201 results about "Acyltransferase" patented technology

The invention provides a Yarrowia lipolytica engineering strain for producing cocoa butter equivalent and a construction method and application thereof, and relates to the technical field of biology, the construction method comprises the following steps: inserting an anti-translation PFAD2 promoter into a Yarrowia lipolytica chassis strain to adjust the composition of target fatty acid, so that the contents of palmitic acid and stearic acid are respectively increased to 27.3% and 19.8%; the contents of palmitoleic acid and oleic acid are reduced to 4.5% and 36.8%; the twelve-site fatty acid desaturase gene FAD2 is knocked out, and linoleic acid is completely eliminated; a plant-derived diacylglycerol acyltransferase gene DGAT or phospholipid is introduced; the diacylglycerol acyltransferase gene PDAT is used for synthesizing cocoa butter equivalent (CBE). The yield of cocoa butter equivalent of the constructed yarrowia lipolytica engineering strain finally reaches 1.00 + / -0.09 g / L and accounts for 30% of the total fat content. The technical problem of low cocoa butter equivalent production efficiency of engineering modified yeast in the prior art is solved.

The invention relates to the technical field of microbial metabolism engineering, in particular to recombinant escherichia coli for efficiently producing O-acetyl-L-homoserine as well as a construction method and application of the recombinant escherichia coli. The invention relates to recombinant escherichia coli for efficiently producing O-acetyl-L-homoserine, which is characterized in that a transport protein encoding gene rhtA is knocked out in a chassis strain Escherichia coli W3110, a homoserine O-acetyltransferase encoding gene metXcm from marine cyclobacter is over-expressed, an acetyl coenzyme A synthetase encoding gene acs is over-expressed, pantothenic acid kinase coXpp from pseudomonas putida is over-expressed, and the recombinant escherichia coli can be used for efficiently producing O-acetyl-L-homoserine. And knocking out the malate synthetase coding gene aceB to obtain the recombinant escherichia coli for efficiently producing the O-acetyl-L-homoserine. The strain provided by the invention does not carry plasmids, does not need to additionally add antibiotics and inducers, and is lower in production cost and more stable. And the yield of the O-acetyl-L-homoserine of the plasmid-free strain is increased to 16.85 g / L from 0 g / L. Therefore, the recombinant escherichia coli provided by the invention has important industrial application value.

The present invention provides an α-amino acid ester acyltransferase mutant and its application. The α-amino acid ester acyltransferase mutant includes: (a) a protein having the amino acid sequence shown in SEQ ID NO: 1; (b) a protein having an amino acid mutation at at least one of the following sites in the amino acid sequence in (a): P158, K80, T208, A302, A175, N85, N605, K346, S225, I34, or S348, and having the function of an α-amino acid ester acyltransferase; or (c) a protein having more than 80% homology with the amino acid sequence defined in (a) or (b) and having the function of an α-amino acid ester acyltransferase. The α-amino acid ester acyltransferase mutant of the present invention has the advantages of a broad substrate spectrum and high catalytic activity, can be well used in the synthesis of oligopeptides, has low cost and high yield, and realizes true green chemistry.

The invention provides a euphorbia lathyris acyltransferase gene ElBAHD16 and application of a coding protein thereof. The gene is obtained by cloning from euphorbia lathyris for the first time. The invention also provides a recombinant vector and an engineering bacterium containing the gene. Cinnamoyl coenzyme A, benzoyl coenzyme A or acetyl coenzyme A can be used as an acyl donor, and 7-hydroxy euphorbia lathyris diterpenoid alcohol is used as a substrate to generate a plurality of types of euphorbia lathyris diterpenoid esters through biological catalysis. The content of diterpenoid ester substances in plants such as euphorbia lathyris can be increased through a genetic engineering technology, and the method has a good application prospect.

The invention relates to the technical field of medicine analysis, in particular to a method for analyzing mutagenic impurities in a small molecule fungal inositol acyltransferase 1 inhibitor raw material medicine, the analysis method can be used for trace detection of potential mutagenic impurities I and II in the raw material medicine, the limit of quantitation concentration of the impurity I is not higher than 0.53 ng / ml, and the limit of quantitation concentration of the impurity II is not higher than 0.53 ng / ml. The detection limit concentration of the impurity I is not higher than 0.27 ng / ml; the quantitation limit concentration of the impurity II is not higher than 0.52 ng / ml, and the detection limit concentration of the impurity II is not higher than 0.26 ng / ml. The analysis method suitable for the mutagenic impurities in the small-molecule fungal inositol acyltransferase 1 inhibitor bulk drug is high in sensitivity and high in specificity, and the blank solvent does not interfere with detection of the impurities.

The present invention discloses a recombinant Escherichia coli producing α-amino acid ester acyltransferase, a construction method and an application thereof, and belongs to the field of biotechnology. The recombinant Escherichia coli is deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration Committee, with a deposit address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, a deposit date of March 22, 2024, and a deposit number of CGMCC No. 30105. The recombinant Escherichia coli constructed by the present invention can produce α-amino acid ester acyltransferase, and the enzyme is used to catalyze the synthesis of glutathione, with a conversion rate of up to 82.59%, which is of great significance to the industrial development of biosynthesis of glutathione.

The invention discloses acyltransferase, a mutant and application in preparation of acetyl harpagide, the acyltransferase ancestor sequence capable of biologically catalyzing the acetylation of harpagide is firstly screened, then the acyltransferase is subjected to mutation modification, the enzyme activity is further improved, the harpagide is used as a substrate, and the acyltransferase and the mutant are synthesized into the acetyl harpagide. The ancestor acyltransferase and the mutant are used for acylation reaction, so that the application of the acyltransferase in the fields of medicines and the like is promoted, and certain help is provided for the aspect of drug acylation modification.

The invention discloses a method for biologically converting isorhamnetin, which comprises the following steps: adding a microorganism for producing glucosidase, glycosyl transferase and / or succinyl transferase, or producing glucosidase, glycosyl transferase and / or succinyl transferase into an isorhamnetin substrate, adding a glycosyl donor for synchronous conversion, and separating and purifying, thereby obtaining the isorhamnetin. And the isorhamnetin-3-O-glucoside, the isorhamnetin-3-O-beta-(6 ''-O-succinyl)-D-glucoside and / or the isorhamnetin-3, 7-O-diglucoside are obtained after the reaction is finished. The method is green, safe and high in conversion efficiency, the product has higher water solubility, the bioavailability and additional value of the flavone compound can be improved, and high-valued utilization of natural plant product raw materials is achieved.

The invention discloses a 7-aminocephalosporanic acid producing strain as well as a preparation method and application thereof. The invention provides a method for preparing 7-aminocephalosporanic acid. The method comprises the following steps: carrying out combined plasmid construction on promoters from different sources and a plurality of cephalosporin C acyltransferases; a cephalosporin C acyltransferase gene is integrated into a cephalosporium acremonium strain genome through a protoplast transformation method to obtain recombinant bacteria, and the recombinant bacteria are fermented to obtain a 7-aminocephalosporanic acid product. According to the invention, CPC high-productivity cephalosporium acremonium strain is taken as a starting strain, and cephalosporin C acyltransferase is introduced into the starting strain in an exogenous manner by means of genetic engineering, synthetic biology and the like, so that direct production of 7-aminocephalosporanic acid from the engineering strain can be realized. The method has the advantages of simple steps, no chemical pollution in the production process, and huge industrialization prospect.

The invention provides an immobilized enzyme as well as a preparation method and application thereof. The immobilized enzyme comprises an enzyme and a carrier for loading the enzyme, wherein the enzyme is alpha-amino acid fatty acyl transferase or polypeptide ligase; the carrier is a polymethacrylate matrix carrier; the polymethacrylate matrix carrier comprises an amino carrier or an epoxy carrier; wherein the amino carrier is an amino carrier modified by a long-chain polymer; the epoxy carrier is an epoxy carrier modified by a long-chain polymer; wherein the long-chain polymer comprises polyethylene glycol (Mw) of 2-8 KDa, polyvinyl alcohol (Mw) of 1.7-31 KDa, polyethyleneimine (Mw) of 1.8-70 KDa, polyacrylamide (Mw) of 50-300 KDa or polylysine (Mw) of 1.6-41 KDa. The immobilized enzyme disclosed by the invention is good in intermolecular dispersity and relatively large in spacing, can efficiently catalyze a substrate to synthesize oligopeptide or polypeptide, and has relatively high substrate conversion rate.

The invention discloses an escherichia coli engineering strain with high benzyl acetate yield and application thereof. The escherichia coli strain efficiently expresses a brand new biosynthetic pathway related gene of benzyl acetate; comprising a phenylalanine lyase gene, a cinnamic acid coenzyme A ligase gene, a 3-hydroxyacyl coenzyme A dehydrogenase gene, a 3-oxoacyl coenzyme A ketohydrolase gene, an enoyl coenzyme A hydratase gene, a carboxylic acid reductase gene, a phosphoric acid pantetheinyl transferase gene and an acyltransferase gene. The obtained escherichia coli engineering strain can utilize a sustainable and cheap raw material glucose as a carbon source to realize efficient synthesis of benzyl acetate. The yield of benzyl acetate synthesized by the escherichia coli benzyl acetate engineering bacteria is the highest yield of benzyl acetate synthesized by a microbial fermentation method reported at home and abroad from beginning. The benzyl acetate produced by the engineering strain has the advantages of reliable quality, economic product, green production and the like, and has important economic value and social benefit.

The invention discloses a mutant of acylase and application of the mutant in synthesis of capryloyl glycine. The amino acid sequence of the acylase mutant is obtained by mutating at least one of E155D, Y160G and A330D from a sequence shown as SEQ ID NO: 2, the catalytic activity of the acylase mutant is remarkably improved compared with that of wild type acylase, and the thermal stability of the acylase mutant is excellent. The mutant of the acylase provided by the invention provides a solid foundation for industrial production of capryloyl glycine prepared by enzyme catalysis.

The invention relates to construction and dyeing application of recombinant Escherichia coli for synthesizing indole acetate, which is characterized in that recombinant Escherichia coli MG1655 delta pheA delta trpR delta tyrA delta yddG delta arsB: tac 70-MaFMO delta ldhA: tac 70-MaFMO delta yghWX: tac 70-tnaA delta poxB: tac 70-cav1 is taken as an initial strain, then endogenous acetylesterase is knocked out, and the recombinant Escherichia coli for synthesizing indole acetate is obtained by the steps of: preparing the recombinant Escherichia coli for synthesizing indole acetate. An endogenous tryptophan hydrolase gene tnaA and a flavin-containing monooxygenase gene maFMO are overexpressed, and meanwhile, an acetyltransferase gene is overexpressed, so that the tryptophan-containing flavin-containing monooxygenase gene is obtained; the yield of the indole acetate reaches up to 195.3 mg / L. When the indole acetate is used for dyeing, a reducing agent and an enzyme solution are not used, so that the dyeing cost is greatly reduced.

The invention relates to the field of biosynthesis high polymer materials, in particular to a biosynthesis method and application of omega-hydroxy fatty acid polyester (omega-PHFA) based on a whole-cell catalytic system. The method comprises the following steps: constructing a recombinant microbial whole-cell catalysis system containing P450 monooxygenase, CoA ligase and acyltransferase, and synthesizing omega-PHFA with a C12-C18 long carbon chain skeleton by taking renewable fatty acid as a substrate. Compared with a traditional chemical / enzymatic synthesis method, the technology is greener, and the performance defects of existing biosynthetic PHA materials are overcome. The invention provides related genetically engineered bacteria and a method for producing omega-PHFA by using the genetically engineered bacteria, and the genetically engineered bacteria have a wide application prospect.

Compounds and compositions comprising compounds that modulate histone acyltransferase (HAT). Methods of treating neurodegenerative disorders, conditions associated with amyloid-beta peptide deposit accumulation, Tau protein levels, and / or alpha-synuclein accumulation, and cancer by administering to a subject a compound that modulates HAT.

The invention discloses a recombinant bacterium capable of increasing the yield of alpha-bisabolol as well as a preparation method and application of the recombinant bacterium, and belongs to the technical field of microorganisms. The invention aims to improve the yield of alpha-bisabolol and enhance the tolerance of a host to an organic solvent. The invention provides a recombinant bacterium for improving the yield of alpha-bisabolol. Escherichia coli is used as a starting strain; the method comprises the following steps of: overexpressing an acetyl CoA acyltransferase / HMG-CoA reductase mvaE gene, an HMG-CoA synthetase mvaS gene, a 2-methyl citrate dehydratase prpD gene, a mevalonate kinase ERG12 gene, a mevalonate 5-phosphate kinase ERG8 gene, a mevalonate 5-diphosphate decarboxylase ERG19 gene and an isopentenyl diphosphate isomerase idi gene, so as to obtain a recombinant vector; the gene is obtained from an alpha-bisabolol synthase gene of artichoke, a farnesyl diphosphate synthase ispA gene and an alpha-bisabolol synthase CcBOS gene of artichoke. The industrial process of synthesizing alpha-bisabolol by a biological method is promoted.

The application provides a recombinant Yarrowia lipolytica capable of producing palmitoleic acid at a high yield, a construction method and application thereof, and belongs to the technical field of bioengineering. Yarrowia lipolytica The recombinant Yarrowia lipolytica is obtained by knocking out peroxisome biogenesis factor 10 and triacylglycerol lipase 4 in the genome of Yarrowia lipolytica and inserting an acetyl-CoA carboxylase, a Δ9 desaturase, an acetyl-CoA diacylglycerol acyltransferase, a glycerol-3-phosphate acyltransferase, a lysophosphatidylcholine acyltransferase and a phosphatidylcholine:diacylglycerol acyltransferase gene expression cassette. Experiment proves that the recombinant Yarrowia lipolytica can efficiently ferment and produce palmitoleic acid, and realizes efficient synthesis of the natural product palmitoleic acid from plants in Yarrowia lipolytica.

The present application relates to a kind of whole cell catalytic synthesis N-acetyl-trans-4-hydroxyproline method, belong to biological catalysis technical field.The present application is constructed by mutating acyltransferase and heterologous expression gene engineering bacteria, successfully realized using whole cell catalytic synthesis N-acetyl-trans-4-hydroxyproline, avoids the problem of enzyme instability, reduces the separation cost of enzyme and product.On this basis, the present application improves the yield of N-acetyl-trans-4-hydroxyproline by optimizing catalytic time, cell addition amount, the type and addition amount of additive, and the yield reaches 239mg / L.The present application is a kind of N-acetyl-trans-4-hydroxyproline preparation method with lower cost, lower equipment demand, more suitable for industrial production.

The present invention relates to tomato plants having improved insect resistance, more particularly whitefly resistance or mite resistance, wherein the plants comprise an ASAT3 gene encoding an acetyl-CoA-dependent acyltransferase and an AP2e gene encoding an APETALA2 ethylene response transcription factor. The invention also relates to a method for providing tomato plants with improved insect resistance, and the use of the ASAT3 gene in combination with the AP2e gene for providing insect resistant tomato plants.

The invention relates to the field of genetic engineering and fermentation engineering, and discloses a genetically engineered bacterium for producing melatonin and application of the genetically engineered bacterium in fermentation production of melatonin. According to the invention, genes of mtrA, PTPS, SPR, PCD and DHPR of related enzymes synthesized by coding 2-amino-6-(1, 2-dihydroxypropyl)-5, 6, 7, 8-tetrahydro-4 (1H)-pterin diketone and genes of L-2-amino-3 (beta-indole) propionic acid monooxygenase, oxymethyltransferase, N-acetyltransferase and decarboxylase are co-expressed on the genetically engineered bacterium, so that the gene engineering bacterium is obtained. The strain can be used for fermentation production of melatonin, and experiments show that the yield of the strain can reach 6480 mg / L.

The present invention relates to the use of carnitine palmitoyltransferase 1 inhibitors in the treatment of heart failure with preserved ejection fraction. The use in particular is the use of the carnitine palmitoyltransferase (CPT1) inhibitor Etomoxir for the prevention and / or treatment of heart failure with preserved ejection fraction by oral administration.

The present invention is concerned with a Diacylglycerol-acyltransferase 1 (DGAT1) inhibitor for use in the treatment of cancer in female patients.

The invention relates to an immobilization method and application of an alpha-amino acid ester acyltransferase immobilized enzyme, and belongs to the technical field of biological enzymes. In order to solve the problems of poor immobilization and difficulty in removing miscellaneous enzymes in the prior art, the invention provides an immobilization method of an alpha-amino acid ester acyltransferase immobilized enzyme, which comprises the following steps: under the catalytic action of a TG enzyme, carrying out enzyme protein cross-linking immobilization reaction on alpha-amino acid ester acyltransferase containing a plurality of glutamine residues and gelatin to obtain the alpha-amino acid ester acyltransferase immobilized enzyme. The immobilized alpha-amino acid ester acyltransferase immobilized enzyme is obtained. According to the present invention, the TG enzyme can well catalyze the glutamine residue and the lysine residue of the target protein to form the cross-linking reaction, and other miscellaneous enzymes are distinguished so as to avoid the reaction of the miscellaneous enzymes and gelatin, such that the good selectivity and the good specificity are provided, and the purification purpose can be easily achieved.

The invention discloses yarrowia lipolytica engineering bacteria for producing esterified astaxanthin as well as a construction method and application of the yarrowia lipolytica engineering bacteria. According to the invention, acyltransferase ScDGAT2-1 from fission vibrio is integrated in a genome of a yarrowia lipolytica engineering strain DN20, diacylglycerol acyltransferase DGA1 and phosphatidic acid phosphatase PAH1 are knocked out, and the corn oil lipoprotein Oleosin and the acyltransferase ScDGAT2-1 from fission vibrio are subjected to fusion expression, so that the yarrowia lipolytica engineering strain DN20 is obtained. According to the present invention, the Yarrowia lipolytica engineering bacterium capable of producing esterified astaxanthin at a high yield is constructed by using SBT06-005 as a raw material and integrating beta-carotene hydroxylase CrtZ, endogenous truncated 3-hydroxy3-methylglutaryl coenzyme A reductase tHMG1, phytoene synthase CarRP and beta-isopropyl malate dehydrogenase LEU2, and the SBT06-005 can produce 166.6 mg / L esterified astaxanthin and 1097.0 mg / L total astaxanthin in a 5L fermentation tank.

The application discloses an engineering bacterium for producing N-acetyl-trans-4-hydroxyproline and application thereof and belongs to the technical field of biotechnology.The application realizes fermentation synthesis of N-acetyl-trans-4-hydroxyproline by taking proline as a substrate through mutation of acyltransferase and introduction of exogenous proline-4-hydroxylase.The application has the advantages of low production cost, short fermentation period and high strain stability and has high economic benefits.The recombinant strain constructed by the application can convert 0.5M of proline into 19.64mg / L of N-acetyl-trans-4-hydroxyproline.

The present application belongs to the technical field of quorum quenching enzyme preparation, and particularly relates to a multi-target quorum quenching enzyme preparation and a preparation method and application thereof, such as being mainly used for inhibiting the virulence factors of Pseudomonas aeruginosa, and including acyl homoserine lactone acyltransferase AiiO protein and 3-hydroxy-4-oxoquinolone 2,4-dioxygenase AqdC protein, and the mass ratio of the two is 20:(1-25); such as being mainly used for inhibiting the virulence factors and the synthesis of biological membranes of Pseudomonas aeruginosa, and further including deferiprone, and the mass ratio of AiiO, AqdC and deferiprone is 20:(1-25):(0.695-2.78), the present application can target the Las, Rhl and PQS of Pseudomonas aeruginosa, the yield and stability of the prepared AqdC protein are both high, the two types of enzymes are used in combination after the regulation amount, and the virulence factors regulated by different quorum sensing pathways in Pseudomonas aeruginosa are effectively inhibited; the regulation amount of deferiprone is used in combination with the two types of quorum quenching enzymes, and the virulence and biological membranes of Pseudomonas aeruginosa can be effectively inhibited in a comprehensive manner.

The invention belongs to the technical field of biology, and relates to a drug target for improving sensitivity of tumor cells to immunotherapy and application of the drug target. On one hand, the invention discloses application of sterol O-acyltransferase 1 and / or 2 as a target spot in preparation of drugs for inhibiting tumor cell immune escape, a new direction is provided for tumor immunotherapy drugs, and a new platform is provided for screening of the tumor immunotherapy drugs. On the other hand, the invention provides a method for predicting the postoperative recurrence rate and / or lifetime of the tumor, and the method has a wide application prospect in the field of tumor treatment.

The invention relates to the technical field of plant growth regulation, in particular to application of berberine based on ACAA targets in resisting plant viruses and promoting plant growth. The invention reveals that berberine plays a role by specifically targeting acetyl coenzyme A acyltransferase and up-regulating the expression of the acetyl coenzyme A acyltransferase for the first time. The clear compound-target-pathway mechanism overcomes the defect that the existing plant source product mechanism is unknown. As the target spot is clear, the application dosage can be accurately controlled, so that the stability and predictability of field antiviral and growth promoting effects are ensured, and a foundation is laid for large-scale agricultural application. An ACAA target is activated by berberine, and an antiviral defense mechanism and an energy metabolism growth pathway of a plant can be synchronously started, so that the plant growth is effectively promoted while virus diseases are prevented and treated. The disease resistance-growth negative correlation problem existing in agricultural production for a long time is solved, and the dual requirements of modern agriculture for precise prevention and control and yield increase and quality improvement are met.

The invention belongs to the technical field of agricultural biology, and relates to application of glycerol acyltransferase with an amino acid sequence as shown in SEQ ID NO: 1 to improvement of salt tolerance of plants, and the glycerol acyltransferase is of great significance to creation of salt-tolerant plant germplasm resources and cultivation of salt-tolerant rice varieties.

The invention relates to the technical field of biology, and discloses a genetically engineered bacterium for high yield of 1, 4-butanediol and application of the genetically engineered bacterium. Escherichia coli is used as a chassis bacterium, a gene ldhA for coding lactic dehydrogenase, a gene adhE for coding ethanol dehydrogenase, a gene pflB for coding formate lyase, and a gene gabD and a gene yneI for coding succinate semialdehyde dehydrogenase are knocked out from a chassis bacterium genome, expression of a succinate semialdehyde dehydrogenase gene sucD and a 4-hydroxybutyrate dehydrogenase gene 4hbD is strengthened, a pdhR gene is knocked out, and a recombinant plasmid is obtained. The expression of the gene gltAR164L is enhanced, the gene mdh, the gene arcA, the gene poxB, the gene sdhAB and the gene ptsG are knocked out, and the expression of the glf gene and the ppc gene is enhanced; the method comprises the following steps of: transferring plasmids carrying a gene ald for coding aldehyde dehydrogenase, a gene cat2 for coding acyltransferase and a gene yqhD for coding alcohol dehydrogenase into a chassis bacterium, and carrying out metabolic transformation to obtain a genetically engineered bacterium which carries exogenous plasmids and has the capabilities of high yield and extracellular accumulation of 1, 4-BDO, so that the yield of 1, 4-BDO can be increased to 4.71 g / L.