965results about "Lyases" patented technology

The invention discloses a mesophilic carotenoid-producing recombinant yarrowia lipolytica strain as well as a construction method and application thereof, the recombinant strain exogenously expresses mesophilic original fatty acid desaturase (FAD3) and isocitrate dehydrogenase (IDH2) mutants from phaffia rhodozyma, and efficient synthesis under the condition of 30 DEG C is realized; secondly, a metabolic pathway of the recombinant strain is optimized based on a metabolic flow push-pull strategy, and isopentenyl pyrophosphate isomerase (IDI), farnesyl pyrophosphate synthase (ERG20), plant alkene dehydrogenase (CarB) and bifunctional plant alkene synthase (CarRP) are overexpressed, so that the flux of a precursor is increased; finally, gene multi-copy of astaxanthin synthase (crtS) and cytochrome P450 reductase (crtR) from phaffia rhodozyma is carried out, and the carotenoid production capacity of yarrowia lipolytica is further improved. The construction method of the mesophilic yarrowia lipolytica is simple, efficient synthesis of carotenoid can be better achieved at the higher temperature, the fermentation cost is greatly reduced, and the industrial production potential is achieved.

The invention discloses an arginine decarboxylase diaA enzyme mutant and application thereof in preparation of butanediamine, and belongs to the field of bioengineering. According to the invention, rational charge overturning transformation is simultaneously carried out on a pentamer meridian oligomeric interface and a dimer latitudinal oligomeric interface, so that stable assembly and efficient catalysis of the decamer under the condition of neutral to alkaline pH (7.0-9.0) are realized. Wherein positive charges are introduced into a meridian interface to weaken electrostatic repulsion, and negative charges are introduced into a latitudinal interface to enhance dimer compactness and substrate transfer efficiency. The specific enzyme activity of the representative double mutant AdiAD110K / H736E at pH 8.0 is about 35 times that of a wild type, and the representative double mutant AdiAD110K / H736E keeps a complete decamer state in a pH range of 7.0-9.0. The yield of butanediamine is up to 145.9 g / L under the whole-cell catalysis of the mutant.

The invention discloses a rational copolymerization strategy for improving the stability of an oligomeric structure of acid-induced high-order oligomeric decarboxylase AdiA in a neutral to alkaline environment and application of the rational copolymerization strategy, and belongs to the field of bioengineering. According to the strategy, rational charge overturning transformation is carried out on a pentamer radial oligomeric interface and a dimer weft-wise oligomeric interface at the same time, and stable assembly and efficient catalysis of a decamer under the condition that the pH value is from 7.0 to 9.0 from neutral to alkaline are achieved. Wherein positive charges are introduced into a meridian interface to weaken electrostatic repulsion, and negative charges are introduced into a latitudinal interface to enhance dimer compactness and substrate transfer efficiency. The specific enzyme activity of the representative double mutant AdiAD471K / E467K / H736E is 45.5 times that of a wild type when the pH value is 8.0, and the representative double mutant AdiAD471K / E467K / H736E keeps a complete decamer state when the pH value is 7.0-9.0. The yield of butanediamine is up to 156.5 g / L by using the mutant to catalyze whole cells.

The invention discloses a high-efficiency lysine decarboxylase mutant and an application of the high-efficiency lysine decarboxylase mutant in synthesis of pentamethylene diamine. A series of mutants including a single mutant (such as N218G, T222V, C244P and the like), a double mutant and a three mutant are obtained by carrying out multi-site mutation on wild type lysine decarboxylase (SEQ ID NO.2) from Klebsiella grimontii. The catalytic efficiency of the mutant is remarkably improved, and the activity of the mutant is improved by 5.0-10.2 times compared with that of a wild type enzyme. According to the method disclosed by the invention, the substrate conversion rate of 99% or above can be realized within 2 hours through an optimized process of constructing an expression vector and a genetically engineered bacterium, inducing expression of lysine decarboxylase and synthesizing 1, 5-pentanediamine by utilizing a whole-cell catalysis technology. Compared with a traditional chemical method and a natural enzyme method, the method disclosed by the invention has the advantages of simplified process, high catalytic efficiency, low production cost and the like, and is suitable for producing bio-based 1, 5-pentamethylene diamine and nylon 56 salt.

The invention discloses a tryptophan enzyme mutant and application thereof. The tryptophan enzyme mutant is obtained by mutation of at least one site of the 50th amino acid L, the 394th amino acid V and the 395th amino acid E of a tryptophan enzyme parent. The tryptophan enzyme mutant disclosed by the invention has good substrate bias, particularly has better selectivity and catalytic efficiency on 6 '-halogenated tryptophan, and not only simplifies the production process, but also improves the production efficiency when being used for biosynthesis production of the tylosin.

The invention relates to the field of biological engineering and technology, in particular to an enzyme for synthesizing glabridin, a biological synthesis method of glabridin and a bacterial strain. The invention discloses a series of glabridin synthesizing enzymes (phenylalanine ammonialyase, cinnamic acid-4-hydrogenase, tyrosine ammonialyase, p-coumaric acid coenzyme A ligase, chalcone synthase, chalcone reductase, chalcone isomerase, isoflavone synthase, 2-hydroxyisoflavone dehydratase, 4 '-oxymethyltransferase, 4'-hydroxymethyltransferase and 4 '-hydroxymethyltransferase) derived from glycyrrhiza glabra for the first time. ) can be used in the preparation of a compound (e.g., isoflavone 2 '-hydrogenase, isoflavone reductase, Vissisoketoreductase, isoprenyltransferase, pterocarpin reductase, oxidative cyclase, demethylase, cytochrome P450 reductase, and the like). The de novo synthesis of the glabridin in a microbial cell factory is realized for the first time, and the application prospect of the glabridin biological manufacturing industry is greatly promoted.

The invention provides a kit for rapidly detecting MRSA and drug-resistant gene nucleic acid POCT of the MRSA based on a phage lyase mutant. The kit comprises nucleic acid extraction and purification and triple PCR rapid amplification detection reagents. The staphylococcus aureus bacteriophage lyase mutant nucleic acid release liquid containing the amino acid sequence as shown in SEQ ID NO: 1 is adopted, wall breaking can be rapidly conducted on staphylococcus aureus, and nucleic acid is released. The effect of the nucleic acid release liquid acting for 5 minutes is equivalent to or even better than that of the lysozyme acting for 60 minutes; according to the present invention, with the combination of the MRSA and drug-resistant gene triple PCR rapid amplification detection reagent, the total detection time is only 35 min, the time is saved by 55 min or more than 55 min compared with the lysozyme method, and the MRSA and drug-resistant gene nucleic acid POCT rapid detection can be achieved;

The present invention provides a 4-vinylguaiacol oxygenase mutant for synthesizing vanillin, and an engineered bacterium thereof. The 4-vinylguaiacol oxygenase mutant is an enzyme having the sequence as shown in SEQ ID NO. 1, which exhibits a double-site mutation of D27P / T216M, D27P / M351F, D27P / Y156D, M351F / Y156D, T216M / Y156D or T216M / M351F, or a single-site mutation of T216M, T216L, S280M, G307P or M351F, and can be used for catalyzing 4-vinylguaiacol to generate vanillin.

The invention relates to an Escherichia coli engineering strain for producing 2, 3-pentanediol as well as a construction method and application of the Escherichia coli engineering strain, and the method comprises the following steps: constructing recombinant plasmids pACYC-IlvA *-ThrA * BC for expressing an IlvA * gene of threonine deaminase, a ThrA gene of asparaginic acid kinase I, a Thr B gene of homoserine kinase and a ThrC gene of threonine synthetase; the method comprises the following steps: constructing a recombinant plasmid pRSF-Ilv2C-Aro10 for expressing an Ilv2C gene of acetohydroxy acid synthetase and an Aro10 gene of propiophenone dehydrogenase; and chemically converting the recombinant plasmid pACYC-IlvA *-ThrA * BC and the recombinant plasmid pRSF-Ilv2C-Aro10, and transferring the recombinant plasmid pACYC-IlvA *-ThrA * BC and the recombinant plasmid pRSF-Ilv2C-Aro10 into escherichia coli so as to obtain the escherichia coli engineering strain for producing the 2, 3-pentanediol. According to the escherichia coli engineering strain, the biosynthesis of 1.61 g / L of 2, 3-pentanediol is realized by using L-threonine and 40 g / L of glucose.

The invention discloses saccharomyces cerevisiae MBPSc21 and application thereof, and belongs to the technical field of microorganisms. On one hand, the invention provides a saccharomyces cerevisiae engineering strain MBPSc21 for expressing immune nutrient-beta-carotene, and the preservation number of the saccharomyces cerevisiae engineering strain MBPSc21 is CGMCC No.32663; on the other hand, the invention provides application of the saccharomyces cerevisiae, the strain takes soybean meal as a solid-state fermentation substrate, the crude protein content of the obtained product fermented soybean meal is remarkably increased, anti-nutritional factors such as oligosaccharide and urease are greatly reduced, more importantly, the fermented soybean meal is rich in beta-carotene, and the content of the beta-carotene is reduced. The soybean meal protein is endowed with new functional immune nutrition, so that the overall nutritional value of the soybean meal is improved, and a leading level is achieved in the field of developing functional fermented soybean meal by utilizing saccharomyces cerevisiae.

The invention provides a method for producing borneol, the method comprising culturing the transgenic microorganism in a culture medium, and the microorganism comprises or expresses borneol diphosphate synthetase and phosphatase.

The invention provides a saccharomyces cerevisiae engineering strain HYB05 capable of releasing astaxanthin in a controllable self-splitting manner. The saccharomyces cerevisiae engineering strain HYB05 is constructed by taking a basic strain for producing astaxanthin as a starting strain, introducing a coding sequence comprising cell wall lyase Lyc, a coding sequence of a signal peptide mutant F8A and a recombinant nucleic acid construct for expressing a regulatory element, and knocking out an SSD1 gene to relieve a translation inhibition effect. The cell wall lyase Lyc is found and disclosed by the inventor for the first time, and the amino acid sequence is shown as SEQ ID NO: 6; the amino acid sequence of the signal peptide mutant F8A is as shown in SEQ ID NO: 8. According to the engineering strain HYB05, by introducing the recombinant nucleic acid construct, the activation of the catalytic activity of cell wall lyase under specific conditions is realized, so that the cell wall of the saccharomyces cerevisiae is broken, and the astaxanthin synthesized by the saccharomyces cerevisiae is released. Compared with the prior art, the astaxanthin is produced through fermentation of the strain, the extraction steps are simplified, extraction of the astaxanthin in the yeast is achieved to the maximum extent, the yield of the astaxanthin is remarkably increased, and the technical effect is remarkable.

An engineered bacterium for improving tryptophan yield, a construction method therefor, and a use thereof. A strain capable of tolerating a high concentration of tryptophan was obtained by screening. Genome sequencing and protein sequence analysis of the strain revealed that some proteins in the strain had undergone point mutations, and these mutants can improve the tryptophan yield. In order to further improve the tryptophan yield, modifications are made to a protein sequence expressed by a fadR gene or a protein sequence expressed by a pepD gene in a starting bacterium. Such modifications enable the obtained engineered bacterium to have a higher tryptophan yield than the starting bacterium. In the case of large-scale production, in a 5 L fermentation tank, the tryptophan yield can reach 62.38±5.80 g / L, and the sugar-to-acid conversion rate can reach 24.1%. Compared with the original bacterium, the tryptophan yield is increased by 1.48 folds, and the sugar-to-acid conversion rate is increased by 1.26 folds. The biological material and the use thereof relate to the technical field of molecular biology and have wide practical application value.

The invention provides engineering lyase Art-15 with broad-spectrum antibacterial activity, antibacterial peptide NZ2114 and bacteriophage lyase PlySs2 are subjected to functional fusion through an optimally designed flexible linker, and an efficient bactericidal effect on Staphylococcus aureus (Staphylococcus aureus) and Streptococcus spp.) is achieved. The engineering lyase Art-15 has the advantages that the engineering lyase Art-15 has broad-spectrum antibacterial activity, the antibacterial peptide NZ2114 and the bacteriophage lyase PlySs2 are subjected to functional fusion through the optimally designed flexible linker, and the engineering lyase Art-15 has broad-spectrum antibacterial activity; the lyase Art-15 overcomes the defects of narrow host spectrum and insufficient splitting activity of the lyase through a synergistic destruction mechanism of targeting bacterial cell wall peptidoglycan. Experiments show that the minimal inhibitory concentration (MIC) of the lyase Art-15 on staphylococcus aureus in vitro is lower than 2 mu g / mL, the minimal inhibitory concentration of the lyase Art-15 on streptococcus agalactiae is lower than 1 mu g / mL, the minimal inhibitory concentration of the lyase Art-15 on streptococcus dysgalactiae is lower than 2 mu g / mL, and the sterilization rate of the lyase Art-15 can reach 97% or above within 30 minutes by 2 times of MIC. The lyase Art-15 disclosed by the invention has important application value in the fields of drug-resistant bacterium infection treatment, medical instrument disinfection and livestock breeding.

The invention provides a phaffia rhodozyma mutant strain for high yield of astaxanthin as well as a screening method and application of the phaffia rhodozyma mutant strain, belongs to the technical field of fermentation microorganism screening, and can solve the problems of low astaxanthin yield, high cost and low growth temperature of wild type phaffia rhodozyma existing in synthesis of astaxanthin from a phaffia rhodozyma strain. The invention provides a phaffia rhodozyma mutant strain capable of producing astaxanthin at high yield, the phaffia rhodozyma mutant strain is a phaffia rhodozyma AA0529 mutant strain, the phaffia rhodozyma AA0529 mutant strain is preserved in China Center for Type Culture Collection in Wuhan, Hubei on September 7, 2023, and the preservation number is CCTCC (China Center for Type Culture Collection) NO: M20231637. According to the method, the yield of the astaxanthin can be increased, and the yield of the astaxanthin is increased to 3.3 mg / g at 26 DEG C.

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 a method for biological high-density synthesis of retinaldehyde and a synthetic strain thereof. The synthetic strain is obtained by expressing ATP citrate lyase ACL, acetyl-CoA synthase ACS, choline kinase CK, inositol polyphosphate kinase IPK and vitreoscilla hemoglobin VHb in host bacteria. Wherein the host bacterium is a pichia pastoris gene modified bacterial strain PP-B2C. The retinaldehyde production performance of the recombinant strain is verified on the basis of a precursor supply pathway, an IUP pathway and a dissolved oxygen level, and the retinaldehyde production capacity of the pichia pastoris is further improved. The construction method of the recombinant pichia pastoris is simple, the synthesis of retinaldehyde can be better promoted, and the retinaldehyde can be efficiently synthesized by the engineering strain by utilizing glucose and methanol through amplification fermentation of a 5L fermentation tank, so that the recombinant pichia pastoris is beneficial to industrial production.

The invention discloses a bacteriophage lyase and application thereof, and particularly relates to a bacteriophage lyase with an amino acid sequence as shown in SEQ INNO.2. The bacteriophage lyase can inhibit the growth of escherichia coli, staphylococcus and salmonella, can be used as an antibacterial substance in splitting gram-negative bacteria and can be used for preparing drugs for resisting gram-negative bacteria. The lyase can be used independently or compounded with other substances, the use concentration of antibiotics can be reduced by combining the lyase with the antibiotics, toxic and side effects (such as renal toxicity of polymyxin) of drugs are reduced, and meanwhile, generation of bacterial drug resistance is delayed. After the lyase and the chitosan are combined for use, remarkable antibacterial activity is generated, the antibacterial effect can be achieved without pre-treatment on bacteria, and the practicability is improved.

The invention discloses a method for preparing butanediamine and spermidine from whole cells, and belongs to the field of bioengineering. According to a rational copolymerization modification strategy, a meridian interface and a latitudinal channel interface of arginine decarboxylase are subjected to rational modification respectively or simultaneously, and a series of mutants are obtained. The optimal mutant AdiA H729D / E467K / H736E, which is subjected to double-interface synergistic modification, disclosed by the invention, keeps high activity in the whole neutral-alkaline range of pH (Potential of Hydrogen) of 7.0 to 9.0. The optimal pH of the arginine decarboxylase mutant is increased, the enzyme activity stability in a neutral pH range is improved, the arginine decarboxylase mutant is more suitable for the condition requirements of industrial microbial fermentation, and a foundation is laid for efficient synthesis of butanediamine. The spermidine yield is up to 153.3 mg / L by using the mutant to catalyze the whole cell.

The invention provides low-temperature alginate lyase and application thereof, namely alginate lyase with low-temperature catalytic activity, which is separated from marine-derived cold monad bacteria, and the amino acid sequence of the alginate lyase is SEQ ID NO: 1; the amino acid sequence of the alginate lyase after the signal peptide is removed is SEQ ID NO: 3. The algin lyase Alg3889 provided by the invention has the optimal reaction temperature of about 10-16 DEG C, has good temperature stability, and still keeps more than 70% of enzyme activity after being incubated at 0-70 DEG C for 1 hour; the optimum pH value of the strain is 8.0, after the strain is incubated in a buffer system with the pH value of 3.0-10.6 for 12 h, about 90% of the highest enzyme activity can be kept, and the strain has high acid resistance and alkali resistance. The degradation product of the alginate oligosaccharide is alginate oligosaccharide with the polymerization degree of 2-6, and the alginate oligosaccharide has high application value.

The invention provides a genetically engineered bacterium for producing astaxanthin, and the genetically engineered bacterium takes Yarrowia lipolytica Po1f as an original strain and contains a GGS gene, an HMGR gene, a beta-carotene synthesis gene, a beta-carotene ketolase gene and a beta-carotene hydroxylase gene. The invention further provides the Yarrowia lipolytica MC247, and the preservation number of the Yarrowia lipolytica MC247 is CGMCC NO.29699. According to the Yarrowia lipolytica MC247, the high-yield astaxanthin is obtained, a new technical scheme is provided for efficient and green manufacturing of the astaxanthin, and industrial production of the astaxanthin is facilitated.

Herbicide-resistant rice plants, isolated polynucleotides that encode herbicide resistant and wild-type acetohydroxy-acid synthase large subunit 1 (AHASL1) polypeptides, and the amino acid sequences of these polypeptides, are described. Expression cassettes and transformation vectors comprising the polynucleotides of the invention, as well as plants and host cells transformed with the polynucleotides, are described. Methods of using the polynucleotides to enhance the resistance of plants to imidazolinone herbicides, and methods for controlling weeds in the vicinity of herbicide-resistant plants are also described.

Described herein are biocatalytic methods of producing terpene degradation products useful as starting material for the production of perfumery ingredients, such as, for example, ambrox. In particular novel terpene degrading polypeptides (enal-cleaving polypeptides) and novel peptides converting terpenes compounds to oxygenated derivatives (oxygenases) and mutants and variants derived therefrom are described which may be applied in novel types of fully enzymatic multistep degradation pathways allowing the controlled, stepwise conversion and degradation of linear or cyclic terpene substrates. Said novel biosynthetic strategies allow the fully biochemical synthesis of valuable terpene-derived compounds, like for example manooloxy or gamma ambrol. Also described herein are recombinant host organisms carrying the required set of genetic information for the functional expression of the set of enzymes necessary for catalyzing the combination of enzymatic conversion and degradation steps.

The invention belongs to the field of metabolic engineering, and discloses a genetically engineered bacterium for producing myo-inositol as well as a construction method and application of the genetically engineered bacterium. The genetically engineered bacterium takes escherichia coli as an original strain, and is obtained by performing the following gene editing on a genome of the escherichia coli: non-expressed lactose operon repressor protein, glucose phosphate dehydrogenase, acetokinase, glucose phosphate isomerase and glycerol repressor protein; and overexpressing inositol-1-phosphate synthase, inositol monophosphate, glucokinase, glucose permease and glycerol kinase. Compared with the prior art, the genetically engineered bacterium disclosed by the invention has remarkable advantages in the aspects of key enzyme expression rate, metabolism specificity, fermentation period and the like, and an efficient, stable, economical and feasible solution is provided for industrial production of myo-inositol.

The invention discloses a Pickering IL emulsion microreactor constructed on the basis of temperature-sensitive double-enzyme Janus spheres and application of the Pickering IL emulsion microreactor to enhanced catalysis of conversion of CO2 into formic acid. A Janus emulsion template method is combined with a photocuring technology to synthesize a nano-scale dual-functional temperature-sensitive Janus sphere, one half of the nano-scale dual-functional temperature-sensitive Janus sphere is a hydrophilic hydrocarbon polymer, the other half of the nano-scale dual-functional temperature-sensitive Janus sphere is a hydrophobic fluorocarbon polymer, and rapid inversion of a Pickering emulsion can be realized by utilizing temperature sensitivity. The method comprises the following steps: preparing a Janus particle hydrophilic / hydrophobic hemisphere, selectively immobilizing formate dehydrogenase FDH and carbonic anhydrase CA on the Janus particle hydrophilic / hydrophobic hemisphere to obtain a temperature-sensitive FDH-Janus-CA composite sphere, introducing a fluorine-functionalized ionic liquid to regulate and control the internal microenvironment of a microreactor, and constructing a Pickering IL emulsion microreactor. The CO2 absorption capacity is increased by 47 times compared with that of a pure water system, the formic acid yield reaches 27.5 mM and is higher than the maximum value 22.8 mM of current enzyme electrocatalysis, the storage and catalysis stability is good, and an efficient and sustainable scheme is provided for CO2 recycling.

The invention belongs to the technical field of biology, and particularly relates to a construction method and application of a tobacco plant for expressing rare ginsenoside. According to the method, dammarendiol synthase DDS, cytochrome P450 CYP716A47 and glycosyl transferase UGTPg1 / UGTPg45 are added into tobacco, heterologous synthesis of rare ginsenosides CK and Rh2 is achieved, dependence on traditional ginseng cultivation extraction is avoided, and cost is remarkably reduced and efficiency is remarkably improved. The characteristics of short growth cycle and easy culture of tobacco are utilized to overcome the problems of low efficiency and low yield of traditional synthesis; positive transgenic tobacco leaves are dedifferentiated to form callus cells, so that materials are provided for large-scale fermentation, and an industrial production foundation is laid.

The present invention relates to, generally, modified Streptomyces cells which produce alligamycin A with higher yields.

The invention belongs to the technical field of natural product extraction and enzyme engineering, and particularly relates to a method for extracting red pomelo polyphenol based on an enzyme method. The method aims at solving the problems of high cost and the like caused by easy inactivation and difficult recovery of enzymes in most red pomelo extraction processes. Waste such as pericarp and pomelo seeds generated by red pomelo processing are used as carbon sources, a biochar material with a hierarchical porous structure is prepared through programmed heating carbonization activation, pectinase, cellulase and naringinase are immobilized by adopting a spatial zoning sequential immobilization strategy and then react with the red pomelo, and the biochar material with the hierarchical porous structure is prepared. The immobilized enzyme is rapidly recovered through magnetic separation to realize repeated utilization of the enzyme, and finally red pomelo polyphenol powder is obtained through multi-step treatment, so that recycling is realized, and the yield of polyphenol extracted by an enzyme method is improved.

The invention discloses a bergamot alkene synthase mutant and application thereof, and belongs to the technical field of enzyme engineering and bioengineering. According to the method, bergamotene synthase SanSyn which is obtained from plant clausena lansium and can catalyze FPP to generate alpha-trans-bergamotene is taken as an object, semi-rational design and site-specific mutagenesis are carried out on the bergamotene synthase SanSyn, a series of obtained SanSyn mutants are remarkably improved in the aspect of FPP substrate conversion efficiency, and the optimal bergamotene synthase mutant SanSynI273C-L514I has the advantages that the optimal bergamotene synthase mutant SanSynI273C-L514I can be used for catalyzing the FPP to generate alpha-trans-bergamotene; the unit cell productivity of the alpha-trans-bergamotene generated by catalyzing FPP is 6.9 times of that of a wild type, and the yield is 6.8 times of that of the wild type and reaches 43.58 mg / L. According to the bergamot alkene synthase mutant, the performance of alpha-trans-bergamot alkene synthesized by microorganisms is enhanced, and an effective technical route is provided for green production of industrial high-purity bergamot alkene.

The embodiment of the invention provides a tyrosine ammonia lyase mutant and application thereof in synthesis of p-coumaric acid. According to the embodiment of the invention, an amino acid sequence as shown in SEQ ID NO.2 is taken as a template, a key active site of the template is subjected to mutation transformation, a mutant library is constructed, and a series of mutants with remarkably improved catalytic activity and / or conversion rate are obtained through screening; the tyrosine ammonia lyase mutant provided by the embodiment of the invention can selectively catalyze L-tyrosine to generate p-coumaric acid, has excellent yield and / or higher yield, and has outstanding industrial application prospects in efficient and green synthesis of p-coumaric acid.