is provided which have been difficult to synthesize by conventional biological methods. The present invention makes it possible to synthesize metal nanoparticles in an environmentally friendly and cost-effective manner, and also makes it possible to synthesize metal sulfide nanoparticles. In addition, even metal nanoparticles which could have been produced by conventional chemical or biological methods are produced in a significantly increased yield by use of the method of the present invention. In addition, the magnetic properties, the electrochemical properties, and the catalytic activity of the synthesized nanoparticles are identified so that the present invention can be applied to various industrial field applications including photoelectronics, batteries, contrast agents, semiconductor device, electronic devices, biosensors, catalysts, medicines, cosmetics, energy storage applications.
Although the present invention has been described in detail with reference to the specific features, it will be apparent to those skilled in the art that this description is only for a preferred embodiment and does not limit the scope of the present invention. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.
Number | Date | Country | Kind |
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10-2016-0002710 | Jan 2016 | KR | national |
10-2016-0182245 | Dec 2016 | KR | national |
Filing Document | Filing Date | Country | Kind |
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PCT/KR2016/015498 | 12/29/2016 | WO | 00 |