106 results about "Shake flask" patented technology

The invention relates to the technical field of genetic engineering, and particularly discloses a genetic engineering bacterium for producing melanin as well as a construction method and application of the genetic engineering bacterium. According to the application, the yield of the melanin can be effectively increased by modifying a metabolic pathway of the genetically engineered bacterium, and the yield of the melanin reaches 6.2 g / L and 18.5 g / L respectively through shake-flask culture and 5L fermentation tank culture. Compared with the prior art, the yield of melanin produced by the genetically engineered bacterium is improved by 30% at the shake flask fermentation level, the fermentation yield of a fermentation tank is improved by 4.3 times, in addition, expensive tyrosine is changed into glucose as a fermentation precursor substance, the production cost is greatly reduced, and an efficient and feasible solution is provided for large-scale production of melanin.

The invention discloses an aspergillus niger recombinant bacterium AnCT-XynB-K-agdA and application thereof, and belongs to the technical field of microbiology and bioengineering. According to the invention, Aspergillus niger AnCat is taken as an expression host, firstly, an acid protease regulatory factor prtT is knocked out to obtain a defective strain, and then different chaperonins are subjected to fusion expression by optimizing integration sites of a laccase lcc9 expression cassette, so that the high-expression laccase chaperonin is obtained. According to the method, an expression vector pC3-5 'agdA-PcitA-XynB-L-lcc9-hph-3' agdA is constructed, a recombinant bacterium AnCT-XynB-K-agdA is further obtained, the enzyme activity of the extracellular laccase subjected to shake flask fermentation reaches 1821.2 U / L, and finally, the recombinant efficient expression of the laccase Lcc9 in aspergillus niger is realized.

The invention discloses a construction method of nicotinamide engineering bacteria for cosmetics, which comprises the following steps: amplifying target DNA, recovering the target DNA, carrying out seamless cloning or CPEC connection on the recovered DNA fragment and a vector, carrying out seamless cloning by using a seamless cloning kit of an all-type gold company, carrying out connection by using a sterile ultrapure water complementing system, and carrying out plasmid construction. The method comprises the following steps: inoculating a liquid culture medium with a strain which is verified to be correct, carrying out plasmid extraction on the obtained bacterial liquid by using an AxyPrep plasmid small extraction kit, carrying out shake flask small test on enzyme activity, thallus propagation and protein expression after chassis strain gene editing, adding a substrate 3-cyanopyridine solution into the crushed thallus for reaction, and carrying out HPLC (High Performance Liquid Chromatography) detection on nicotinamide. According to the method, the nitrile hydratase is expressed through metabolism of microorganisms, conversion from 3-cyanopyridine to nicotinamide is achieved through one-step reaction after cell disruption, the enzyme activity of the nitrile hydratase is improved, and meanwhile the generation amount of the by-product nicotinic acid is reduced.

The invention relates to the technical field of microbial directional cultivation, in particular to a gene editing-based MICP functional bacteria directional cultivation method and system, and the method comprises the following steps: obtaining an original strain library with MICP activity, carrying out strain activation on the original strain library, carrying out high-throughput sequencing on the original strain library, and carrying out gene mining on an initial sequence; the method comprises the following steps: carrying out gene editing on strains in an initial strain library by using a gene editing tool, carrying out shake-flask activation on engineered candidate strains to obtain an activated recombinant bacterium solution, carrying out high-density culture on the activated recombinant bacterium solution by using a pre-constructed fermentation tank, screening out an MICP strain with the optimal function, carrying out continuous passage bacterium function detection on the optimal MICP strain, and carrying out high-density culture on the MICP strain with the optimal function. And based on the MICP functional bacteria, finishing directional cultivation of the MICP functional bacteria based on gene editing. According to the invention, directional cultivation of the MICP functional bacteria is efficiently realized by accurately targeting the key gene with the MICP function, and the MICP functional bacteria are corrected when the problems of function instability and the like exist, so that the more stable MICP functional bacteria are obtained.

The invention relates to the technical field of synthetic biology, in particular to a squalene high-yield saccharomyces cerevisiae engineering bacterium and a construction method and application thereof.Saccharomyces cerevisiae CEN.PK.2-1D is used as an original strain, pentose phosphate pathway genes ZWF1, GND1, TAL1 and TKL1 and IDP1 genes of TCA circulation are integrated in a genome of the original strain through homologous recombination, and the squalene high-yield saccharomyces cerevisiae engineering bacterium is obtained. Constructing a dual-channel cofactor supply system to improve the intracellular NADPH level and the NADPH / NADP + ratio; according to the saccharomyces cerevisiae genetic engineering strain Sc-Sq04 constructed by the method disclosed by the invention, the yield of squalene obtained by shake flask fermentation reaches 741.08 + / -28.9 mg / L by further integrating all genes of an MVA way, ERG20 and ERG9 genes, and a new thought is provided for transforming saccharomyces cerevisiae to synthesize terpene compounds and promoting industrial application.

The invention provides pichia pastoris capable of efficiently expressing TFF3 fusion protein as well as a construction method and application of the pichia pastoris, and belongs to the technical field of engineering bacteria. The preservation number of the pichia pastoris is CCTCC (China Center For Type Culture Collection) NO: M 2025013. The pichia pastoris constructed by the invention can obviously improve the expression quantity of the TFF3, the shake flask expression quantity is up to 391 mg / L, and the pichia pastoris has a wide application prospect in large-scale industrial production of the TFF3.

The invention discloses recombinant poria cocos chitin endonuclease as well as a gene, a preparation method and application thereof. The nucleotide sequence of the gene is shown as SEQ ID NO.1 in a sequence table. According to the nucleotide sequence as shown in SEQ ID NO.1, high-level recombinant secretory expression of target protein in pichia pastoris host bacteria can be realized by taking pichia pastoris inducible expression plasmid pPICZ alpha A as a carrier. Under a shake flask fermentation condition, the average secretory expression quantity can reach 300.3 mg / L. The recombinant enzyme obtained by nickel ion affinity chromatography purification not only can effectively hydrolyze colloidal chitin to generate chitosan oligosaccharide (COS), but also can directly degrade chitin-containing raw materials such as shrimps, crabs and the like to generate COS. In addition, the enzyme also shows the activity of hydrolyzing polysaccharides such as cellulose, pectin, agar and the like, and has important potential values in the fields of functional chitosan oligosaccharide preparation, biological energy production, medicine development, feed, environmental protection, agricultural application and the like.

The invention discloses a stenotrophomonas sp. A322 strain and an application of the stenotrophomonas sp. A322 strain in synthesis of acarbose. The stenotrophomonas sp. A322 is preserved in the China Center for Type Culture Collection, and the preservation number of the stenotrophomonas sp. A322 is CCTCC (China Center for Type Culture Collection) NO: M 2025917. The stenotrophomonas sp. A322 can be used for synthesizing acarbose by using maltose and glucose as carbon sources, and the acarbose can be synthesized by using the stenotrophomonas sp. The stenotrophomonas sp. A322 is subjected to fermentation culture in a 100 mL shake flask fermentation system for 1 to 7 days, and the acarbose yield is 0.4 to 4 g / L. The high performance liquid chromatography retention time and the nuclear magnetic resonance carbon spectrum chemical shift of the acarbose obtained by fermentation, separation and purification of the Stenotrophomonas sp. A322 are consistent with those of a standard substance, and the high performance liquid chromatography retention time and the nuclear magnetic resonance carbon spectrum chemical shift of the acarbose obtained by fermentation, separation and purification of the Stenotrophomonas sp. A322 are consistent with those of the standard substance. Activity detection shows that the separated and purified acarbose sample has alpha-amylase inhibitory activity.

The invention provides a method for producing L-carnosine, a genetic engineering strain as well as a preparation method and application of the genetic engineering strain, and belongs to the field of genetic engineering. According to the genetic engineering strain provided by the invention, by knocking out a glucose-6-phosphate isomerase coding gene pgi, knocking out an L-threonine / L-homoserine transporter coding gene rhtA or knocking out a dipeptide transfer protein coding gene dppABCD, the yield of extracellular L-carnosine is increased, the shake flask yield of the finally constructed strain reaches 9.1 g / L, the 5L fermentation tank yield reaches 65 g / L, and the yield of the extracellular L-carnosine reaches 9.1 g / L; and the method has a good application prospect in L-carnosine biosynthesis.

The invention provides a beta-glucosidase mutant from Paenibacillus phyllosphaerae, and a method for efficiently preparing rare ginsenoside F2 by using an enzyme method and a whole-cell method, and particularly provides a beta-glucosidase mutant from Paenibacillus phyllosphaerae and a method for efficiently preparing rare ginsenoside F2 by using a beta-glucosidase mutant from Paenibacillus phyllosphaerae. The method comprises the following steps: engineering-modifying beta-glucosidase by utilizing computer-aided design, constructing a mutant library, carrying out primary screening and secondary screening to obtain two high-activity mutants Q7D and G189A, constructing a combined mutant Q7D / G189A, transforming the combined mutant Q7D / G189A into genetically engineered bacterium escherichia coli for expression, and producing F2 by utilizing a whole-cell method. Finally, the highest yield of 18.01 g / L is achieved after the reaction is carried out for 10 h in a 5 L fermentation tank and is 6.82 times that of a shake flask, and the purity of F2 reaches up to 82.20% after the reaction is carried out for 24 h finally. The invention not only provides an efficient solution for large-scale production of rare saponin F2, but also provides a transferable framework for designing and engineering diversified glycoside hydrolase in customized biological catalysis.

The invention discloses a pinene synthase mutant and application thereof in pinene production, and belongs to the technical field of bioengineering. Isoleucine at the 409th site of pinene synthase is mutated into valine, phenylalanine at the 443rd site of pinene synthase is mutated into alanine, overexpression is carried out in serratia marcescens (HBQA7), and an engineering strain ST17 is constructed. Under the condition of shake flask fermentation, the pinene yield of the engineering strain ST17 reaches 0.80 g / L; the yield is obviously increased to 43.2 g / L through amplification culture in a 30 L fermentation tank. The invention provides an efficient, economic and environment-friendly novel method for industrial biosynthesis of pinene.

1. Name of the product in this design: Packaging Bottle (Explosive Shake Bottle). 2. Purpose of this design: This product is a packaging bottle used to hold liquids. 3. The key design feature of this product is its shape. 4. The image or photograph that best illustrates the design's key points: a 3D model. 5. Other matters requiring explanation: Part A is made of transparent material. The white liquid in this design product is the contents and is not part of the design for which protection is sought.

This invention discloses a high-yield lactyl-3-4 ...5-4-4-4-4-4- N This invention relates to plasmid-free genetically engineered strains of tetrasaccharides and their applications, belonging to the field of microbial genetic engineering. The invention screened strains with higher activity. β -1,3-galactosyltransferase gene PmgalT Computer-aided enzyme function modification was used to achieve a shake-flask yield of 10.05 g / L for the constructed mutant. Subsequently, genome integration was performed... breast Multi-copy integration PmgalT * V27I / V39I / Y147F ‑galE‑galU A plasmid-free strain was constructed, enabling it to grow in shake flasks with lactoyl- N The yield of tetrasaccharides reached 14.4 g / L. Using glucose as the sole carbon source, the highest yield of 68.19 g / L was achieved at 64.5 h; using glycerol as the sole carbon source, the highest yield of 62.88 g / L was achieved at 60.5 h, providing a basis for its industrial production.

This invention discloses a nicotinamide phosphoribosyltransferase mutant, a recombinant strain, and a method for constructing the same. The mutants are I457V, G344A, Q378A, T283G, or F182A, obtained by single-point mutation of the wild-type nicotinamide phosphoribosyltransferase shown in SEQ ID NO:1. This invention utilizes a method derived from... Schizotequatrovirus By rationally designing and site-directed mutagenesis of KVP40 nicotinamide phosphoribosyltransferase, combined with metabolic engineering to block the NMN degradation pathway and enhance substrate synthesis and product transport, the yield of β-NMN by the recombinant strain during shake-flask fermentation reached 4.82 g / L. Through optimization of fed-batch fermentation conditions, the highest yield of β-NMN during fed-batch fermentation in shake flasks reached 15.60 g / L.

The invention discloses an escherichia coli strain with high yield of riboflavin as well as a construction method and application of the escherichia coli strain. A series of Escherichia coli strains with high yield of riboflavin are constructed, the constructed Escherichia coli with high yield of riboflavin is safe and harmless, glucose is used as a substrate for fermentation under the aerobic condition of a shake flask, the yield of riboflavin is 1117.8 mg / L, 1330.1 mg / L, 1430.8 mg / L, 1508.7 mg / L and 1640.8 mg / L in sequence, and the yields are 112.5 mg / g of glucose, 134.1 mg / g of glucose, 146.3 mg / g of glucose, 158.8 mg / g of glucose and 168.1 mg / g of glucose in sequence; when the strain SN04M is subjected to fed-batch fermentation in a 1L fermentation tank, the yield of riboflavin reaches 8.52 g / L, and the total yield is 189.9 mg / g glucose. According to the invention, the fermentation yield of riboflavin and the substrate utilization rate of riboflavin to glucose can be obviously improved.

The invention relates to pichia pastoris with high yield of Brazilian sweet protein and application of the pichia pastoris, and belongs to the technical field of fermentation. The invention finds that two molecular chaperones ERV29 and CPR5 from saccharomyces cerevisiae jointly strengthen the expression of the Brazilian sweet protein so as to significantly improve the yield of the Brazilian sweet protein, the Pichia pastoris provided by the invention is subjected to fermentation culture, the yield of the Brazilian sweet protein in a shake flask reaches 1.8 g / L, the yield of the Brazilian sweet protein in a 5-L fermentation tank reaches 11.3 g / L, which is the highest yield of the Brazilian sweet protein in the existing strain, and the Pichia pastoris provided by the invention has a wide application prospect. The method has a wide application prospect in industrial production of Brazilian sweet protein.

This invention relates to the field of fungal strain cultivation technology, and particularly to a method for isolating and cultivating fungal strains from matsutake fruiting bodies. The method involves collecting matsutake mushrooms that are 70%–80% mature and free from insects, disinfecting them with alcohol, longitudinally splitting them in half, removing the flesh from the cap and stipe, and inoculating them onto a slant culture medium to obtain a primary strain. This primary strain is then inoculated into a culture medium and shaken to obtain a primary shake flask strain, which is then inoculated into a culture medium. The strain is then shaken and cultured, and the concentration of metabolites, dissolved oxygen, and pH in the culture medium are collected in real time to construct a strain growth time prediction model to predict the stage culture duration. The cell proliferation trend is determined based on the number of matsutake mycelial cells. Based on a comparison of the actual culture time during the initial adjustment period and the stage culture time, the adjustment method for the logarithmic growth phase of the current batch is determined. Based on a comparison of the shake flask culture time and the predicted culture time, the adjustment method for the initial culture conditions of the next batch is determined. This invention effectively optimizes shake flask culture conditions and improves strain quality.

The invention provides a bacillus high-yield strain for producing riboflavin and application of the bacillus high-yield strain, and belongs to the technical field of microbial fermentation. The bacillus strain is named as VB2-M-230602-66, the preservation unit is China Center for Type Culture Collection, the preservation date is December 1, 2023, and the preservation number is CCTCC NO: M20232421; according to the invention, the strain VB2-M-230602-66 with stable character is obtained by adopting physical mutagenesis separation and combining with shake flask screening of tolerance dose of a resistance marker, riboflavin can be produced through liquid fermentation, and the yield is stable; the bacterial strain has the biological characteristics that the bacterial strain is gram-positive bacteria through gram staining, the bacteria are cultured at 37 DEG C for 18 hours, the thalli are rod-shaped, 0.5-0.8 mu m * 2.0-5.4 mu m and are arranged in a single or paired manner, and bacterial colonies cultured on a solid culture medium are pale yellow, nearly circular, wet in surface, opaque and irregular in edge; the bacillus strain VB2-M-230602-66 provided by the invention is suitable for fermentation production of riboflavin (vitamin B2), has stable production performance, can obtain greater economic benefits, and is worthy of popularization.

The utility model relates to the technical field of cell culture, and discloses a cell shake flask capable of improving tissue digestion efficiency, which comprises a culture flask body, a flask cover is mounted at the top end of the culture flask body, and a first constant-temperature heating cover is arranged on one side of the outer side wall of the culture flask body. Mounting blocks are fixed to the front end and the rear end of one side of the first constant-temperature heating cover correspondingly, inner cavities are formed in the first constant-temperature heating cover and the second constant-temperature heating cover correspondingly, and electric heating wires are mounted in the inner cavities. When the culture bottle body disclosed by the utility model is used for culturing cells, the culture bottle body can be heated according to actual conditions by arranging components such as the first constant-temperature heating cover, the mounting block, the mounting groove and the second constant-temperature heating cover, and meanwhile, the digestion efficiency of cell tissues can be promoted by heating the culture bottle body; and the culture bottle body can be detached when not needed subsequently, so that the observation of the condition of cells in the culture bottle body through the culture bottle body is not influenced, and the use effect is better.

The invention relates to the field of scalp care, and particularly discloses a preparation method of a scalp and skin care preparation, which comprises the following steps: S1, a strain activation step: taking bacillus subtilis as a preparation strain, inoculating the strain in a preservation state into an LB agar culture medium with the pH value of 7, and performing 2-3 generations of culture rejuvenation to obtain vigor-rich strain seeds; s2, shake-flask culture: selecting the vigor-rich strain seeds, and inoculating the vigor-rich strain seeds into an LB liquid culture medium with the pH value of 7; culturing the inoculated LB liquid culture medium in a shaking table to obtain a shake flask bacterial liquid; s3, a fermentation culture step: firstly, preparing a fermentation culture medium, and adjusting the pH value of the fermentation culture medium to 7; then carrying out fermentation operation; according to the technical scheme, the product has high content of active components, the release efficiency of active substances of the bacillus subtilis preparation is high, and the final preparation has the safety, antibacterial and repairing capabilities.

This invention discloses a bifunctional flavin adenine dinucleotide synthase mutant and its application in FAD synthesis. The mutant disclosed in this invention is RFK / FADS. C18R or RFK / FADS C18R‑R46H ;RFK / FADS C18R The amino acid sequence is shown in SEQ ID NO.15, RFK / FADS C18R‑R46H The amino acid sequence is shown in SEQ ID NO. 16. Genetically engineered bacteria carrying the bifunctional flavin adenine dinucleotide synthase mutant gene plasmid produced flavin adenine dinucleotide using glucose as a substrate through a simple fermentation process under aerobic conditions in a shake flask. The FAD yields of strains XF02 and XF03 containing the mutant gene were 79.31 mg / L and 167.56 mg / L, respectively, representing increases of 58.62% and 235.12% compared to strain XF01 containing the non-mutated gene. By constructing microbial "cell factories" through metabolic engineering strategies, modified microorganisms can utilize inexpensive raw materials to generate desired natural products within their cells. This advancement not only reduces the production cost of FAD but also improves the environmental sustainability of its production process.

The invention belongs to the technical field of genetic engineering, and particularly relates to hidden plasmid mediated genetic engineering probiotics as well as a construction method and application thereof. According to the method, stable genetic expression of exogenous genes is mediated through high-copy hidden plasmids, resistance does not need to be added in the culture process, and the safety, stability and effectiveness of synthesis of food-grade nutritional chemicals are remarkably improved. The modified strain can efficiently utilize glucose as a carbon source to synthesize a functional nutritional chemical VB5 product, the titer of VB5 in shake flask fermentation reaches 102 mg / L, and the product is not detected by a wild strain. The invention provides a new strain and technical support for food-grade efficient synthesis of functional nutritional chemicals, and is beneficial to popularization and application in the field of food.

The utility model belongs to the technical field of microbial strain transfer, and discloses a shake flask strain transfer auxiliary device which comprises a supporting seat, a containing groove and a pressure bearing piece are arranged on the upper portion of the supporting seat, the pressure bearing piece is located on one side of the containing groove, a positioning groove used for fixing a shake flask neck is formed in the pressure bearing piece, and a shake flask is obliquely placed in the containing groove. And the bottleneck of the shake flask is embedded in the positioning groove. The shake flask support is used for supporting a shake flask which is obliquely placed, a traditional handheld shake flask mode is replaced, and the probability that the shake flask shakes and the hand is burnt by flames can be effectively reduced.

The invention discloses a recombinant genetically engineered bacterium for producing L-homoserine at high yield and application of the recombinant genetically engineered bacterium. L-homoserine transporter with higher efficiency is obtained by editing, superposing and screening multiple genes, and the yield of L-homoserine is further increased by overexpressing thrE gene; and screening to obtain the recombinant genetically engineered bacterium capable of improving the yield of the L-homoserine. The yield of the L-homoserine in a shake flask reaches 5.62 g / L, and compared with a control strain, the yield of the L-homoserine is improved by 43%; the yield of the L-homoserine in a 5L fermentation tank reaches 30.9 g / L within 72 hours, and compared with a control strain, the yield of the L-homoserine is improved by 72.2%.

The invention discloses a protein fermentation, purification and preparation process of acinetobacter baumannii vaccine antigen Ata fragment protein. The process comprises the following steps: S1, carrying out inoculated culture through a shake flask; s2, thalli are collected; s3, crushing the thalli, and carrying out centrifugal clarification; s4, anion chromatography; s5, hydrophobic chromatography; s6, desalting and changing liquid; and S7, obtaining the high-purity Ata fragment protein. According to the method, a process from small-scale strain fermentation to separation and purification is established, a protein sample with the yield of-g level is finally obtained by combining anion exchange, hydrophobic chromatography, ultrafiltration and other methods, and the protein purity reaches 99%.

The application discloses a method for improving cellulase activity by using a combination of an endogenous promoter and an associated 5' UTR of Trichoderma reesei and an engineering strain constructed by the method. TrAE The 5' UTR upstream of the TrAE gene TrAE Overexpressing a transcriptional regulator xyr1 The expression cassette of the gene is transformed into Trichoderma reesei Δ ku70 Rut‑C30 The application also discloses a high-yield cellulase Trichoderma reesei strain QEaeuX, and a method for producing high-activity cellulase by using the strain. ku70 Rut‑C30 The cellulase activity produced by the strain in a 96 h shake flask culture is 2.6 IU / mL, which is 101% higher than that of the original strain Δ The application has a good industrial development and application prospect.

This invention discloses a method for constructing a high-succinic acid-producing Pichia pastoris cell factory, using Pichia pastoris (… Komagataella phaffii Using GS115 as the chassis, a reductive TCA cycle enhancement strategy and an sdh5 knockout strategy were first constructed, respectively. It was found that a single strategy was insufficient to effectively achieve succinic acid accumulation. Further, the two strategies were combined to obtain the engineered strain SAOG, which achieved succinic acid accumulation. Based on this, the malate transporter SpMAE was introduced to enhance the transmembrane transport capacity of succinic acid, and histidine auxotrophic recovery was combined to improve the growth and fermentation performance of the strain, resulting in the engineered strain SH, whose succinic acid accumulation was 25.1% higher than that of SAOG. Finally, the engineered strain SH achieved a succinic acid yield of 10.47 g / L in shake-flask fermentation and 112.6 g / L after 120 h of continuous fed-batch fermentation in a 5 L fermenter. This invention achieves efficient succinic acid synthesis in the Pichia pastoris chassis through the synergistic regulation of reductive TCA pathway enhancement, succinic acid oxidation blocking, enhanced transmembrane transport, and auxotrophic recovery, exhibiting stable fermentation performance, high yield, and good industrial application potential.

The invention provides pichia pastoris for producing a honey truffle sweetening agent as well as a construction method and application thereof, and relates to the technical field of microorganisms. According to the invention, high-efficiency expression of a natural sweet protein honey truffle sweetening agent is realized in pichia pastoris cell engineering, a screened signal peptide C terminal is connected with a Pro region of an alpha mating factor to obtain a fusion signal peptide, and heterologous expression of a honey truffle sweetening agent coding gene is realized through the fusion signal peptide. The pichia pastoris capable of efficiently secreting and expressing the target sweetening agent in an extracellular mode is successfully constructed, the pichia pastoris can be used for efficiently producing the honey truffle sweetening agent, the shake flask yield reaches 44 mg / L, and an efficient and feasible new strategy is provided for large-scale microbial production of the honey truffle sweetening agent.

The application provides a recombinant Aspergillus niger strain with high trans-aconitic acid yield, and belongs to the technical field of genetic engineering. The recombinant Aspergillus niger strain can simultaneously express aconitase and aconitate isomerase. The genetically engineered Aspergillus niger strain provided by the application has a generally higher trans-aconitic acid yield than a starting strain, and the strain with the highest yield can reach 51 g / L at a shake flask level, and has a very strong application value.

The application discloses an engineered strain for synthesizing resveratrol by using p-coumaric acid as a substrate, construction and application thereof, and belongs to the technical field of biotechnology. The active sites of 4-coumaroyl-CoA ligase and resveratrol synthase are semi-rationally designed and modified, the optimal mutants 4CL1 (M4) and VST1 (N3) are expressed in a Yarrowia lipolytica strain, and the obtained engineered strain can efficiently convert p-coumaric acid into resveratrol. The yield of resveratrol in a shake flask fermentation reaches 724.6 mg / L. Then, the two mutants are fused and expressed by using a rigid linker, and the Yarrowia lipolytica engineered strain for producing resveratrol by using p-coumaric acid as a substrate is obtained. In a 5-L fermenter, batch feeding fermentation is carried out, the substrate conversion rate reaches 92.3%, and the yield of resveratrol reaches 35.8 g / L.