644 results about "Heterologous expression" patented technology

The invention belongs to the technical field of protein engineering, and discloses a Brazilian sweet protein mutant and application thereof. According to the invention, lysine is used for replacing aspartic acid at the 29th site of Brazilian sweet protein, and / or arginine is used for replacing glutamic acid at the 36th site, and / or alanine is used for replacing aspartic acid at the 50th site, amino acid mutation at the sites is carried out to construct a Brazilian sweet protein mutant, and the Brazilian sweet protein mutant is subjected to heterologous expression in pichia pastoris GS115, so that the Brazilian sweet protein mutant is obtained. And carrying out sweetness performance evaluation on the expressed Brazilian sweet protein mutant. The sweetness property of the double mutant Asp29Lys-Asp50Ala protein is improved by 240% compared with the sweetness property of the Brazilian sweet protein before mutation.

The invention provides a Chinese cabbage clubroot susceptibility related gene BrSWEET14a, a construction method and application, belongs to the technical field of plant genetic engineering, and particularly relates to a Chinese cabbage clubroot susceptibility related gene BrSWEET14a. The CDS sequence of the Chinese cabbage BrSWEET14a is as shown in SEQ ID No. 1 (sequence identifier number 1). The gene is transformed into Col-type arabidopsis thaliana through an agrobacterium flower immersion transformation method, a BrSWEET14a heterologous expression arabidopsis thaliana strain is obtained, and the result shows that T-DNA insertion mutant of the BrSWEET14a homologous gene Atsweet14 is more resistant to clubroot than that of the Col-type arabidopsis thaliana, and heterologous expression of Chinese cabbage BrSWEET14a leads to reduction of disease resistance of arabidopsis thaliana to plasmodiophora brassicae. Therefore, the Chinese cabbage BrSWEET14a is closely related to the clubroot disease, and the gene and the homologous gene of the gene in other cruciferous plants can be applied to breeding of the cruciferous plants and other horticultural plants and have a good application prospect.

The invention provides a Ustilaginoidea virens gene UvASE2 for regulating and controlling rice resistance and application of the Ustilaginoidea virens gene UvASE2, and relates to the technical field of plant genetic engineering. The nucleotide sequence of the Ustilaginoidea virens gene UvASE2 is as shown in SEQ ID NO: 1, and the amino acid sequence of the encoding protein of the Ustilaginoidea virens gene UvASE2 is as shown in SEQ ID NO: 2. By constructing a transgenic rice plant for heterologous expression of UvASE2, it is found that the transgenic rice can significantly improve the resistance to false smut and bacterial blight and significantly induce the outbreak of active oxygen. The heterologous expression UvASE2 gene of Ustilaginoidea virens has the function of positively regulating and controlling the disease resistance of rice, and the UvASE2 gene can be used for improving the disease resistance of rice and has important significance on creation of disease-resistant germplasm of rice.

The invention discloses a schizochytrium limacinum genetic engineering strain capable of remodeling an FAS (Fibroblast Activated Sequence) pathway as well as a construction method and application thereof. The schizochytrium limacinum gene engineering strain is obtained by heterologous expression of a C16 elongase gene E16, a C18 elongase gene E18, a delta 9 desaturase gene D9, a delta 5 desaturase gene D5, a delta 6 desaturase gene D6 and a delta 15 desaturase gene D15 in a schizochytrium limacinum wild type. The recombinant schizochytrium limacinum described herein is capable of producing EPA comprising 2.08% by total fatty acids while also producing DHA comprising at least more than 41% by total fatty acids. In addition, the invention discloses a method for producing unsaturated fatty acid through a double-path channel of PKS and FAS, which is beneficial to promotion and application of a technology for producing EPA and DHA by schizochytrium limacinum.

The invention provides an NMN efficient synthesis strain and a construction method and production application thereof, and belongs to the technical field of engineering bacteria. According to the invention, firstly, an NMN synthesis route taking NAM as a substrate is constructed in escherichia coli, and specifically, NadV and PrsD128A are introduced into the escherichia coli, so that the NMN synthesis flux is improved; pnuC is introduced to promote the external arrangement of the NMN; the method comprises the following steps: knocking out PncC, NadR, UshA, NudC and MazG; pfkA and TalA are knocked out to improve the carbon flux of the pentose phosphate pathway. GDH1 and GDH2 are heterologously expressed in engineering bacteria to establish a reducing power balance system, the reducing power balance system is used for converting NADPH excessively accumulated in an NMN synthesis route into NADH, and meanwhile, an esaR-esaI group response element is introduced to dynamically regulate and control the system. The engineering strain constructed by the method can accumulate 3.0 g / L of NMN after 24-hour shake flask fermentation.

This application relates to the field of biosynthesis technology, and specifically discloses a pyridone compound, its biosynthesis method and application. Based on synthetic biology-related technologies, this application modifies non-ribosomal peptide synthetase, heterologously expresses non-ribosomal peptide synthetase mutants in a variety of hosts, and for the first time discovers and obtains a pyridone compound, and determines its chemical formula as C5H4N2O3, named 5-amino-pyridine-2,3,6-trione or 5-hydroxy-3-imino-pyridine-2,6-dione. This novel pyridone compound can be used as a raw material or intermediate in the development and preparation of novel antibiotics, drugs and dyes.

The invention discloses a 3 '-phosphoadenosine-5'-phosphoryl sulfate synthetase mutant and an application of the 3 '-phosphoadenosine-5'-phosphoryl sulfate synthetase mutant. Compared with a wild enzyme amino acid sequence as shown in SEQ ID NO.1, the amino acid sequence of the 3 '-adenosine-5'-phosphoryl sulfate synthetase mutant has one or two mutations in the 101 site, the 207 site and the 560 site; preferably, phenylalanine at the 101 site is mutated into tryptophan; cysteine at the 207th site is mutated into glycine; phenylalanine at the 560th site is mutated into tryptophan. The invention also provides a synthesis method and application of the 3 '-adenosine-5'-phosphoryl sulfuric acid. The APS intermediate product inhibition effect of hPAPSS1 is relieved through a point mutation technology, the substrate affinity of hPAPSS1 to ATP is improved, and a mutant enzyme with a high PAPS conversion rate is obtained; and the bifunctional enzyme hPAPSS1 is synthesized by heterologous expression of PAPS in escherichia coli, so that the acquisition of the enzyme is simplified, and the synthesis efficiency of PAPS is improved.

The invention relates to an L-valine production strain as well as a construction method and application thereof, the strain is obtained by transformation through a metabolic synthesis path of L-valine, and the construction method comprises the following steps: by taking Cornebacterium glutamicum ATCC 13032 (C.glutamicum) as an original strain, carrying out editing transformation such as gene knockout, gene overexpression, gene heterologous expression, gene double-copy expression and the like on a genome of the original strain by utilizing a metabolic engineering means. The L-valine production strain disclosed by the invention can be used for synthesizing L-valine from the beginning by taking glucose as a carbon source, has the advantages of high production rate, short fermentation period, high strain stability and low cost, and lays a foundation for realizing large-scale production of L-valine. According to the fermentation tank fermentation method, the L-valine is produced in a mechanical stirring type fermentation tank by using the L-valine production strain Va1-09, 86.4 g / L of L-valine is produced by fermenting for 36 hours, and the method has a good industrial prospect.

An artificial synthesis method for malonyl-CoA and use thereof are provided. By means of heterologous expression of an aminotransferase and a malonyl-CoA reductase, an artificial synthesis pathway for synthesizing malonyl-CoA by using β-alanine (β-ala) as a precursor is constructed as follows: firstly, under catalysis of a transaminase, β-ala transfers amino groups to α-ketonic acid (such as pyruvic acid, oxaloacetic acid, or α-ketoglutaric acid, etc.), to form an intermediate product 3-oxopropanoate and a corresponding amino acid; the 3-oxopropanoate generates malonyl-CoA under the action of the malonyl-CoA reductase. This pathway addresses the defects of the natural malonyl-CoA synthesis pathway, such as low carbon utilization, consumption of energy substance ATP, release of greenhouse gas CO2, and strict regulation of pathway enzymes, a pyruvate dehydrogenase (PDH) and an acetyl-CoA carboxylase (ACC), thereby achieving high yielding of products using malonyl-CoA as a precursor, including flaviolin, octanoic acid, phloroglucinol, pentadecaheptaene, natamycin, and spinosad.

The invention relates to the technical field of biology, in particular to glycopeptide antibiotic and application thereof. According to the invention, a biosynthetic gene cluster for synthesizing a brand new glycopeptide antibiotic skeleton is excavated, and brand new glycopeptide antibiotics Varsomycin A, B and C with novel chemical structures are obtained through heterologous expression; the traditional Chinese medicine composition has a remarkable antibacterial effect on clinically drug-resistant strains. Important fermentation strains and precursor compound molecules are provided for research and development of new drugs of glycopeptide antibiotics, and important application prospects and economic values are achieved.

The invention discloses a recombinant strain for producing squalene as well as a construction method and application of the recombinant strain. The construction method comprises the following steps: heterologously expressing cDNA (complementary deoxyribonucleic acid) of each gene in a squalene synthesis route from filamentous fungi in yeast; the squalene synthetic pathway genes comprise a mevalonic acid-5-phosphate kinase gene, a squalene synthase gene, an acetoacetyl coenzyme A thiolase gene, a mevalonic acid kinase gene, a hydroxymethyl glutaryl coenzyme a synthetase gene, a mevalonic acid pyrophosphate decarboxylase gene, a farnesyl pyrophosphate synthetase gene and isopentenyl pyrophosphate. A dimethallylpyrophosphate isomerase gene and a truncated hydroxymethyl glutaryl coenzyme a reductase gene; the culture medium is characterized in that the filamentous fungi are Saroocladium oryzae or Stibella sp. 1056, and the culture medium is a culture medium. The excavated squalene synthesis route can complete efficient de novo synthesis of squalene, and the squalene yield is increased.

The invention discloses a fusion protein which comprises a first structural domain with an EgtD catalytic function, a second structural domain with an Egt1 catalytic function and a third structural domain with an EgtE catalytic function, and the amino acid sequence of the first structural domain with the EgtD catalytic function is shown as SEQ ID NO.1; the amino acid sequence of the second structural domain with the Egt1 catalytic function is as shown in SEQ ID NO. 2; and the amino acid sequence of the third structural domain with the EgtE catalytic function is as shown in SEQ ID NO. 3. The fusion protein is subjected to heterologous expression in engineering bacteria, so that the yield of ergothioneine is remarkably increased.

The invention discloses a C1, 25-bit dihydroxylase mutant of vitamin D3 as well as a genetically engineered bacterium and application thereof, and belongs to the technical field of enzyme engineering. The preparation method comprises the following steps: mutating 81-site glutamic acid of vitamin D3 dihydroxylase of bacillus megatherium H-1 into alanine to obtain a vitamin D3 dihydroxylase mutant; pMA5 is used as an expression plasmid to realize heterologous expression of the vitamin D3 dihydroxylase mutant in the bacillus subtilis; when the engineering bacteria disclosed by the invention are applied to calcitriol synthesis, the mutant has the effects of increasing the concentration accumulation amount of a product and shortening the reaction time consumption, and the production requirements of shortening the time cost in industrial production are met.

The invention discloses a pichia pastoris engineering bacterium for synthesizing bakuchiol from de novo by converting methanol as well as construction and application of the pichia pastoris engineering bacterium. A heterologous p-coumaric acid synthesis pathway is introduced to neutral sites of different chromosomes of pichia pastoris, a high-yield p-coumaric acid strain is obtained through gene knockout, overexpression or heterologous expression of tyrosine and phenylalanine synthesis pathway key genes of a shikimic acid pathway, a heterologous bakuchiol biosynthesis pathway is introduced on the basis, and a high-yield p-coumaric acid strain is obtained. After overexpression of bakuchiol synthetase, endogenous overexpression or heterologous expression of MVA pathway key genes and exogenous acetyl coenzyme A supply pathway genes, methanol concentration optimization and bacterial strain His4 gene supplementation, the yield of bakuchiol is effectively increased to 91.2 mg / L and is increased by 59.8 times compared with that of an initial bacterial strain, and the yield of bakuchiol in a 15L fermentation tank reaches 692.8 mg / L. The method has the characteristics of high conversion efficiency, low production cost, convenience in preparation, wide industrial application prospect and the like.

The present invention discloses an antibacterial peptide cecropin A mutant, a recombinant expression vector, a Bacillus subtilis engineering bacterium and applications thereof. By modifying the antibacterial peptide cecropin A, the present invention screened mutants with antibacterial activities superior to those of the original cecropin A, and through the construction of a recombinant expression vector, heterologous expression was carried out using the biosafe strain Bacillus subtilis. Compared with the original antibacterial peptide cecropin A, the antibacterial activities of the mutants of the present invention were increased by 60% and 80% respectively. The present invention uses Bacillus subtilis as a secretion expression system to express the antibacterial peptide cecropin A mutant, which is conducive to large-scale industrial production and cost savings.

The invention relates to recombinant trichoderma reesei as well as a preparation method and application thereof, and belongs to the technical field of gene engineering and enzyme engineering. The recombinant trichoderma reesei comprises a promoter and a xylanase gene, the promoter is derived from trichoderma reesei, and the xylanase gene is derived from penicillium oxalicum. Trichoderma reesei is transformed by using a xylanase gene expression plasmid, heterologous expression of xylanase from penicillium oxalicum is realized, and feruloyl oligosaccharide is prepared by using the obtained xylanase and taking brewer's grains as a raw material, so that the feruloyl oligosaccharide has high yield.

A method for separating an exosome from a heterologous expression cell or a primary culture cell, and a use of an exosome in biological analysis, relating to the technical field of biomedicine. The method for separating the exosome from the heterologous expression cell or the primary culture cell of the present invention comprises: carrying out differential centrifugation on a solution to be extracted that contains an exosome, then taking out a first supernatant, carrying out high-speed centrifugation again, respectively adding iodixanol into an obtained second supernatant and a precipitate, and carrying out spontaneous density gradient centrifugation to obtain a separated purified exosome. According to the present invention, an iodixanol spontaneous density gradient centrifugation method is innovatively used to successfully carry out separation and purification to obtain the exosome having high purity and high yield, and the separated exosome is used as an experimental development universal platform for researching a membrane protein structure and functional characteristics in an original ecological cell membrane environment, and is used for biological analysis, drug screening, and disease diagnosis and treatment.

The invention relates to the technical field of gene engineering and recombinase catalysis engineering, in particular to a recombinant strain for expressing phenolic acid decarboxylase as well as a construction method and application of the recombinant strain. According to the invention, a recombinant strain for expressing the phenolic acid decarboxylase PADCs is constructed by heterologous expression of a coding gene PADCs of the phenolic acid decarboxylase. The enzyme liquid of the recombinant strain for expressing the phenolic acid decarboxylase has remarkable catalytic performance, p-coumaric acid can be used as a substrate, the recombinant strain is used for inducing expression, the purified enzyme liquid is used for catalyzing decarboxylation reaction of p-coumaric acid, and p-hydroxystyrene is synthesized; meanwhile, the phenolic acid decarboxylase PADCs has a wide substrate spectrum, the 4-vinylphenol compound can be used as a substrate, bicarbonate is used as a carboxyl source, the freeze-dried whole cell of the recombinant strain is utilized to catalyze the 4-vinylphenol compound to carry out regioselective beta-carboxylation reaction, a series of phenolic acids with biological activity are synthesized by an enzymic method, and meanwhile CO2 is immobilized; and a huge application potential is provided for the utilization of CO2.

The invention discloses recombinant silk fibroin peptide as well as a preparation method and application thereof, and belongs to the technical field of cosmetic raw material development. The amino acid sequence of the recombinant silk fibroin peptide SFHR6 disclosed by the invention is as shown in SEQ ID NO. 1. Soluble expression of the recombinant silk fibroin peptide is achieved through prokaryotic expression and eukaryotic expression, the high-purity soluble recombinant silk fibroin peptide SFHR6 is obtained through nickel column affinity chromatography, hydrophobic chromatography, cation chromatography and anion chromatography technologies, and the recombinant silk fibroin peptide SFHR6 is high in purity, high in tyrosinase inhibition rate and strong in whitening effect. The purification process provided by the invention is simple and convenient, has good repeatability, effectively solves the quality control problem of naturally extracted protein products, has the advantages of high efficiency, high-purity heterologous expression and low large-scale production cost, and has relatively high industrial application potential.

The invention discloses an application of transmembrane transporters NPF and SWEET in increasing the yield of gibberellin of Fusarium fujikuroi, and the application comprises the following steps: performing heterologous expression on an exogenous transmembrane transporter NPF or SWEET in wild type Fusarium fujikuroi in a manner of constructing an integrated expression plasmid, so as to increase the yield of gibberellin GA4 + 7. A new thought is provided for increasing the yield of gibberellin, the yield of GA4 + 7 is effectively increased through heterologous overexpression of the transmembrane transporter NPF or SWEET, gibberellin which is more efficient, more stable, safer and higher in economic value is obtained, and the gibberellin has a positive promotion effect on industrial production of gibberellin.

The present invention relates to means for recombinant manufacture. In particular, it relates to a recombinant bacterial cell comprising in its genome at least one essential gene the endogenous promoter of which has been replaced by a heterologous expression control sequence being inducible by an inducer molecule, such that the expression of said essential gene in the recombinant bacterial cell is dependent on the presence of said inducer molecule. Further contemplated are methods for generating the recombinant bacterial cell of the invention as well as a method for recombinant manufacture of a compound of interest. The present invention also provides for use the recombinant bacterial cell of the invention for recombinant manufacture of a compound of interest and a kit for manufacture of a compound of interest.

The invention provides a plasmid, an alpha-ketoglutaric acid engineering strain and a construction method and application of the alpha-ketoglutaric acid engineering strain, the strain is obtained by adopting a directional transformation method, transcriptional levels of katE, katG, sodB and grxA genes are up-regulated, a catR gene and a katA gene are heterologously expressed, a high-copy plasmid pETKG is carried, and the plasmid simultaneously expresses L-glutamate oxidase LGOX genes from Streptomyces sp and Chain A; the strain self-synthesized catalase eliminates H2O2 generated in the process of producing alpha-ketoglutaric acid through enzyme catalysis, engineering bacteria of L-glutamate oxidase LGOX expressed by pETKG plasmids are used for performing whole-cell catalysis on L-sodium glutamate to synthesize alpha-ketoglutaric acid, and the engineering strain is a high-level engineering strain capable of industrially and stably producing alpha-ketoglutaric acid.

The invention discloses production and application of an antibacterial protein targeting enterococcus pluvialis. Specifically, the antibacterial protein comprises an amino acid sequence as shown in any one of SEQ ID NO.15-28 or an active fragment and an analogue of the amino acid sequence. According to the present invention, the potential antibacterial protein is found from the genome of the bacteria or the bacteriophage, and the antibacterial protein can inhibit the enterococcus pluvialis in the targeting manner; the antibacterial protein with biological activity can be obtained in a heterologous expression mode and can remove the enterococcus pluvialis in a targeted mode, and then related diseases caused by the enterococcus pluvialis are relieved and treated. The antibacterial protein provided by the invention can perform targeted sterilization without damaging normal human symbiotic flora, and can avoid side effects caused by traditional antibiotic treatment.

The invention discloses application of a cellulose synthase-like gene GhCSLC4 in improvement of cotton yield traits, and belongs to the field of biotechnology application. According to the invention, a biotechnology is utilized to determine that the GhCSLC4 gene has a promotion effect on fiber cell elongation. Researches show that the silent cotton cellulose synthase gene GhCSLC4 inhibits fiber initiation and elongation, and the root length and the number of stem hairs of an arabidopsis plant with heterologous expression of GhCSLC4 are both higher than those of a wild control. The GhCSLC4 gene has important effects and application prospects in improvement of cotton fiber yield traits and cultivation of new high-yield cotton varieties.

The invention relates to pichia pastoris engineering bacteria and a preparation method thereof, and a method for producing cellooligosaccharide / disaccharide by using corn straws or wheat straws, and belongs to the technical field of genetic engineering. A cellulose incision enzyme gene and a cellulose excision enzyme gene are transferred into pichia pastoris engineering bacteria. The preparation method comprises the following steps: respectively inserting a cellulose incision enzyme gene and a cellulose excision enzyme gene into plasmids to construct expression vectors; transferring the expression vector into a competent cell of pichia pastoris GS115; and screening and identifying to obtain successfully transformed pichia pastoris engineering bacteria. According to the present invention, the pichia pastoris heterologous expression cellulose incision enzyme and cellulose excision enzyme are adopted to construct the pichia pastoris engineering strain capable of degrading the plant straw into the cellooligosaccharide / disaccharide, such that the reliable chassis strain is provided for the production of the starch from the cellulose;

The invention discloses a Pseudomonas putida high-yield rhamnolipid fermentation medium based on a step-by-step optimization strategy, an optimization method and application thereof, through the step-by-step strategy of component screening-single-factor optimization-multi-factor interactive optimization, a mixed carbon source and a mixed nitrogen source are taken as cores, and a Plackett-Burman test, a steepest climbing test and Box-Behnken response surface design are combined, so that the high-yield rhamnolipid fermentation medium for the pseudomonas putida high-yield rhamnolipid is obtained. Optimizing to obtain a special fermentation culture medium; two genetic engineering strains estA-2 and estA-3 are constructed by heterologous expression of key genes, and the supply and extracellular transport efficiency of a rhamnolipid synthesis precursor is enhanced; the recombinant strain is applied to the optimized culture medium, and efficient production of a shake flask and a 5L fermentation tank is achieved. The rhamnolipid yield of the estA-3 strain in a 5L fermentation tank reaches 35.59 + / -2.18 g / L, the conversion rate is 0.282 g / g, the productivity is 0.404 g / L / h, the rhamnolipid yield, the conversion rate and the productivity are obviously superior to those in the prior art, and a key technical support is provided for industrial production of rhamnolipid.

The application discloses a kind of fungal laccase mutant Lcc5-G and its expression strain and application.The application is based on the fungal laccase from Coprinopsis cinerea heterologous expression since, by site-directed mutagenesis, obtain mutant gene.After fungal induction expression containing mutant plasmid, obtain laccase mutant enzyme Lcc5-G with improved specific activity, stability and AFB1 removal efficiency.When ABTS is used as substrate, the specific activity of the mutant is improved by 3 times, and the stability of the mutant enzyme is improved to 5 times of the original enzyme at 45 DEG C and pH 7.The mutant has potential application value in oxidizing aflatoxin.

The invention belongs to the technical field of genetic engineering, and discloses a schizochytrium limacinum genetic engineering strain capable of reducing the proportion of DPA in grease and correspondingly increasing the content of DHA, a method and application of the schizochytrium limacinum genetic engineering strain. The strain is characterized in that an omega-3 desaturase gene derived from Platyneria dumeriii and / or Lepeophtheria salmonis is subjected to heterologous expression, so that the proportion of DPA (docosahexaenoic acid) in grease is reduced, and the content of DHA (docosahexaenoic acid) is correspondingly increased. According to the invention, by heterologous expression of Pd delta 19Des and Ls delta 19Des genes in schizochytrium limacinum, desaturation activity of two desaturases to a delta 19 position is verified, and conversion from omega-6 type DPA to omega-3 type DHA is effectively promoted.

The invention belongs to the technical field of gene engineering, and discloses recombinant yarrowia lipolytica as well as a construction method and application thereof. The recombinant yarrowia lipolytica PGA1 is obtained by carrying out heterologous expression on an arabidopsis thaliana inositol oxygenase gene and a pseudomonas syringae uronic acid dehydrogenase gene in the yarrowia lipolytica by utilizing a genetic engineering technical means. The recombinant yarrowia lipolytica PGA10 capable of efficiently synthesizing glucaric acid (GA) is finally obtained by further modifying six genes of related metabolic pathways of the recombinant yarrowia lipolytica PGA1. The GA yield of the PGA10 reaches 2233.24 mg / L and is 66.27 times that of PGA1, and the effect is remarkable. The invention not only constructs a GA synthesis method, but also defines the influence of related genes such as opi1 on GA synthesis, and provides reference for the research of heterologous synthesis of GA by microorganisms.

The invention discloses ethanol dehydrogenase, recombinant genetically engineered bacteria and application of the ethanol dehydrogenase in asymmetric reduction of prochiral carbonyl compounds, the ethanol dehydrogenase has good biological performance, heterologous expression in Escherichia coli engineering bacteria is achieved, intracellular coenzyme can be directly utilized when whole cells are adopted as a catalyst, and the catalyst has a good application prospect. Zero addition of the coenzyme is realized, the enzyme has the advantages of high activity, high stereoselectivity, wider substrate spectrum and the like, prochiral ketone compounds can be greenly and efficiently catalyzed to be converted into corresponding chiral alcohols, and the problems of low conversion rate, low yield, low e.e. Value, complex reaction process, more byproducts and the like in a traditional chemical method are solved. The ethanol dehydrogenase has good tolerance to isopropanol, and the isopropanol can be used as a hydrogen source to realize the circulation of coenzyme. And the byproduct acetone generated by the enzyme catalytic reaction and the isopropanol remaining in the reaction can be separated and recovered by a technical means, so that the production cost is greatly reduced.