35 results about "Tyrosine ammonia-lyase" patented technology

The present invention provides several methods for biological production of para-hydroxycinnamic acid (PHCA). The invention is also directed to the discovery of new fungi and bacteria that possess the ability to convert cinnamate to PHCA. The invention relates to developing of a new biocatalyst for conversion of glucose to PHCA by incorporation of the wild type PAL from the yeast Rhodotorula glutinis into E. coli underlining the ability of the wildtype PAL to convert tyrosine to PHCA. The invention is also directed to developing a new biocatalyst for conversion of glucose to PHCA by incorporation of the wildtype PAL from the yeast Rhodotorula glutinis plus the plant cytochrome P-450 and the cytochrome P-450 reductase into E. coli. In yet another embodiment, the present invention provides for the developing of a new biocatalyst through mutagenesis of the wild type yeast PAL which possesses enhanced tyrosine ammonia-lyase (TAL) activity.

A recombinant micro-organism producing resveratrol by a pathway in which phenylalanine ammonia lyase (PAL) produces trans-cinnamic acid from phenylalanine, cinnamate 4-hydroxylase (C4H) produces 4-coumaric acid from said trans-cinnamic acid, 4-coumarate-CoA ligase (4CL) produces 4-coumaroyl CoA from said 4-coumaric acid, and resveratrol synthase (VST) produces said resveratrol from said 4-coumaroyl CoA, or in which L-phenylalanine- or tyrosine-ammonia lyase (PAL / TAL) produces 4-coumaric acid, 4-coumarate-CoA ligase (4CL) produces 4-coumaroyl CoA from said 4-coumaric acid, and resveratrol synthase (VST) produces said resveratrol from said 4-coumaroyl CoA. The micro-organism may be a yeast, fungus or bacterium including Saccharomyces cerevisiae, E. coli, Lactococcus lactis, Aspergillus niger, or Aspergillus oryzae.

A recombinant micro-organism producing resveratrol by a pathway in which phenylalanine ammonia lyase (PAL) produces trans-cinnamic acid from phenylalanine, cinnamate 4-hydroxylase (C4H) produces 4-coumaric acid from said trans-cinnamic acid, 4-coumarate-CoA ligase (4CL) produces 4-coumaroyl CoA from said 4-coumaric acid, and resveratrol synthase (VST) produces said resveratrol from said 4-coumaroyl CoA, or in which L-phenylalanine- or tyrosine-ammonia lyase (PAL / TAL) produces 4-coumaric acid, 4-coumarate-CoA ligase (4CL) produces 4-coumaroyl CoA from said 4-coumaric acid, and resveratrol synthase (VST) produces said resveratrol from said 4-coumaroyl CoA. The micro-organism may be a yeast, fungus or bacterium including Saccharomyces cerevisiae, E. coli, Lactococcus lactis, Aspergillus niger, or Aspergillus oryzae.

The invention relates to a biocatalytic method for producing p-vinylphenols, comprising a three-stage one-pot reaction according to the following reaction scheme: (A) wherein a) an optionally substituted phenol (1) is bound to pyruvic acid (BTS) to form the optionally substituted tyrosine (2) by means of catalytic action of a tyrosine phenol-lyase (TPL) and in the presence of ammonium ions, b) ammonia is eliminated from the tyrosine (2) by means of catalytic action of a tyrosine-ammonia-lyase (TAL) or phenyl-ammonia-lyase (PAL), in order to produce an optionally substituted p-coumaric acid (3), and c) the p-coumaric acid (3) undergoes a decarboxylation by means of catalytic action of a phenolic acid decarboxylase (PAD), in order to produce the desired, optionally substituted p-vinyl phenol(4); d) wherein the resultant CO2 is removed from the reaction system, in order to move the chemical equilibrium of all three reaction steps in the direction of the products.

The invention discloses an engineering bacterium, and an application thereof in the production of caffeic acid, and belongs to the technical field of bioengineering. The engineering bacterium providedby the invention is a recombinant bacterium capable of producing caffeic acid at a low cost; the recombinant bacterium can simultaneously express four enzymes which are tyrosine phenol lyase, tyrosine ammonia lyase, L-lactate dehydrogenase and NADH oxidase respectively; and the recombinant bacterium knocks out a phenolic substance-decomposing gene, and can achieve enhanced expression of any one or more of a lactic acid transporter gene, a catechol transporter gene and a coenzyme synthesis-related gene. The engineering bacterium has the advantages of realization of the efficient production ofcaffeic acid, simple process, few impurities and great industrial application values.

The present invention provides engineered tyrosine ammonia-lyase (TAL) polypeptides and compositions thereof. In some embodiments, the engineered TAL polypeptides have been optimized to provide enhanced catalytic activity while reducing sensitivity to proteolysis and increasing tolerance to acidic pH levels. The invention also provides methods for utilization of the compositions comprising the engineered TAL polypeptides for therapeutic and industrial purposes.

The invention relates to a genetically engineered bacterium for producing ferulic acid and a biosynthesis method of the ferulic acid, and belongs to the technical field of biological application. Thegenetically engineered bacterium for producing the ferulic acid can produce the ferulic acid by using tyrosine and methionine as raw materials. According to the biosynthesis way of the ferulic acid inplants, the recombinant genetically engineered bacterium is obtained by a genetic engineering method, and the genetically engineered bacterium can heterologously express tyrosine ammonia lyase, coumaric acid-3-hydroxylase and caffeic acid-3-O-methyltransferase, so that tyrosine biotransformation is promoted, the ferulic acid is produced by one-pot fermentation, the conversion rate is high, the purity of the obtained ferulic acid is high, the process is simple and clean, the cost is low, and the genetically engineered bacterium for producing the ferulic acid and the biosynthesis method of theferulic acid can be used for large-scale industrial production.

The invention discloses a method for producing eriodictyol by reforming escherichia coli in metabolic engineering, belonging to the field of metabolic engineering. The step of transforming naringenin into the eriodictyol is successfully realized through a gene engineering technology by carrying out fusion expression on two genes, namely a flavone 3' hydroxylase gene tF3'H with a membrane binding sequence cut and a P450 reductase gene tCPR, in escherichia coli. A tyrosine ammonia lyase gene TAL from R.glutinis, 4-coumarate:coenzyme A ligase gene 4CL from P.crispum, a chalcone synthase gene CHS from P.hybrida and a chalcone isomerase gene CHI from M.sativa are simultaneously expressed to realize that naringenin is directly produced through tyrosine, so that an engineered strain for directly producing eriodictyol through tyrosine is established. According to the method, in order to increase the output of eriodictyol, an acetokinase ackA gene contained in an escherichia coli genome is removed by being knocked, an acetyl coenzyme A synthetase gene acs and an acetyl coenzyme A carboxylase gene ACC are excessively expressed, and the output of eriodictyol can finally reach 106.7 mg / L.

The invention relates to a strain for producing tyrosine ammonia lyase and an application of the strain, and belongs to the field of biotechnology. The strain is classified and named as Acinetobacterjohnsonii, is preserved in the China General Microbiological Culture Collection Center on May 15, 2020, and has a preservation number of CGMCC No.19831. The strain provided by the invention has good hereditary stability and low culture cost, the generated tyrosine ammonia lyase has high enzyme activity, and the strain has a wide applicable substrate spectrum and can be directly used for direct conversion and synthesis of p-hydroxycinnamic acid and p-hydroxycinnamic acid derivatives. During catalytic synthesis, addition of a surfactant, an organic solvent and other membrane penetrating agents is not needed, introduction of exogenous impurities is avoided, extraction and separation operation is facilitated, and advantages are achieved for industrial application.

The invention discloses a biological preparation method of caffeic acid, which belongs to the technical field of microorganisms. The present invention selects high osmotic pressure Candida glycerol producing yeast CCTCC M93018 to construct engineering bacteria, knocks out L-tryptophan synthesis gene trp1 and L-phenylalanine synthesis gene pheA, and overexpresses 3-deoxy-D-arabinan Keto-7-phosphate synthase gene aro4 and bifunctional choroidate synthase and flavin reductase gene aro7, and tyrosine ammonia lyase, and 4-hydroxyphenylacetic acid-3-monooxygenase Mutation, in the yeast to achieve de novo synthesis of caffeic acid with glucose as carbon source, caffeic acid production reached 87.6mg / L. The preparation method of the invention has the characteristics of convenient operation, high conversion efficiency, low production cost, wide industrial application prospect and the like.

The invention discloses an engineering bacterium and its application for producing caffeic acid, and belongs to the technical field of biological engineering. The invention provides a recombinant bacteria capable of producing caffeic acid at low cost; the recombinant bacteria simultaneously expresses four enzymes, namely tyrosine phenol lyase, tyrosine ammonia lyase, L-lactate dehydrogenase, and NADH oxidase; Further, the recombinant bacteria of the present invention also knocks out the phenolic substance decomposition gene, and strengthens the expression of any one or more of the lactic acid transport gene, the catechol transport gene, and the coenzyme synthesis-related gene. The invention realizes the efficient production of caffeic acid, and has the advantages of simple process and few impurities, and has important industrial application value.

The invention discloses a novel method for synthesizing danshensu. The method uses tyrosine ammonia lyase, phenylpyruvate reductase and glucose dehydrogenase to catalyze levodopa to produce danshensu. Specifically, 0.5-1.5mol of levodopa (3,4-dihydroxyphenylalanine), 10-30g / L of wet bacterium of tyrosine ammonia lyase, and 10-30g of wet bacterium of phenylpyruvate reductase / L, 10-30g / L of wet cells of glucose dehydrogenase were placed in a reactor and mixed evenly, the pH value was adjusted to 7.0, the temperature was 30°C, and the catalytic reaction was carried out for 16-20h to obtain Danshensu. For the first time, the method uses tyrosine ammonia lyase, phenylpyruvate reductase and glucose dehydrogenase to produce danshensu enzymatically with levodopa as a substrate, saving raw materials and avoiding the pollution of chemical synthesis.

The invention relates to a strain producing tyrosine ammonia-lyase and its application, which belongs to the field of biotechnology. The strain is classified and named as Acinetobacter johnsonii, and has been preserved in China's microbial strains on May 15, 2020 General Microbiology Center of the Preservation Management Committee, deposit number: CGMCC No.19831. The bacterial strain provided by the present invention has good genetic stability, low cultivation cost, high enzymatic activity of tyrosine ammonia-lyase, and wide applicable substrate spectrum. There is no need to add membrane-permeable agents such as surfactants and organic solvents during synthesis, avoiding the introduction of foreign impurities, which is conducive to extraction and separation operations, and has advantages in industrial applications.

The invention discloses a biological method for producing resveratrol, which comprises the following steps: ligating target genes obtained by restriction enzyme digestion to an expression vector, wherein the target genes include the sequences of phenylalanine hydroxylase (PAH), tyrosine ammonia lyase (TAL) 4-coumarate:coenzyme A ligase (4CL) and resveratrol synthase (RS); transforming the constructed expression vector into a strain to obtain a recombinant engineered strain; and fermenting the recombined engineered strain. Compared with the prior art, the technical scheme provided by the invention uses the genetically engineered strain for fermentation to produce resveratrol, so as to realize denovo synthesis of resveratrol with no need of adding substrates. The method disclosed by the invention solves the source problem of resveratrol, and at the same time, reduces the production cost in a maximum extent, and is beneficial for industrial production.

The present invention relates to the field of genetic engineering, in particular to a rosavir analog, a genetically engineered bacterium producing the rosavir analog, and an application thereof. The rosevin analog has a structure shown in formula I, formula II, formula III or formula IV, and the genetically engineered bacterium contains phenylalanine ammonia lyase or tyrosine ammonia lyase, 4-coumaric acid coenzyme A Genes for ligase 4CL, cinnamoyl-CoA reductase CCR, and UDP glucosyltransferase UGT. The genetically engineered bacteria can ferment and produce Triandrin and Triandrin B (Formula IV); or Triandrin, Triandrin B, Triandrin C (Formula II) and Triandrin D (Formula III), and the yield of Triandrin and Triandrin B can reach up to 1.7g / L and 1.6g / L; or Luosaiwei B (formula I), the highest output can reach 1.6g / L.