53 results about "P450 Enzymes" patented technology

The invention relates to the technical field of agricultural biology, in particular to a fusion P450 enzyme mutant, a method for improving VD3 catalytic activity and a method for generating 25 (OH) VD3 through whole cells. The invention constructs a fusion P450 enzyme VK1-CYP116B46-L21 and a self-sufficiency fusion P450 enzyme mutant with improved activity, wherein the self-sufficiency fusion P450 enzyme mutant is used for catalyzing vitamin D3 to synthesize calcifediol; vD3 is taken as a substrate, glucose, a cosolvent and a wet thallus induced by recombinant escherichia coli for simultaneously expressing fusion P450 enzyme and glucose dehydrogenase are added, and the highest yield of 25 (OH) VD3 within 24 hours after catalytic reaction can reach 3.26 g / L; whole cells are directly used as a catalyst, enzyme purification is not needed, glucose dehydrogenase is introduced, and glucose is used as a substrate to regenerate a cofactor NAD (P) H, so that addition of expensive cofactors is avoided, the production cost is reduced, and industrial production is facilitated.

The application discloses a P450 enzyme mutant for degrading herbicides and application thereof, and the P450 enzyme mutant is named P450 BSβ -F46A, P450 BSβ -F46A-CPR; the amino acid sequences are shown in SEQ ID NO. 2 and 4 respectively. The application also discloses application of the P450 enzyme mutant in degrading pyridine oxycarboxylic acid herbicides or aryloxyphenoxypropionic acid herbicides and related genes in cultivating herbicide-resistant transgenic crops. Experiments prove that the highest conversion rate of the P450 enzyme mutant for degrading pyridine oxycarboxylic acid herbicides reaches 86%, and the total conversion number of catalyzing flumiclorac can reach 7443 at most, and the highest catalytic conversion rate of degrading aryloxyphenoxypropionic acid herbicides reaches 89%. The P450 enzyme mutant provided by the application has the advantages of high catalytic efficiency, high protein expression amount and low industrial cost, and the whole operation process for degrading herbicides is simple, the process is mature, the cost is low, and the application prospect is wide.

The present application relates to the field of agricultural biotechnology, in particular to fusion P450 enzyme mutants, methods for improving the catalytic activity of VD3, and methods for producing 25(OH)VD3 by whole cells, wherein a fusion P450 enzyme VK1-CYP116B46-L21 and a self-sufficient fusion P450 enzyme mutant with improved catalytic activity for synthesizing calcidiol from vitamin D3 are constructed; using VD3 as a substrate, adding glucose, a cosolvent, and a recombinant Escherichia coli expressing the fusion P450 enzyme and glucose dehydrogenase after induction, the maximum yield of 25(OH)VD3 can reach 3.26 g / L within 24 h after the catalytic reaction; directly using whole cells as catalysts without purifying enzymes, introducing glucose dehydrogenase to regenerate cofactor NAD(P)H using glucose as a substrate, avoiding the addition of expensive cofactors, reducing production costs, and being conducive to industrial production.

The application belongs to the technical field of bio-chemical industry, and particularly relates to a P450 enzyme mutant and application of an engineering bacterium thereof in synthesis of calcifediol, the mutant is one or more point positions in an amino acid sequence based on a wild type P450 enzyme Vdh being mutated; the amino acid sequence of the wild type P450 enzyme Vdh corresponds to GenBank accession number CP069288.1. Through systematic molecular dynamics simulation, conservation analysis and substrate binding pocket engineering on the Vdh enzyme, multiple key mutation sites are successfully screened and obtained. The constructed combined mutant (such as I114R / N173M / Q310R, Vdh-M3) shows extremely high hydroxylation activity, and the pure enzyme activity can reach 9.8 times of the wild type enzyme, greatly improving the conversion rate from vitamin D3 to calcifediol.

The present disclosure relates to relates to compounds that inhibit CYP3A4 selectively, pharmaceutical compositions, and methods of using the compounds and composition for treating a disease or disorder with a drug that is a P450 enzyme substrate. In addition, the disclosed compounds and compositions can be used for preventing P450 enzyme metabolism of a drug. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present invention.

The invention relates to gene modification, in particular to a method for improving the catalytic capacity of engineering P450 peroxygenase hydrogen peroxide and application of the method. The invention relates to a method for improving hydrogen peroxide catalytic capacity of P450 enzyme, which comprises the following steps of: identifying a channel of the P450 enzyme by using Caver Web 1.0 software by taking a heme cofactor of the P450 enzyme as a starting point; the method comprises the following steps: selecting a target channel with the channel length of 10-30, the curvature of 0-2 and the diameter of 0-3 as a target channel, then selecting amino acid of which the side chain faces the channel at an inlet or a bottleneck of the target channel as key amino acid, and then performing specific mutation on one or more key amino acids, further, at least one channel capable of improving the hydrogen peroxide catalytic capability of the P450 enzyme is obtained, so that the catalytic capability of the P450 enzyme is improved under the catalysis of hydrogen peroxide. The catalytic activity of a plurality of mutants obtained by the invention is compared with that of a wild enzyme, and the channel with high catalytic activity is an H2O2 channel. The application of the method can improve the hydrogen peroxide utilization capability of the P450 enzyme, and further improves the activity of the catalytic substrate of the P450 enzyme.

The present invention discloses a P450 enzyme mutant, a single-plasmid three-enzyme co-expression system, and their application in calcifediol synthesis. The P450 enzyme mutants include: using the wild-type P450 enzyme CYP109E1 shown in SEQ ID NO. 1 as a template, and respectively mutating the T at position 78 or the G at position 81 to form the mutants T78A, T78V, T78L, and T78I, G81V, and G81M; and mutating both the T at position 78 and the G at position 81 to form the mutant T78L / G81M. The present invention also constructs a single-plasmid three-enzyme co-expression system of the P450 enzyme mutant and a redox partner, thereby improving the electron transfer efficiency in the P450 enzyme hydroxylation reaction. This system can synthesize calcifediol with higher efficiency under mild conditions, solving the problems of low catalytic efficiency and activity in the synthesis of calcifediol catalyzed by P450 enzymes, and has important industrial application value.

The invention aims to catalyze bufalin and resibufogenin to modify and synthesize bufadienolide compounds, and discloses five newly discovered bufo bufo gargarizans source P450 enzymes, enzyme compositions, nucleic acids, expression vectors, bioengineering strains, methods and applications. The invention also discloses reported enzymes and enzyme compositions, nucleic acids, expression vectors, bioengineering strains, methods and applications of the P450 enzyme Sth10 related to catalysis of bufalin and resibufogenin for modification and synthesis of bufadienolide compounds. The amino acid sequence of the P450 enzyme is as shown in SEQ ID NO. 1-6. According to the invention, a plurality of P450 enzymes capable of modifying bufalin and / or resibufogenin at a fixed point under a mild condition are discovered for the first time, and a bioengineering strain and a biocatalytic synthesis method are developed based on the enzymes, so that a novel biosynthesis method for synthesizing bufadienolide compounds is established.

The invention belongs to the technical field of biochemical engineering, and particularly relates to application of a P450 enzyme mutant and engineering bacteria thereof in synthesis of calcifediol, and the mutant is obtained by mutating one or more sites in an amino acid sequence based on a wild type P450 enzyme Vdh; the amino acid sequence of the wild type P450 enzyme Vdh corresponds to the accession number of GenBank, namely CP069288.1. Through systematic molecular dynamics simulation, conservative analysis and substrate binding pocket engineering on the Vdh enzyme, a plurality of key mutation sites are successfully screened and obtained. The constructed combined mutant (such as I144R / N173M / Q310R, Vdh-M3) shows extremely high hydroxylation activity, the pure enzyme activity of the combined mutant can reach 9.8 times that of a wild type enzyme, and the conversion rate from vitamin D3 to calcifediol is greatly increased.

The present application relates to the technical field of biosynthesis, and specifically discloses a method for selectively hydroxylating 16-position of dehydroabietyl acid based on CYP153 family cytochrome P450 enzyme, which comprises the following steps: plasmid construction, heterologous expression of CYP153A99-RhFRED in Escherichia coli, heterologous expression of opt13 in Escherichia coli, and crude enzyme catalytic conversion reaction. By optimizing the codon bias of the protein sequence of cytochrome monooxygenase in the CYP153 family, the p450 enzyme CYP153A99 with the biological catalytic function of catalyzing the hydroxylation of C16 position of dehydroabietyl acid is obtained, high-site selective oxidation of dehydroabietyl acid is realized, the substrate spectrum and application field of the CYP153 family P450 enzyme are expanded, and a new technical path is provided for the green synthesis of dehydroabietyl acid derivatives.

The present disclosure relates to relates to compounds that inhibit CYP3A4 selectively, pharmaceutical compositions, and methods of using the compounds and composition for treating a disease or disorder with a drug that is a P450 enzyme substrate. In addition, the disclosed compounds and compositions can be used for preventing P450 enzyme metabolism of a drug. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present invention.

The present invention relates to a synthetic gene cluster of an enfumafungin antibiotic and a synthetic method therefor. Specifically, in the present invention, key functional genes in the linking of a β-D-glucopyranose at position C3 of a fernane-type framework, the oxidation at position C2 into α-OH, the oxidative cleavage of ring E at C19-C20, and the acetylation of hydroxyl at position C2 during the biosynthesis of an enfumafungin antibiotic, i.e. fuscoatroside, are isolated, wherein the genes are named fsoA, fsoD, fsoE and fsoF, respectively. The present invention also provides encoding polypeptides thereof. Provided in the present invention are an artificial fusion enzyme gene, i.e. efuA(TC)fsoA(GT); and on the basis of the artificial fusion enzyme gene, the heterologous expression of four genes, i.e. efuA(TC)fsoA(GT), fsoD, fsoE and fsoF, can synthesize an enfumafungin precursor (13). The present invention clarifies an FsoE-mediated C-C bond breaking function of a P450 enzyme, and provides a key catalytically active residue of FsoE. The present invention reports the biosynthetic pathway of such compound for the first time, and establishes an important foundation for the green and efficient synthesis of the compound.

The invention provides a P450 enzyme CYP5103-BM capable of catalyzing 14 alpha-hydroxylation of progesterone, and a mutant CYP5103-BM-Y291H of the P450 enzyme CYP5103-BM. Wherein the gene sequence of the P450 enzyme CYP5103-BM is as shown in SEQ ID NO: 1, and the gene sequence of the mutant CYP5103-BM-Y291H is as shown in SEQ ID NO: 3. The transformants respectively carrying the CYP5103-BM gene and the CYP5103-BM-Y291H gene can be subjected to a whole-cell biological catalytic reaction to convert a substrate progesterone into 14 alpha-hydroxyprogesterone, and the conversion rates in 36 hours respectively reach 50.04% and 88.76%. The mutant CYP5103-BM-Y291H has higher catalytic activity on progesterone, the production cost of 14 alpha-hydroxyprogesterone can be greatly reduced, the production time can be shortened, and the mutant CYP5103-BM-Y291H has extremely high application value for industrial production of progesterone and research of C14-hydroxylated steroid drugs.

The invention discloses a colletotrichum linum P450 enzyme mutant and application thereof, the mutant is obtained by mutating 121st aspartic acid of an amino acid sequence of P450 enzyme CYP68J derived from a colletotrichum linum ST-1 strain into histidine, and the amino acid sequence is shown as SEQ ID NO: 1. Compared with a wild type P450 enzyme, the P450 enzyme mutant disclosed by the invention has the advantages that dehydroepiandrosterone can be efficiently converted into 7 alpha-hydroxy-dehydroepiandrosterone, and the ratio of 7 alpha-hydroxy-dehydroepiandrosterone to 7 alpha, 15 alpha-dihydroxy-dehydroepiandrosterone in a hydroxylation product can be increased from about 4: 5 to about 14: 1; an important way is provided for the biological conversion of the 7alpha-hydroxyl-dehydroepiandrosterone, and the method has important significance on the industrial production of the 7alpha-hydroxyl-dehydroepiandrosterone.

The application discloses an engineered yeast strain for expressing cytochrome P450 and application thereof, and belongs to the technical field of bioengineering. The application constructs a method for constructing a Saccharomyces cerevisiae platform strain for improving cytochrome P450 expression by using a Saccharomyces cerevisiae MAGIC whole genome screening library, uses a yeast strain containing a betalain biosensor and a Saccharomyces cerevisiae MAGIC whole genome screening gRNA library, and successfully screens a target gene capable of improving CYP76AD1 expression in Saccharomyces cerevisiae by means of high-throughput screening of whole genome perturbation and fluorescence coupling with the aid of flow cytometry. Meanwhile, the microenvironment reconstruction target genes are applied to improve the functional expression of other P450 enzymes, the universality is proved, and it is proved that the method can effectively establish a platform engineered yeast strain for high-activity expression of various P450 enzymes.

The application provides a P450 enzyme mutant, a fusion protein and application thereof, the mutant is selected from any one of the following: a mutant formed by mutating an amino acid at the 85th position of a P450 enzyme shown in SEQ ID NO: 1 into glycine, alanine or cysteine, a mutant formed by mutating an amino acid at the 89th position of a P450 enzyme shown in SEQ ID NO: 2 into glycine, a mutant formed by mutating an amino acid at the 93rd position of a P450 enzyme shown in SEQ ID NO: 3 into glycine, a mutant formed by mutating an amino acid at the 85th position of a P450 enzyme shown in SEQ ID NO: 4 into glycine, and a mutant formed by mutating an amino acid at the 89th position of a P450 enzyme shown in SEQ ID NO: 5 into glycine. The P450 enzyme mutant can catalyze intramolecular C-S bond formation, and through improving the product selectivity of the P450 enzyme, specific synthesis of high-purity sulfur-containing compounds is realized.

A strain of Yarrowia lipolytica was engineered to overexpress β-hydroxy β-methylglutaryl-CoA reductase and farnesyl pyrophosphate synthase, as rate-limiting enzymes in the mevalonate and sesquiterpenoid synthesis pathways respectively. Metabolite extracts from this strain were run on LC-MS and showed a number of novel compounds being produced, including terpenoids varying in lengths and oxidation states. Upon NMR and MS / MS structure validation as well as biochemical assays, these compounds were determined as a new class of non-natural compounds, bifunctional terpenoids. Studies on the overexpression of P450 enzymes, alcohol oxidase, aldehyde dehydrogenase, and alcohol dehydrogenase showed that expression of these enzymes in addition to β-hydroxy β-methylglutaryl-CoA reductase and farnesyl pyrophosphate synthase increase the production of bifunctional terpenoids. Bioactivity assays demonstrate the application of bifunctional terpenoids.

This invention discloses sterol side-chain cleaving enzymes and 3β-hydroxysteroid dehydrogenases / Δ from different sources. 4,5 Amino acid sequences, coding sequences, and applications of isomerases. This invention, based on synthetic biology and molecular pharmacognosy, explores sterol side-chain cleaving enzymes and 3β-hydroxysteroid dehydrogenases / Δ from different plant sources. 4,5 Isomerases, which will yield sterol side-chain cleavage enzymes and 3β-hydroxysteroid dehydrogenases / Δ 4,5 The coding sequences of the isomerases were constructed into expression cassettes, and using Saccharomyces cerevisiae as the chassis strain, highly efficient pregnenolone or progesterone synthesizing strains were obtained. The pregnenolone yield of the constructed strains reached as high as 14.46±2.16 mg / L, and the progesterone yield reached as high as 32.78±1.84 mg / L. This achieved high-level synthesis of pregnenolone and progesterone by genetic engineering methods, providing an important foundation for the green synthesis of steroid hormones.

The invention belongs to the technical field of gene engineering and microbial fermentation, and particularly discloses a cytochrome P450 reductase coding gene CPR2 from aspergillus nidulans and application thereof. The nucleotide sequence of the gene is shown as SEQ ID NO: 1, and the amino acid sequence of the protein coded by the gene is shown as SEQ ID NO: 2. According to the invention, the CPR2 gene (OECPR2) is identified as an optimal electron transfer regulatory element by performing function screening on eight CPR genes annotated in a genome, and the CPR2 gene is over-expressed in an echinocandin B production strain aspergillus nidulans, so that the intracellular electron transfer efficiency is remarkably enhanced, the enzyme catalytic activity of cytochrome P450 is driven, and the cytochrome P450 can be used for preparing an echinocandin B cell. And finally, the yield of echinocandin B is efficiently increased. Experiments show that the engineering strain for over-expression of CPR2 can enable the yield of echinocandin B to reach 1869.97 + / -96.98 mg / L in shake flask fermentation, which is 24.37% higher than that of the original strain. The invention provides a key gene element and an engineering strain for industrial efficient production of echinocandin B, and has important economic value.

The invention relates to the technical field of agricultural biology, in particular to a P450 enzyme mutant, a method for improving C3'hydroxylation activity of P450 enzyme and a method for generating 3 '-ODI through whole cells. The cytochrome P450 enzyme mutant disclosed by the invention can selectively catalyze the daidzein to generate a C3'hydroxylation reaction to generate corresponding 3 '-ODI, has excellent selectivity and relatively high yield, and provides an efficient and reliable catalytic tool for the C3' hydroxylation reaction of the daidzein. The P450 enzyme mutant, a glucose dehydrogenase driven NAD (P) H regeneration module and a redox partner system Fdx / FdR are subjected to multi-enzyme co-expression, a cofactor self-circulating single plasmid four-enzyme co-expression system is successfully constructed, and the method is mild in reaction condition and low in catalyst cost and is an ideal scheme for preparing 3 '-ODI.

The application discloses a P450 enzyme mutant, a coding gene and application thereof in preparation of cortisone, and belongs to the technical field of bioengineering. The P450 enzyme mutant is obtained by amino acid mutation of CYP11B1 from Homo sapiens with an amino acid sequence shown as SEQ ID NO. 1, and the amino acid mutation site is at least one of S146V, H331D and L440F. The P450 enzyme mutant provided by the application has a higher soluble expression level in Escherichia coli than wild-type CYP11B1, the catalytic level of the constructed recombinant expression strain is higher, 11-deoxycortisol can be used as a substrate to prepare cortisone, the yield of the product is high, no by-product is generated, and the P450 enzyme mutant has a good industrial application prospect.

The application discloses a kind of P450 enzymes and mutants thereof degrading phenoxy carboxylic acid herbicides, the P450 enzyme is respectively named as P450 BSβ , OleT JE ;The P450 enzyme mutant is respectively named as P450 BSβ -F46A, P450 BSβ -F79A, P450 BSβ -F173A, P450 BSβ -F289A, P450 BSβ -F292A;Its amino acid sequence is respectively shown as SEQ ID NO.1-7.The application also discloses the application of the P450 enzyme and mutants thereof in degrading phenoxy carboxylic acid herbicides and related genes in cultivating transgenic crops resistant to phenoxy carboxylic acid herbicides.Experiments prove that the efficiency of the P450 enzyme mutant in degrading phenoxy carboxylic acid herbicides can be up to 12.3 times of P450 BSβ , and the total conversion number of catalyzing 2,4-dichlorophenoxybutyric acid can be up to 8779.The P450 enzyme mutant disclosed in the application has the advantages of high catalytic efficiency, high protein expression and low industrial cost, and the whole operation process of degrading herbicides is simple, the process is mature, the cost is low, and has wide application prospect.

The invention discloses an engineering bacterium for converting 4-hydroxyphenyllactic acid to generate tanshinol, which is characterized in that a P450 enzyme subfamily gene CYP98A75 gene from a plant salvia miltiorrhiza is introduced into escherichia coli, and a P450 reductase gene CPR from the plant salvia miltiorrhiza is co-expressed to construct an escherichia coli recombinant engineering bacterium. The engineering bacterium can catalyze a substrate 4-hydroxyphenyllactic acid to generate tanshinol. The engineering bacterium obtained by the invention is simple in culture method, high in pronucleus yield, single in product and easy to purify, and has important industrial application value.

This invention belongs to the field of bioengineering technology and provides the application of P450 enzyme in catalyzing the oxidation of squalene. Several cytochrome P450 genes were cloned from the full-length transcriptome data of the medicinal plant *Alisma plantago-aquatica*. Preliminary screening of the gene sequences corresponding to the P450 enzyme was conducted using bioinformatics. Recombinant vectors were prepared using the screened genes and transformed into squalene-producing yeast for gene function verification. This demonstrated the oxidation effect of the P450 enzyme encoded by the AoCYP_3836 gene on squalene, providing a novel reaction mode different from the traditional 2,3-epoxidation reaction of squalene. The results show that the P450 enzyme encoded by the AoCYP_3836 gene can catalyze the oxidation of squalene to produce two new products: squalene-1,1'-diol and squalene-1-carboxylic acid-24-ol, providing a new pathway for the production of these squalene oxides.

The invention discloses a P450 enzyme CYP-SA and a mutant CYP-SA-MT3 of the P450 enzyme CYP-SA. The gene sequences of the P450 enzyme CYP-SA and the mutant CYP-SA-MT3 are respectively shown as SEQ ID NO: 1 and SEQ ID NO: 3. The transformant carrying the CYP-SA gene and the CYP-SA-MT3 gene can catalyze 17 alpha-hydroxyprogesterone to generate 11 alpha, 17 alpha-dihydroxyprogesterone, the 72-hour conversion rates respectively reach 54.3% and 86.4%, and the C11 alpha hydroxylation selectivity is respectively 82.4% and 93.2%. The P450 enzyme CYP-SA and the mutant CYP-SA-MT3 have an extremely high industrial application value in the production of the 11 alpha, 17 alpha-dihydroxyprogesterone by a biological method.

The application discloses a kind of self-sufficient P450 enzyme and mutant application catalyzing intermolecular C-S bond coupling, belong to enzyme engineering and organic synthesis field.The application is by directed evolution, the key active site of P450 enzyme is mutated, especially F424, V118 and C385 residue.The mutant constructed in the application significantly improves the activity of C-S bond coupling, especially V118A / C385H / F424P mutant is excellent on different aryl mercaptan substrate.The microbial cell expressing mutant can be used as whole-cell catalyst for C-S bond coupling reaction, and lays a solid foundation for industrial application.

This invention relates to the field of biotechnology, and more particularly to a P450 enzyme mutant for steroid C11α-hydroxylation, the amino acid sequence of which is shown in SEQ ID NO:2. In a whole-cell catalytic system of *Saccharomyces cerevisiae*, this P450 enzyme mutant achieved a 14-fold increase in the conversion rate of 17α-hydroxyprogesterone compared to the wild type. Expression of this mutant in a systematically modified *Pichia pastoris* chassis strain increased the substrate conversion rate to 96.5% and the product selectivity to 99.1%. Scale-up culture at 2.5 L, with a single feed of 60 g of substrate, yielded a 11α,17α-dihydroxyprogesterone titer of 24.8 g / L. Furthermore, this mutant also exhibited good catalytic activity and C11α-hydroxylation selectivity for various steroid drugs such as progesterone and canrenone, demonstrating a broad substrate spectrum and promising prospects for industrial applications.

The application discloses a cytochrome P450 enzyme derived from Stephania sinica Diels and application thereof in biosynthesis of aporphine alkaloid compounds. The application first characterizes a key P450 involved in formation of a methylenedioxy bridge structure of aporphine alkaloids in Stephania sinica Diels, provides an important reference for analysis of an aporphine alkaloid biosynthesis pathway in other species, and lays a foundation for heterologous production of various aporphine alkaloids. The application screens a P450 enzyme CYP719C3 annotated as CYP719 in a genome of Stephania sinica Diels, performs enzymatic reactions with different aporphine compounds as substrates, and detects reaction results by using a liquid chromatograph-mass spectrometer. It is found that CYP719C3 can catalyze an enzyme forming a methylenedioxy bridge on an A ring of an aporphine alkaloid, and can be applied to further analysis and heterologous production of an aporphine alkaloid pathway.

This invention belongs to the technical fields of genetic engineering and fermentation engineering, and discloses an animal cytochrome P450 oxidoreductase derived from coral and its applications. Its amino acid sequence is shown in SEQ ID NO:1, and the invention also relates to its encoding gene and recombinant vector. This invention integrates the CPR gene into the chromosome of *Saccharomyces cerevisiae* to construct a recombinant strain for driving P450 enzyme catalytic reactions. Experiments show that, compared to CPR from other sources, this coral-derived CPR significantly improves electron transport efficiency, effectively catalyzing the conversion of substrate into products 1 and 2. Under optimized fermentation conditions, the product conversion rate of this recombinant strain reaches 0.44, far exceeding that of plasmid expression systems. This invention provides an excellent electron transport element and engineered strain for the efficient application of P450 enzymes and the synthesis of natural products.

The invention relates to a synthetic gene cluster and a synthetic method of an amphetamine antibiotic. Specifically, key action genes for connecting beta-D-glucopyranose to the C3 site, oxidizing the C2 site into alpha-OH, oxidizing and cracking the E ring C19-C20 and acetylating the C2 site hydroxyl in biosynthesis of the amphetamine gold antibiotic fuscoatroside are separated and are respectively named as fsoA, fsoD, fsoE and fsoF, and the invention provides a coding polypeptide of the key action genes and the application of the key action genes in biosynthesis of the amphetamine gold antibiotic fuscoatroside in biosynthesis of the amphetamine gold antibiotic fuscoatroside in biosynthesis of the amphetamine gold antibiotic fuscoatroside in biosynthesis of the amphetamine gold antibiotic fuscoatroside in biosynthesis of the amphetamine gold antibiotic fuscoatroside. The invention provides an artificial fusion enzyme gene efuA (TC) fsoA (GT), and on the basis of the artificial fusion enzyme gene efuA (TC) fsoA (GT), four genes of efuA (TC) fsoA (GT), fsoD, fsoE and fsoF are subjected to heterologous expression, so that an ampelopsin precursor (13) can be synthesized. The invention clarifies a P450 enzyme FsoE mediated C-C bond breaking function, and provides a key catalytic activity residue of FsoE. The invention reports the biosynthesis pathway of the compound for the first time, and lays an important foundation for realizing green and efficient synthesis of the compound.