20 results about "Coenzyme A Ligases" patented technology

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 relates to the field of bioengineering, in particular to a fusion enzyme mutant and application thereof in fluororesveratrol synthesis. The invention provides a mutant composition. The mutant composition comprises a mutant of p-coumaroyl-CoA ligase and a mutant of stilbene synthase, the mutant of the p-coumaroyl-coenzyme A ligase is characterized in that on the basis of wild type p-coumaroyl-coenzyme A ligase, the mutant has mutation at the 113th site and / or the 537th site; the mutant of the stilbene synthase comprises mutation at the 170th site on the basis of wild type stilbene synthase. According to the invention, the fusion enzyme is subjected to directed evolution by applying a semi-rational CAST / ISM strategy and an irrational error-prone PCR strategy for the first time, a fusion enzyme mutant At4CL (F537G / F113G)-VvSTS (V170A) with improved catalytic activity is obtained, and finally, a series of fluorine-containing resveratrol is efficiently synthesized.

The invention provides a genetically engineered bacterium, a preparation method of the genetically engineered bacterium and application of the genetically engineered bacterium in synthesis of resveratrol. The genetically engineered bacterium contains a p-coumaroyl coenzyme A ligase gene At4CL and a stilbene synthase gene PcSTS, wherein the p-coumaroyl coenzyme A ligase gene At4CL is derived from arabidopsis thaliana, and the stilbene synthase gene PcSTS is derived from Polygonum cuspidatum, and the p-coumaroyl coenzyme A ligase gene At4CL is derived from Polygonum cuspidatum. Wherein the genetically engineered bacteria are selected from escherichia coli. The method for catalytically synthesizing resveratrol by using the genetically engineered bacterium is short in fermentation period, high in yield and high in production intensity, has a relatively good application prospect, and lays a foundation for industrial production of resveratrol by using an escherichia coli microbial fermentation method.

The invention discloses recombinant saccharomyces cerevisiae, a 11-seed oil fermentation product as well as a preparation method and application of the 11-seed oil fermentation product, and belongs to the technical field of synthetic biology and fermentation product preparation. According to the recombinant saccharomyces cerevisiae disclosed by the invention, a caffeic acid coenzyme A ligase 4CL coding gene, a chalcone isomerase CHI coding gene, a flavanone-3-hydroxylase F3H coding gene and a chalcone synthase CHS mutant coding gene are over-expressed, and the chalcone synthase CHS mutant is preferably Y69H / H71Y / Q161K. The preparation method of the 11-seed oil fermentation product comprises the following steps: fermenting a recombinant saccharomyces cerevisiae fermentation solution in the presence of a sophora flower bud extract and caffeic acid to obtain a first fermentation solution; fermenting the lactic acid bacteria fermentation liquor in the presence of 11 seed oil to obtain second fermentation liquor; and mixing the first fermentation liquid and the second fermentation liquid, continuing fermentation, and respectively collecting an oil phase and a water phase after fermentation is finished, so as to obtain a fermentation product. The product provided by the invention has multivitamin antioxidant activity.

The invention belongs to the technical field of biology, and discloses a saccharomyces cerevisiae engineering strain for producing naringenin, a construction method and application, the saccharomyces cerevisiae engineering strain contains tyrosine ammonia lyase gene TAL, 4-cumaric acid: coenzyme A ligase gene 4CL, chalcone synthase gene CHS, chalcone isomerase gene CHI, oligopeptide gene RIADn-RGGm and oligopeptide gene RIDD; the ARO10 gene, the FDC1 gene, the PAD1 gene and the PDC5 gene are knocked out. The engineering strain disclosed by the invention can be used for synthesizing naringenin by taking L-tyrosine as a substrate, and the engineering strain is stable in hereditary character and excellent in naringenin yield which can reach 27.45 mg / L; and the fermentation production of the product can be carried out through relatively simple method conditions, so that the method has a very wide application prospect.

A method for producing γ-Glu-Val-Gly is described, wherein the method includes the steps of cultivating a γ-Glu-Val-Gly-producing bacterium belonging to the family Enterobacteriaceae in a culture medium so that the γ-Glu-Val-Gly accumulates in the culture medium or the cells of the bacterium, or both, and collecting the γ-Glu-Val-Gly from the culture medium or the cells of the bacterium, or both. The bacterium has been modified to overexpress a gene encoding a protein having L-threonine 3-dehydrogenase activity and a gene encoding a protein having 2-amino-3-oxobutanoate coenzyme A ligase activity.

The present invention relates to a microorganism containing at least one heterologous gene encoding L-arginine:glycine amidinotransferase (AGAT), at least one heterologous L-threonine 3-dehydrogenase gene (tdh), and at least one heterologous glycine C-acetyltransferase (2-amino-3-oxobutanoate coenzyme A ligase) gene (kbl), as well as a method for the fermentative production of guanidinoacetic acid (GAA) using such a microorganism. The present invention also relates to a method for the fermentative production of creatine.

The present application relates to the field of genetic engineering and bioengineering, and particularly relates to a genetically engineered surfactin-producing bacterium and application thereof.The genome of the genetically engineered bacterium comprises the following: an expression frame of T7 RNA polymerase; an expression frame of a surfactin synthesis gene cluster srfAABCD; and an expression frame of a long-chain fatty acid-coenzyme A ligase gene lcfA; the expression frame of the surfactin synthesis gene cluster srfAABCD and the expression frame of the long-chain fatty acid-coenzyme A ligase gene lcfA further comprise a T7 promoter.The present application realizes efficient synthesis of surfactin by Bacillus subtilis, and the final shake flask yield reaches more than 19.3 g / L, which has great industrial application potential.

This invention belongs to the field of forage crop cultivation, specifically relating to a high-yield forage crop cultivation method for herbivorous animal feed. This invention utilizes both inner and outer seed-encapsulated liquids to provide forage seeds with a sufficient nutritional environment, reducing abiotic stress during growth. Trichoderma and other microorganisms in the inner seed-encapsulated liquid reproduce and absorb nutrients from the outer seed-encapsulated liquid and the external soil environment, then supply their metabolites to the forage seeds. The improved water and fertilizer system of this invention enhances soil conditions. The compound preparation of this invention inhibits the key enzyme in lignin synthesis, 4-coumaric acid-coenzyme A ligase. Applying the compound preparation after harvesting allows for precise dosage control, preventing lodging and reducing lignin synthesis. Forage obtained using this method has lower lignin content, better palatability for animals, higher yield, and better quality, thus reducing input costs in the livestock industry.

This invention belongs to the field of genetic engineering, and specifically relates to a 4-coumaric acid-coenzyme A ligase gene Gl4CL from *Gynostemma pentaphyllum*, its encoded product, and its applications. This invention is the first to clone and prepare the 4-coumaric acid-coenzyme A ligase gene Gl4CL from the medicinal plant *Gynostemma pentaphyllum*. The nucleotide sequence of this gene is shown in SEQ ID NO.1, and the amino acid sequence of the product encoded by this nucleotide sequence is shown in SEQ ID NO.2. The 4-coumaric acid-coenzyme A ligase gene Gl4CL is a key regulatory gene in the flavonoid biosynthesis pathway of *Gynostemma pentaphyllum*. This enzyme can be used in the preparation of p-coumaryl-CoA using p-coumaric acid as a substrate, further promoting the biosynthesis of flavonoid compounds. This invention can provide an effective method to meet the huge market demand for flavonoid compounds.

The present invention provides a yeast genetic engineering bacterium for producing resveratrol and a construction method and application thereof. The engineering bacterium overexpresses resveratrol synthase on the chromosome of Saccharomyces cerevisiae BY4742. STS , p-Coumaryl-CoA ligase 4CL , phenylalanine / tyrosine ammonia lyase TAL , cinnamate-4-hydroxylase C4H , cytochrome P450 enzyme reductase CPR1 , DAHP synthase mutant ARO4 , chorismate mutase mutant ARO7 Chorismate synthase ARO2 shikimate kinase AroL , acetyl-CoA synthetase ACS and malonyl-CoA synthetase ACC1 , and knock out bypass genes CIT2, LPP1, DPP1, and PDC6 at the same time, to achieve biosynthesis of resveratrol using glucose as a substrate. The present invention utilizes phenylalanine / tyrosine ammonia lyase to react with phenylalanine and tyrosine as substrates to generate key precursors for resveratrol synthesis, strengthens the precursors required for resveratrol synthesis, and greatly increases the yield of the product resveratrol.

The application discloses a kind of p-coumaric acid or p-coumaric acid derivative production level improved microorganism and its construction method and application, comprising: the genetic target point related to p-coumaric acid metabolic pathway of microorganism is reformed, target point is 4-coumaroyl coenzyme A ligase, hydroxycinnamoyl coenzyme A hydratase lyase, hydroxyacyl coenzyme A dehydrogenase, beta-ketoacyl-coa thiolase, 3- (aryl) propenoate reductase one or more than two combinations, reformation is removed the partial activity or all activity of p-coumaric acid metabolic pathway related genetic target point.The application reduces the degree of degradation of p-coumaric acid by microorganism by reformation p-coumaric acid metabolic pathway related genetic target point, to improve the yield and / or production efficiency of p-coumaric acid and p-coumaric acid derivative produced by microorganism.

The invention belongs to the field of gene engineering, and particularly relates to a glechoma longituba 4-coumaric acid-coenzyme A ligase gene Gl4CL as well as an encoded product and application thereof. The glechoma longituba 4-coumaric acid coenzyme A ligase gene Gl4CL is cloned, prepared and obtained from glechoma longituba medicinal plants for the first time, the nucleotide sequence of the gene is shown as SEQ ID NO.1, and the amino acid sequence of a product coded by the nucleotide sequence is shown as SEQ ID NO.2. The glechoma longituba 4-coumaric acid coenzyme A ligase gene Gl4CL is a key regulatory gene in a glechoma longituba flavonoid biosynthetic pathway, and the ligase can be applied to the process of preparing p-coumaryl coenzyme A by taking p-coumaric acid as a substrate, so that the biosynthesis of a flavonoid compound is further promoted. The invention can provide an effective method for meeting the huge market demand faced by the flavonoid compound.

The invention relates to the field of gene engineering, in particular to saccharomyces cerevisiae gene engineering bacteria for producing verbascoside and echinacoside through de novo synthesis and application of the saccharomyces cerevisiae gene engineering bacteria. The genetically engineered bacterium contains a coded chorismate synthase mutant, a chorismate mutase mutant, shikimic acid kinase, cytochrome b5, glucose-6-phosphate-dehydrogenase, aromatic acetaldehyde synthase, UDP (User Datagram Protocol) glucosyltransferase, tyrosine ammonia lyase, phenylalanine ammonia lyase, cinnamate hydroxylase, 4-coumaric acid: coenzyme A ligase and acyltransferase. The invention relates to a gene encoding a UDP glucosyltransferase mutant, and more preferably, the gene encoding the UDP glucosyltransferase mutant comprises genes of a UDP glucosyltransferase mutant, a UDP glucosyltransferase mutant, a UDP glucosyltransferase mutant and a UDP glucosyltransferase mutant. Under a shake flask condition, verbascoside produced by the strain reaches 231 mg / L, echinacoside produced by the strain reaches 184 mg / L, and the strain has industrial application potential.

The invention relates to the technical field of biological materials, in particular to engineering modified bacteria for producing butyrolactam. The engineering modified bacterium heterologously expresses a butyrolactam synthesis gene, wherein the butyrolactam synthesis gene is selected from the group consisting of a beta-alanine coenzyme A transferase coding gene, a carnitine coenzyme A ligase coding gene and an acyl coenzyme A ligase coding gene. The engineering modified bacterium provided by the embodiment of the invention has the advantages of high yield, low cost and the like when being used for synthesizing metabolites such as butyrolactam, a low-cost production platform for co-production of butyrolactam or butyrolactam and other products can be realized, and intracellular or extracellular products can be efficiently produced under non-sterilization, open and continuous fermentation conditions.

The invention relates to the technical field of genetic engineering and bioengineering, in particular to a surfactin-producing genetic engineering bacterium and application thereof. A genome of the genetically engineered bacterium comprises the following components; an expression cassette encoding a T7 RNA polymerase; an expression cassette of the surfactin synthesis gene cluster srfAABCD; and an expression cassette of the long-chain fatty acid-coenzyme A ligase gene lcfA; the expression cassette of the surfactin synthetic gene cluster srfAABCD and the expression cassette of the long-chain fatty acid-coenzyme A ligase gene lcfA also comprise a T7 promoter. According to the method, the surfactin is efficiently synthesized by the bacillus subtilis, the final shake flask yield reaches 19.3 g / L or above, and the method has huge industrial application potential.

The invention discloses a microorganism capable of improving the production level of p-coumaric acid or p-coumaric acid derivatives as well as a construction method and application of the microorganism. The construction method comprises the following steps: transforming genetic targets related to metabolic pathways of p-coumaric acid of the microorganism; the target spot is one or a combination of two or more of 4-coumaroyl coenzyme A ligase, hydroxycinnamyl coenzyme A hydratase lyase, hydroxyacyl coenzyme A dehydrogenase, beta-keto-acyl coenzyme A thiolated enzyme and 3-(aryl) acrylate reductase, and is transformed to remove part or all of the activity of genetic target spots related to a coumaric acid metabolic pathway. Genetic targets related to metabolic pathways of p-coumaric acid are modified, so that the degree of degrading p-coumaric acid by microorganisms is reduced, and the yield and / or production efficiency of p-coumaric acid and p-coumaric acid derivatives produced by the microorganisms are / is improved.

The application discloses a kind of resveratrol engineering bacteria synthesized by p-coumaric acid, construction and application, belong to biotechnology field.The application is by expressing the coding gene of 4-coumaroyl coenzyme A ligase mutant and / or the coding gene of resveratrol synthase mutant and the coding gene of acetyl-coa carboxylase and / or the coding gene of acetyl-coa synthase, on the basis of significantly improving the catalytic activity of 4-coumaroyl coenzyme A ligase, resveratrol synthase respectively, by the enhanced expression of acetyl-coa, malonyl-coa, the obtained Yarrowia lipolytica engineering bacteria can efficiently utilize p-coumaric acid to produce resveratrol, the yield reaches 2.4g / L in flask fermentation, substrate conversion rate reaches 96.3% in 5L fermenter batch feeding fermentation, and the yield of resveratrol reaches 47.8g / L.

The invention discloses a saccharomyces cerevisiae engineering strain for reducing accumulation of naringenin and phloretin and efficiently producing phloretin and application of the saccharomyces cerevisiae engineering strain, and belongs to the technical field of microbial genetic engineering. According to the invention, the naringenin chalcone reductase gene MdNCR from apples is expressed in saccharomyces cerevisiae for the first time, and a new way for directly synthesizing phloretin from naringenin chalcone is constructed, so that the phloretin yield of the strain is increased by over 300% compared with that of Y22, the naringenin accumulation is reduced by more than 85%, and the problem of carbon metabolic flux shunting is effectively solved; 4-coumaroyl coenzyme A ligase and chalcone synthase are synergistically optimized aiming at a large amount of accumulation of the phloreic acid after introduction of an exogenous PAL pathway, and a'phloreic acid recovery cycle 'is constructed to re-introduce the phloreic acid into a synthesis main pathway; finally, the phloretin content of the engineering strain Y2226 is reduced to 22.3 mg / L, the phloretin yield is increased to 110.7 mg / L, and a solution is provided for accumulation of phloretin by-products synthesized by microorganisms.

The invention provides an engineering bacterium for synthesizing phloretin as well as a construction method and application of the engineering bacterium. The engineering bacteria comprise escherichia coli and genes which are co-expressed in the escherichia coli and are used for coding tyrosine deaminase, p-coumaroyl-coenzyme A ligase, chalcone synthase and reductase, the tyrosine deaminase is derived from rhodotorula or hypericum monogynum, the p-coumaroyl-coenzyme A ligase is derived from arabidopsis A.thaliana, petunia or apple, and the chalcone synthase is derived from chalcone synthase. The chalcone synthase gene is derived from petunia hybrida, and the reductase gene is derived from clostridium or acetyl clostridium. Efficient synthesis of phloretin can be realized by utilizing the engineering bacteria, and the yield of the phloretin synthesized from the beginning can reach 244.55 mg / L. The invention also provides an application of the engineering bacterium in biosynthesis of phloretin derivatives such as 3-hydroxy phloretin, hesperetin dihydrochalcone, hesperetin dihydrochalcone glucoside, neohesperidin dihydrochalcone and the like.