CN109694887A - Recombinant yeast and its purposes for manufacturing biomass alcohol - Google Patents
Recombinant yeast and its purposes for manufacturing biomass alcohol Download PDFInfo
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- CN109694887A CN109694887A CN201710998379.3A CN201710998379A CN109694887A CN 109694887 A CN109694887 A CN 109694887A CN 201710998379 A CN201710998379 A CN 201710998379A CN 109694887 A CN109694887 A CN 109694887A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
- C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
- C12P7/10—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate substrate containing cellulosic material
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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Abstract
The present invention provides the purposes that a kind of recombinant yeast is used to manufacture biomass alcohol, it includes: make yeast expressed artificial phytochelatin；Saccharomycete is set to contact biomass material, wherein biomass material includes carbohydrate and heavy metal ion；The artificial plant is set to chelate element in conjunction with the heavy metal ion；And saccharomycete is made to convert ethyl alcohol for the carbohydrate.
Description
Technical field
The present invention relates to the purposes of a Yeasts, and in particular to using recombinant yeast for manufacturing biomass alcohol
Purposes.
Background technique
In the industrial development of biomass alcohol, the main fermentation that biomass is carried out using saccharomycete, makes biomass at present
In the conversion of sugared part, therefore the ability of the growth situation of saccharomycete, production alcohol all significantly affects the production capacity of biomass alcohol.Generally
For, in the conversion reactor of biomass alcohol, the good growing environment of saccharomycete can be provided as far as possible, however, if making
Biomass material is to educate ground again from biology, then may contain a large amount of heavy metal ion in biomass material, lead to ferment
The growth of female bacterium and the conversion of alcohol are suppressed.
Existing document is attempted to imitate utilizes Escherichia coli (Escherichia coli, E.coli) big scale in the prior art
Up to natural chelating element using the concept as heavy metal absorbent, further turn plantation technology using gene and change to pass through saccharomycete
(yeast) plain (phytochelatin) is chelated to express the natural plants, and then heavy metal ion is made to influence saccharomycete
It is chelated before function, avoids the running for influencing processing procedure.However, by way of turning to grow expression natural plants chelating plain gene,
There are still the stereochemical structure complexity because of natural plants chelating element, and have steadily to have provided in successful translation, folding and modification
The problem of the natural plants chelating element of function.
Summary of the invention
Based on above-mentioned technical problem, the present invention provides the purposes that a kind of recombinant yeast is used to manufacture biomass alcohol, packet
It is plain containing the yeast expressed artificial plant with the structure as shown in chemical formula (1) is chelated:
Chemical formula (1)
Saccharomycete is set to contact biomass material, wherein biomass material includes carbohydrate and heavy metal ion, is made one
Work phytochelatin is in conjunction with heavy metal ion, and saccharomycete is made to convert ethyl alcohol for carbohydrate.
Preferably, saccharomycete surface expression or secreting, expressing artificial plant chelating element.
Preferably, heavy metal ion include copper ion, lead ion, nickel ion, cadmium ion, cobalt ions, zinc ion, mercury from
Son, silver ion or any combination thereof.
Preferably, saccharomycete is saccharomyces cerevisiae (Saccharomyces cerevisiae).
Preferably, biomass material educates ground again from biology.
Preferably, carbohydrate is selected from cellulose, hemicellulose and lignin.
Preferably, recombinant yeast is further used for joint bioconversion processing procedure.
In addition, the present invention provides a kind of recombinant yeast of purposes for above-mentioned manufacture biomass alcohol, it includes: in
The artificial plant with the structure as shown in chemical formula (1) of the recombinant yeast surface expression or secreting, expressing chelates element:
Chemical formula (1)
The present invention has an advantage that
1. the saccharomycete selected is eucaryote, compared to the prokaryotes of simple structure, it can more completely express and turn to grow
Extrinsic protein.
2. the present invention selects the mode of surface expression (surface display) or secreting, expressing compared to intracellular expression
It carries out turning to grow.The mode of surface expression is selected to have an advantage that (1) can effectively completely cut off comprising heavy metals such as cadmium, nickel, copper and lead
Ion chelating does not enter in saccharomycete body in outside yeast cell, therefore can avoid influencing the metabolism and growth situation of saccharomycete；
(2) chelating can be recycled in the heavy metal outside yeast cell, increase using recombinant yeast of the invention in manufacture biomass wine
The benefit of the purposes of essence more promotes the valence using the supplementary measures between the ecological restoration of the invention to be developed and biomass energy
Value.
3. the effect of artificial plant chelating plain (Synthetic phytochelatin, EC) is chelated to natural plants is similar,
However surface expression artificial plant chelating element is relatively simple in the processing procedure of saccharomycete, and has steady sources, easily industrialized
Mass production and the uniform advantage of quality, be more conducively used for the conversion processing procedure of biomass alcohol.In addition, also this can be recombinated
Saccharomycete, which is used as in joint bioconversion processing procedure (Consolidated Bio-Processing, CBP) at present, overcomes crucial problem
Technological means, the procedure for producing of further enhanced biological matter alcohol, and developing between a kind of ecological restoration and biomass energy
Effective supplementary measures.
In conclusion the recombinant yeast of expression artificial plant chelating element provided by the invention, with unexpressed saccharomycete
It compares, the inhibition situation of Heavy Metal Irons On Saccharomycescerevisiae bacterium growth can be reduced.Therefore, the present invention, which also provides, utilizes the recombination ferment
Female bacterium makes the biomass material comprising heavy metal ion be used to manufacture the purposes of biomass alcohol.
Detailed description of the invention
Fig. 1 is the flow chart of one embodiment of the invention.
Fig. 2 is the structural schematic diagram of the recombinant yeast of one embodiment of the invention.
Fig. 3 is the fluorescent microscopy images of one embodiment of the invention.
Fig. 4 is the adsorption analysis figure of one embodiment of the invention.
Fig. 5 is the cell growth curve figure of one embodiment of the invention.
Fig. 6 A and Fig. 6 B is respectively the biomass alcohol output and yield analysis chart of one embodiment of the invention.
Fig. 7 is the biomass alcohol output analysis chart of another embodiment of the present invention.
Specific embodiment
For make after above-mentioned purpose, technical characteristic and actual implementation it is incremental become apparent understandable, will be below
Corresponding relevant schema is assisted preferably to implement example to be described in detail.
In one embodiment of this invention, manufacturing process is as shown in Figure 1.
In step S10, a kind of recombinant yeast is established, because of the cell wall that saccharomycete has thickness enough and stable,
The protein of surface expression thereon can be made to maintain to stablize, and modification is good after its translation, the albumen for being suitble to performance complicated
Matter.And the method for yeast expressed foreign gene includes intracellular expression, secreting, expressing and surface expression etc..The mode of intracellular expression
It is to be accumulated on after expressing exogenous protein in the cytoplasm of saccharomycete；The mode of secreting, expressing is by exogenous protein point
It secretes in extracellular；And the mode of surface expression is then different from the above two, and the side of exogenous protein is expressed for a kind of immobilization
Method.The mode of surface expression be by the gene of the exogenous protein in conjunction with the gene of specific anchoring molecule after, turn grow into
Host cell translates exogenous protein together with anchoring molecule, and its secreting, expressing is made external source after outside cytoplasm
Property protein is anchored to host cell surface with anchoring molecule together.
In the present embodiment, select the mode of surface expression to establish recombinant yeast.Specifically, saccharomycete can be to appoint
What is usually used in the saccharomycete of alcoholic fermentation, such as: Schizosaccharomyces belongs to, Saccharomycodes belongs to,
Hanseniaspora belongs to saccharomycete etc., may preferably be brewer's yeast, is authenticated to be GRAS (generally
Recognized as safe) biology and safe biologic is confirmed as by U.S. FDA；And anchoring molecule then can be selected from it is general common
Comprising a agglutinin (such as: Aga1, Aga2), α agglutinin (such as: Ag α 1) with flocculate plain (FlolP) etc. anchoring molecule in,
When any artificial plant corresponding to recombinant yeast of the invention chelates element, affinity anchoring molecule.In addition, recombination
Saccharomycete may also respond with demand while it being made to express the catabolic enzyme of pentose, make it have the capacity of decomposition of pentose.
In addition, be usually used in chelated mineral metal-chelating protein include metallothionein (Metallothioneines,
MTs) with phytochelatin etc..Wherein, the ability of phytochelatin chelating heavy metal is better than metallothionein.However utilize yeast
When bacterium expresses phytochelatin, because phytochelatin is that ferment synthesis type wins peptide, it is caused to be not easy expressed intact in saccharomycete
Above and it is not easy to carry out one-step synthesis in solution.Therefore, this case is selected after saccharomycete is translated, similar to phytochelatin structure
Artificial plant chelate element.By in following chemical formula (1) and chemical formula (2) it is found that the change of the artificial plant chelating element after translation
It learns structure and contains alpha key, and the phytochelatin after translating then is gamma key, the difference in this structure neither will cause weight
The influence of metal adsorption ability, and the difficulty of processing procedure can be greatly reduced.
Chemical formula (1)
Chemical formula (2)
In step S20, the biomass material containing carbohydrate and heavy metal ion is added.Specifically, biological
Matter raw material is with may be from the ecological restoration by heavy metal pollution.Heavy metal may include copper ion, lead ion, nickel ion, cadmium ion,
Cobalt ions, zinc ion, mercury ion, silver ion or any combination thereof.And the type of biomass material can have cellulose to be any
Raw material.Because biology when educating again used plant different selections can be done according to the type of heavy metal pollution, however no matter select
Which kind of plant is selected, as long as having cellulose, can be used as biomass material.And biomass material also can be for by heavy metal pollution
Frequently seen plants, such as: barley, wheat, oat, rice, beet, sugar grass, the grain raw materials such as cassava and sweet Potato；Or
Non- grain raw material, such as: straw, rice straw, cornstalk etc.；Or agricultural, city and building castoff, such as food waste, newspaper, wood
Bits, culled wood etc.；Or the quick cellulosic crop of growth, such as Chinese silvergrass, Chinese pennisetum, switchgrass；Or the raw material for being easy to acquire,
Such as seaweed.
In step S31, recombinant yeast contacts with biomass material and manufactures biomass alcohol, meanwhile, step S32 will
Heavy metal ion is chelated in outside yeast cell.Saccharomycete manufactures the process of biomass alcohol, to have in technical field
The prior art of those of ordinary skill, therefore do not repeat herein.
In one embodiment of this invention, the structural schematic diagram of recombinant yeast is as shown in Figure 2.Wherein, saccharomycete is selected
The saccharomyces cerevisiae that Saccharomyces belongs to, anchoring molecule then select Ag α 1, that is, a kind of cell wall integral protein matter (cell
wall-anchored protein).Artificial plant is chelated to the sequence and α agglutinin (Ag α of plain (EC20) (SEQ ID NO:2)
1) it after (SEQ ID NO:3) is combined, is inserted into the genome of saccharomyces cerevisiae, penetrates artificial phytochelatin and Ag α 1 together
After secreting signal (MF α 1) (SEQ ID NO:1) secreting, expressing, it is anchored on saccharomyces cerevisiae cell wall as surface table
It reaches.And 1 gene of Ag α in saccharomyces cerevisiae genome is still kept completely, in order to avoid the structure of damage beer yeast cells wall and thin
The signal exchange of intercellular, therefore can avoid influencing the ability that saccharomycete is used to manufacture biomass alcohol.In addition, heavy metal ion with
The chelating of artificial plant chelating element is in progress outside yeast cell, therefore the growth of saccharomycete is not by the shadow of heavy metal depression effect
It rings.
In one embodiment of this invention, it by α agglutinin (Ag α 1) and is secreted first with polymerase chain reaction (PCR)
Signal (MF α 1) is expanded from saccharomyces cerevisiae.Recycling sticks together (annealing) and synthesizes the artificial phytochelatin of coding
(EC20) gene.This three genes are rejoined to the expression plastid of upper saccharomycete finally by restriction enzyme and ligase.Later,
First turn shape and turn shape after great expression plastid again into Escherichia coli into saccharomycete, artificial plant is chelated into plain (EC20) expression and is existed
On the surface of saccharomycete, the huge sum of money that can adsorb made belongs to surface.
Hereinafter, carrying out in above-described embodiment, the recombinant yeast of surface expression artificial plant chelating element manufactures biomass wine
The analysis of the purposes of essence, the purposes of biomass alcohol is manufactured to verify recombinant yeast of the invention with it, reaches effective production
The purpose of biomass alcohol.
Non- recombinant yeast of the invention is the brewer's yeast of the wild type (wild type) without any processing.Such as Fig. 3
It is shown, the recombinant yeast of surface expression artificial plant chelating element is marked with 488 fluorescent dye of Alexa Fluor.
Fig. 3 (A) and Fig. 3 (B) are respectively recombinant yeast and photo of the non-recombinant yeast under the bright visual field of microscope, display recombination ferment
The kenel of female bacterium and non-recombinant yeast.Fig. 3 (C) and Fig. 3 (A), which are shown, excites 488 fluorescence of Alexa Fluor under same field of view
The result of the wavelength observation of dyestuff, hence it is evident that observe the fluorescence being excited, represent artificial phytochelatin and be expressed in recombination really
The surface of saccharomycete.And Fig. 3 (D) and Fig. 3 (B) show the wavelength that 488 fluorescent dye of Alexa Fluor is excited under same field of view
Observation as a result, and have not seen any fluorescent signal, representative has no expression artificial plant chelating element on saccharomycete.
As shown in Fig. 4 and table 1, the analysis that above-described embodiment carries out adsorption capacity is connected.Select heavy metal cadmium (Cd) conduct
The pollution sources of simulation carry out recombinant yeast and non-recombinant yeast using inductively coupled plasma emission spectrometer (ICP-AES)
The analysis of adsorption capacity is carried out, to inquire into and measure the mode of absorption cadmium metal, and respectively with Langmuir equation (qm=
KbCe/(1+KCe), wherein qm is adsorbance, CeFor equilibrium concentration, b is maximal absorptive capacity, and K is that the adsorption equilibrium of adsorbate is normal
Number) and Freundlich equation (qm=KC1/n, wherein K and n are constant) and carry out the fittings (fitting) of data.Known to
The effect that Langmuir equation is fitted be it is preferred, and the adsorption capacity of recombinant yeast be non-recombinant yeast 2 again with
On.
Table 1
As shown in figure 5, connecting the analysis that above-described embodiment carries out Cell growth ability.It is found that recombinating ferment without in cadmium environment
Female bacterium and the growing state difference of non-recombinant yeast are little.However, in there are in the environment of 50 μM of cadmiums, the life of recombinant yeast
Long situation is substantially better than the growth situation of non-recombinant yeast.Heavy metal can be reduced to Yeast Growth by representing recombinant yeast
Inhibit.
As shown in figs. 6 a and 6b, it connects above-described embodiment and carries out biomass alcohol output and yield analysis.For educating again
The trees in soil, and using gas chromatograph (GC) to inquire into influence of the heavy metal cadmium for the efficiency of saccharomycete production alcohol.
As shown in Figure 6A, in no cadmium environment, the volume variance of recombinant yeast and non-recombinant yeast manufacture alcohol is little, as cadmium is dense
Degree is promoted, and the ability that recombinant yeast manufactures alcohol is more obvious better than the situation of the ability of non-recombinant yeast manufacture alcohol.
Yeast Growth can not be inhibited by heavy metal by representing recombinant yeast, and effectively manufacture biomass alcohol.And Fig. 6 B is
The yield analysis chart that concentration and time relationship according to Fig. 6 A calculate, display recombinant yeast and do not recombinate ferment without in cadmium environment
The yield difference of female bacterium manufacture alcohol is little, and as cadmium concentration is promoted, the ability of recombinant yeast manufacture alcohol is better than not recombinating
The situation that saccharomycete manufactures the ability of alcohol is more obvious.Representing recombinant yeast can not suppression by heavy metal to Yeast Growth
System, and effectively manufacture biomass alcohol.
As shown in fig. 7, connecting above-described embodiment, carrying out different types of heavy metal for recombinant yeast and not recombinating ferment
The analysis of the inhibition situation of female bacterium manufacture biomass alcohol.It is found that recombinant yeast no matter be in containing 150 μM of copper, 150 μM of nickel or
The environment of 150 μM of lead manufactures the yield of biomass alcohol still better than non-recombinant yeast, and representing recombinant yeast can not be by weight
Inhibition of the metal to Yeast Growth, and effectively manufacture biomass alcohol.
In summary, it was demonstrated that a kind of recombinant yeast of the invention is used to manufacture the purposes of biomass alcohol, can effectively drop
Inhibition of the low heavy metal for Yeast Growth, and improve there are when heavy metal environment, saccharomycete manufactures the tired of biomass alcohol
Border, is substantially improved that there are biomass alcohol output when heavy metal ion and yields, and develops a kind of ecological restoration and biomass
Effective supplementary measures between the energy.
Although the present invention specifically describes a kind of recombinant yeast of the invention for manufacturing biomass with above-described embodiment
The purposes of alcohol, however having that general technical staff of the technical field of the invention should be understood that can be without prejudice to skill of the invention
Under art principle and spirit, makes an amendment and change to embodiment.Therefore the scope of the present invention should be as claimed in claim.
[sequence table]
<110>Zhu Yimin
Changchun Artificial Resin Factory Co. Ltd.
Changchun Petrochemical Co., Ltd.
<120>recombinant yeast and its purposes for manufacturing biomass alcohol
<160> 3
<210> 1
<211> 267
<212> DNA
<213>brewer's yeast (Saccharomyces cerevisiae)
<220>
<223>signal (MF α 1) (Secretion signal) is secreted
<400> 1
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ccagtcaaca ctacaacaga agatgaaacg gcacaaattc cggctgaagc tgtcatcggt 120
tactcagatt tagaagggga tttcgatgtt gctgttttgc cattttccaa cagcacaaat 180
aacgggttat tgtttataaa tactactatt gccagcattg ctgctaaaga agaaggggta 240
tctctcgaga aaagagaggc tgaagct 267
<210> 2
<211> 126
<212> DNA
<213>artificial sequence
<220>
<223>artificial plant chelates plain (EC20), has the function of chelated metal ions.
<400> 2
atggaatgtg aatgtgaatg tgaatgtgag tgtgagtgtg agtgtgagtg tgagtgtgaa 60
tgcgaatgcg aatgcgaatg cgagtgcgag tgcgagtgcg agtgcgagtg cgagtgcgag 120
tgcggt 126
<210> 3
<211> 963
<212> DNA
<213>brewer's yeast (Saccharomyces cerevisiae)
<220>
<223>α agglutinin (Ag α 1)
<400> 3
agcgccaaaa gctcttttat ctcaaccact actactgatt taacaagtat aaacactagt 60
gcgtattcca ctggatccat ttccacagta gaaacaggca atcgaactac atcagaagtg 120
atcagccatg tggtgactac cagcacaaaa ctgtctccaa ctgctactac cagcctgaca 180
attgcacaaa ccagtatcta ttctactgac tcaaatatca cagtaggaac agatattcac 240
accacatcag aagtgattag tgatgtggaa accattagca gagaaacagc ttcgaccgtt 300
gtagccgctc caacctcaac aactggatgg acaggcgcta tgaatactta catctcgcaa 360
tttacatcct cttctttcgc aacaatcaac agcacaccaa taatctcttc atcagcagta 420
tttgaaacct cagatgcttc aattgtcaat gtgcacactg aaaatatcac gaatactgct 480
gctgttccat ctgaagagcc cacttttgta aatgccacga gaaactcctt aaattccttc 540
tgcagcagca aacagccatc cagtccctca tcttatacgt cttccccact cgtatcgtcc 600
ctctccgtaa gcaaaacatt actaagcacc agttttacgc cttctgtgcc aacatctaat 660
acatatatca aaacgaaaaa tacgggttac tttgagcaca cggctttgac aacatcttca 720
gttggcctta attcttttag tgaaacagca gtctcatctc agggaacgaa aattgacacc 780
tttttagtgt catccttgat cgcatatcct tcttctgcat caggaagcca attgtccggt 840
atccaacaga atttcacatc aacttctctc atgatttcaa cctatgaagg taaagcgtct 900
atatttttct cagctgagct cggttcgatc atttttctgc ttttgtcgta cctgctattc 960
taa 963
Claims (9)
1. the purposes that a kind of recombinant yeast is used to manufacture biomass alcohol, which is characterized in that it includes:
Make the yeast expressed artificial plant chelating element with the structure as shown in chemical formula (1):
Chemical formula (1)
So that the saccharomycete is contacted biomass material, wherein the biomass material include carbohydrate and heavy metal from
Son；
The artificial plant is set to chelate element in conjunction with the heavy metal ion；And
The saccharomycete is set to convert ethyl alcohol for the carbohydrate.
2. purposes as described in claim 1, which is characterized in that the saccharomycete is artificial plant chela described in surface expression
Close element.
3. purposes as described in claim 1, which is characterized in that the saccharomycete is artificial plant chela described in secreting, expressing
Close element.
4. purposes as claimed any one in claims 1 to 3, which is characterized in that the heavy metal ion be selected from copper ion,
In lead ion, nickel ion, cadmium ion, cobalt ions, zinc ion, mercury ion, silver ion and any combination thereof.
5. purposes as claimed any one in claims 1 to 3, which is characterized in that the saccharomycete is saccharomyces cerevisiae.
6. purposes as claimed any one in claims 1 to 3, which is characterized in that the biomass material is from biology
Educate ground again.
7. purposes as claimed any one in claims 1 to 3, which is characterized in that the carbohydrate be selected from cellulose,
Hemicellulose and lignin.
8. purposes as claimed any one in claims 1 to 3, which is characterized in that the recombinant yeast is further used
In joint bioconversion processing procedure.
9. a kind of recombinant yeast, which is characterized in that it includes:
There is the artificial plant chela of the structure as shown in chemical formula (1) in the recombinant yeast surface expression or secreting, expressing
Close element:
Chemical formula (1)
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Family
ID=66226233
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009026680A1 (en) * | 2007-08-27 | 2009-03-05 | Ping I Lee | Supramacrqmolecular polymer complexes providing controlled nitric oxide release for healing wounds |
| CN101454454A (en) * | 2006-05-22 | 2009-06-10 | 韩国科学技术院 | Method for preparing metal nanoparticles using metal binding proteins |
| CN103626847A (en) * | 2013-11-15 | 2014-03-12 | 江南大学 | Wheat germ protein source zinc phytochelatin and preparation method thereof |
-
2017
- 2017-10-24 CN CN201710998379.3A patent/CN109694887A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101454454A (en) * | 2006-05-22 | 2009-06-10 | 韩国科学技术院 | Method for preparing metal nanoparticles using metal binding proteins |
| WO2009026680A1 (en) * | 2007-08-27 | 2009-03-05 | Ping I Lee | Supramacrqmolecular polymer complexes providing controlled nitric oxide release for healing wounds |
| CN103626847A (en) * | 2013-11-15 | 2014-03-12 | 江南大学 | Wheat germ protein source zinc phytochelatin and preparation method thereof |
Non-Patent Citations (3)
| Title |
|---|
| CHI-EN YANG，等: "Surface display of synthetic phytochelatins on Saccharomyces cerevisiae for enhanced ethanol production in heavy metal-contaminated substrates", 《BIORESOURCE TECHNOLOGY》 * |
| ZHAOHUI XU，等: "Heavy Metal Removal by Novel CBD-EC20 Sorbents Immobilized on Cellulose", 《BIOMACROMOLECULES》 * |
| 徐照丽,等: "重金属植物螯合肽（PC）的研究进展", 《农业环保保护》 * |
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