150results about "Carbon-nitrogen lyases" patented technology

The invention belongs to the technical field of genetic engineering and enzyme engineering, and particularly relates to a genetically engineered bacterium for stably and efficiently producing L-isoleucine as well as a construction method and application of the genetically engineered bacterium. The genetically engineered bacterium takes E. coliTHRD as a host and contains an RNA (Ribonucleic Acid) polymerase beta-subunit mutant coding gene rpoBM; compared with a parent RNA polymerase beta-subunit, the RNA polymerase beta-subunit mutant has the advantage that the amino acid residue at the 618th site of the amino acid sequence is sequentially replaced with leucine from the N terminal to the C terminal. The genetically engineered bacterium disclosed by the invention can tolerate L-isoleucine and accumulate less alpha-ketobutyric acid, and has a very good industrial application prospect.

The invention discloses a yarrowia lipolytica engineering strain with high yield of resveratrol as well as construction and application of the yarrowia lipolytica engineering strain, and belongs to the technical field of synthetic biology and metabolic engineering. The invention provides a yarrowia lipolytica engineering strain. A 4-coumaric acid-CoA ligase gene, a multi-copy resveratrol synthase gene, a transketolase 1 gene, a fusion gene of a histidine phosphate aminotransferase gene and a tyrosine ammonia lyase gene, a glycolytic pathway hexokinase gene, a 6-phosphofructokinase-1 gene and a phosphoglycerate kinase gene are integrated, so that the recombinant protein is obtained. According to the present invention, by using glucose as the substrate, the high yield of resveratrol can be achieved by using glucose as the substrate, the resveratrol yield during shake-flask culture fermentation can achieve 3.157 g / L, the resveratrol yield can be increased to 30.7 g / L during 5L-scale fed-batch fermentation, and the resveratrol yield can be increased to 30.7 g / L when the resveratrol yield is increased to 30.7 g / L when the resveratrol yield is increased to 30.7 g / L when the resveratrol yield is increased to 30.7 g / L; the maximum value is reported by the yarrowia lipolytica system at present.

The invention relates to the technical field of synthetic biology and microbial engineering, in particular to recombinant escherichia coli for de novo synthesis of forsythiaside A. The invention provides recombinant escherichia coli for de novo synthesis of forsythiaside A. A preparation method of the recombinant escherichia coli comprises a step of improving an escherichia coli starting strain, and the escherichia coli starting strain is escherichia coli MG1655. In the fermentation process of the recombinant escherichia coli, effective accumulation of forsythiaside A in fermentation liquor can be achieved, and a foundation can be laid for subsequent construction of high-yield forsythiaside A engineering bacteria.

The invention provides a strain for producing L-isoleucine as well as a construction method and application thereof. The strain is obtained by modifying a chassis strain escherichia coli XX12 by utilizing a metabolic engineering modification method, ldhA, adhE and pflB genes are deleted, the transcriptional levels of aspC, pykF, pntAB, ppK, ppc, ilvAfbr, ilvIHfbr and ygaZH genes are up-regulated, the transcriptional level of a leuA gene is down-regulated, a bcd gene derived from B.subtilis 168 and ppnK and cysK genes derived from Cornebacterium glutamicum ATCC 13032 are heterologously expressed, and the strain has the advantages that the strain can be used for producing L-isoleucine; the method has the advantages of no need of adding resistant substances, good L-isoleucine synthesis capability, short fermentation period by using glucose as a carbon source, and good economic benefit and industrial application value.

The invention discloses a method for producing p-coumaric acid through double-enzyme cascade catalysis, which comprises the following steps: taking phenol, pyruvic acid and ammonium chloride as substrates, and carrying out cascade enzyme catalytic reaction on a tyrosine phenol lyase product and a tyrosine ammonia lyase product to synthesize p-coumaric acid. According to the method, phenol, pyruvic acid and ammonium chloride are used as substrates, a tyrosine phenol lyase product and a tyrosine ammonia lyase product are used as catalysts, the tyrosine phenol lyase product can convert the substrates into L-tyrosine, and the tyrosine ammonia lyase product can directly use the L-tyrosine as the substrate to convert the L-tyrosine into p-coumaric acid. Therefore, the tyrosine phenol lyase product and the tyrosine ammonia lyase product can realize cascade enzyme catalytic reaction in the same reactor, phenol with lower cost can be adopted as a conversion substrate, separation and purification of an intermediate product L-tyrosine are avoided, and the whole process flow is simple and efficient.

The invention relates to the technical field of synthesis of natural dyes by a biological method, in particular to an antibiotic-demand-free ornamental blue high-yield strain as well as construction and application of the antibiotic-demand-free ornamental blue high-yield strain. Corynebacterium glutamicum is used as an initial strain, a tricarboxylic acid circulating gene is regulated and interfered, and the copy number of exogenous blue pigment synthetase BpsA in a genome is adjusted, so that the ornamental blue high-yield strain is obtained. The strain does not need to use antibiotics in the fermentation process, the yield is stable, the product production capacity equivalent to the level when antibiotics are used can still be achieved through fermentation under the antibiotic-free condition, and the growth performance of the strain is kept. The method for producing the ornamental blue by using the strain provided by the invention ensures that the ornamental blue product is natural and green, also improves the efficient production capacity of the strain, solves the contradiction between green environmental protection and stable high yield of the ornamental blue dye, is green and environment-friendly, and has a remarkable industrial application prospect.

The invention provides a tyrosine ammonia lyase mutant and application thereof in production of p-coumaric acid. The tyrosine ammonia lyase mutant is obtained by mutating wild type tyrosine ammonia lyase with an amino acid sequence as shown in SEQ ID No.1, and a mutation site is selected from at least one of Q108S, Y340Q and P375H. The catalytic activity center of wild type tyrosine ammonia lyase is analyzed, and site-directed mutagenesis is carried out on five catalytic activity center sites, so that the mutant shows different enzyme catalytic activity when the mutation sites are different, the mutation sites are the same but the mutation targets are different; q108S, Y340Q and P375H single mutations can significantly improve the enzyme catalytic activity of the tyrosine ammonia lyase mutant, and the tyrosine ammonia lyase mutant having any two mutation sites, especially having three mutation sites, also has better enzyme catalytic activity than the wild type, and has great application potential.

The invention relates to the technical field of bioengineering, and discloses a phenylalanine ammonia lyase mutant and application thereof. In order to improve the synthesis efficiency of the L-4, 4 '-biphenyl alanine derivative, single-point or multi-point combined substitution is carried out on the 96th amino acid, the 97th amino acid and the 410th amino acid of the phenylalanine ammonia lyase sourced from Lactobacillus bacterium to obtain mutants, and the mutants are single mutants F96V and L97V, and double-site combined mutants F96V / I410V and L97V / I410V. Compared with phenylalanine ammonialyase from other sources, the phenylalanine ammonialyase mutant provided by the invention can accept a large-volume (E)-4-phenylcinnamic acid substrate, has higher catalytic conversion rate, wider substrate range and higher yield, is lower in production cost, and is suitable for industrial production. And the large-volume biphenyl alanine compound can be asymmetrically prepared with extremely high stereoselectivity.

The invention provides a composition comprising a Plasmodium species multimeric protein capable of forming a nanoparticle and at least one antigen of interest, wherein the multimeric protein comprises a monomer chosen from Plasmodium species pyridoxal 5'-phosphate synthase (PLP), chaperone 60 protein (Cpn60), and caseinolytic protease (Clp). In embodiments, the invention provides the composition wherein the monomers assemble to form a multimeric nanoparticle and / or which comprises an adjuvant. The invention further provides a nucleic acid encoding the multimeric protein and the at least one antigen of interest, E. coli comprising the nucleic acid, and the use of such in a method for producing the inventive multimeric protein and the at least one antigen of interest. Also provided are a method and use of the inventive multimeric protein and the at least one antigen of interest for immunizing a subject against Plasmodium species.

The invention discloses efficient synthesis of guanidinoacetic acid by a carbamyl phosphoric acid synthesis-enhanced multi-enzyme cascade system, and belongs to the field of biological catalysis engineering. The invention provides a dominant mutant E31K / G351N of L-arginine: glycine amidino transferase, the catalytic efficiency of guanidinoacetic acid is improved by 34.6% compared with that of wild type, an eight-enzyme synergistic arginine circulation system is subsequently constructed based on a mutant strain, 18.35 g / L GAA (56.75 mM) is realized by a 5L fermentation tank, the arginine conversion rate reaches 261.12%, and the yield of the glycine amidino transferase is greatly improved. The highest level of catalytic synthesis of guanidinoacetic acid by escherichia coli is publically reported at home and abroad at present. According to the research, an efficient technical route is provided for GAA industrial production, and a universal strategy framework is established for rational design of a multi-enzyme system and multi-gene co-expression optimization.

This invention provides compositions and treatment methods for reducing hyperphenylalaninemia. [Solution] A genetically modified bacterium is provided, comprising: a) one or more genes encoding phenylalanine ammonia lyase (PAL), which are operably linked to a promoter that is not naturally associated with the PAL gene and can be directly or indirectly induced; b) one or more genes encoding a phenylalanine transporter, which are not naturally associated with the phenylalanine transporter gene and can be operably linked to a promoter that is directly or indirectly induceable; and c) one or more genes encoding mutant fumarate and nitrate reductase (FNR), which are not naturally associated with the FNR gene and can be operably linked to a promoter that is directly or indirectly induceable.

The invention discloses a Rogowski eutrophy alkali-producing engineering strain for producing p-coumaric acid as well as a construction method and application of the Rogowski eutrophy alkali-producing engineering strain. The preparation method comprises the following steps: by taking a chemoautotrophic microorganism Ropriavidus alcaligenes H16 as an original strain, firstly, by knocking out a synthetic route of a strain poly-3-hydroxybutyrate (PHB), introducing a tyrosine ammonia lyase gene from Rhodotorula glutinis to construct a basic engineering strain capable of producing p-coumaric acid, and then, carrying out fermentation on the basic engineering strain to obtain the p-coumaric acid. Then, endogenous 3-deoxy-7-phosphoheptanone acid synthase gene aroG1 and chorismate synthase gene aroC are overexpressed, and the total copy is increased, so that the p-coumaric acid synthetic pathway flux of the engineering strain is enhanced; and introducing NAD (P) transhydrogenase gene pntAB from Escherichia coli to improve the supply of cofactors, so as to finally obtain the eutrophy alcaligenes Rosei engineering strain capable of synthesizing p-coumaric acid by using CO2.

The present application relates to the technical field of synthetic biology, in particular to a multi-module synergistic optimization of brown cyanidin and brevifolin production of saccharomyces cerevisiae engineering bacteria and its construction method and application. The saccharomyces cerevisiae engineering bacteria includes naringenin synthesis pathway, optimization pathway of shikimic acid, synergistic pathway of phenylalanine, 6-OH luteolin synthesis pathway, brown cyanidin and brevifolin synthesis pathway, multiple copies of SbF6H and MpalOMT2, NADPH optimization pathway, and SAM cycle optimization pathway. The saccharomyces cerevisiae engineering bacteria is used for producing brown cyanidin and brevifolin, and improving the yield of the two.

The present invention concerns a microorganism comprising at least one heterologous gene coding for a L-arginine:glycine amidinotransferase (AGAT) and 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) and a method for the fermentative production of guanidinoacetic acid (GAA) using such microorganism. The present invention also relates to a method for the fermentative production of creatine.

The invention relates to the technical field of medicinal chemistry, and particularly discloses application of echinacoside in preparation of a medicine for improving autism social and cognitive impairment, and the application comprises the following steps: constructing a recombinant saccharomyces cerevisiae strain, and knocking out a precursor competition pathway gene; performing metabolic flow reprogramming fermentation on the recombinant strain; centrifuging the fermentation liquor at 7000-9000 r / min and pretreating the fermentation liquor with an 8-12 kDa ultrafiltration membrane; carrying out dual-targeting affinity chromatography purification on the pretreatment liquid; the preparation method comprises the following steps: preparing echinacoside into a liposome containing pH sensitive poly-histidine; and adding a freeze-drying protective agent into the suspension to carry out gradient pre-freezing and vacuum freeze-drying. According to the application, a mode of combining recombinant saccharomyces cerevisiae strain construction and metabolic flow reprogramming fermentation is adopted, targeted enrichment of echinacoside is realized through affinity chromatography coupled with autism brain region specific double ligands, then the echinacoside is prepared into the liposome containing pH sensitive poly-histidine, the double ligands are modified, and the effects of precise targeting and efficient effect are achieved.

The present invention relates to a microorganism transformed to be capable of producing guanidinoacetic acid (GAA) and containing at least one gene encoding a protein having the function of NADH-dependent dehydrogenase, and a method for the fermentative production of GAA using such a microorganism. The present invention also relates to a method for the fermentative production of creatine.

A method for the incorporation of formaldehyde into biomass is described, comprising the following enzymatically catalyzed steps: (1) condensation of pyruvate and formaldehyde to 4-hydroxy-2-oxobutanoic acid (HOB); (2) amination of the resulting 4-hydroxy-2-oxobutanoic acid (HOB) to produce homoserine; (3) conversion of the resulting homoserine to threonine; (4) conversion of the resulting threonine to glycine and acetaldehyde or acetyl-CoA; (5) condensation of the resulting glycine with formaldehyde to produce serine; and (6) conversion of the resulting serine to produce pyruvate, which can then be used as a substrate in step (1).

Provided herein are phenylalanine-degrading enzyme variants for use in degrading phenylalanine and in treating phenylketonuria (PKU). More specifically, provided herein are phenylalanine-degrading enzyme variants that exhibit increased thermal stability and physicochemical resistance as measured by residual phenylalanine-degrading activity following challenge.

A process for preparing L-glufosinate by the enzymatically catalyzed reaction of homoserine and methylphosphinic acid is provided. The process according to the present disclosure does not include a step of producing O-acyl-L-homoserine, and O-acyl-L-homoserine is not used in the process.

Among the various aspects of the present disclosure is the provision of a biological and biochemical production of piperazic acid derived from the newly discovered production pathway for L-piperazic acid. One aspect of the present disclosure includes a transgenic microorganism (e.g., bacteria) engineered to accumulate piperazic acid and derivatives thereof, including a piperazic acid (Piz)-containing product. Another aspect of the present disclosure includes biochemical and biological methods for producing piperazic acid and derivatives thereof, including a piperazic acid (Piz)-containing product. Another aspect of the present disclosure includes compositions and methods of using isotopically labeled piperazic acid and derivatives thereof, including a piperazic acid (Piz)-containing product.

The invention discloses a separated nucleic acid molecule, a recombinant bacterium and application of the separated nucleic acid molecule to cultivation of a high-sugar tobacco variety, and relates to the technical field of tobacco quality genetic improvement. According to the invention, the NtTD1 gene specifically expressed in the head of the glandular hair is cloned from tobacco, after the NtTD1 gene of the flue-cured tobacco variety K326 is knocked out through a CRISPR / Cas9 gene editing technology, the total sugar content and the reducing sugar content of the flue-cured tobacco leaf of a homozygous mutant tobacco material of the NtTD1 gene are obviously increased, and the development of a tobacco plant is not influenced after the gene is knocked out, so that the content of the total sugar and the reducing sugar in the tobacco leaf is obviously increased. The method has important utilization value in cultivation of high-sugar tobacco varieties, and the method is beneficial to improvement of tobacco leaf quality.

Disclosed in the present invention is the use of a threonine dehydratase in the regulation of the growth of Didymella segeticola and as a bactericide target. The threonine dehydratase gene Td of the present invention is derived from Didymella segeticola, and a knockout mutant is obtained by means of constructing a knockout gene fragment targeting the Td gene and then introducing same into a D. segeticola protoplast. Experiments find that the Td gene plays a role in the growth, the in-vivo pyruvic acid content and the in-vivo ATP content of D. segeticola, and the sensitivity thereof to wuyiencin. By means of molecular docking analysis, molecular dynamics simulation and microscale thermophoresis experiments, it is verified that threonine dehydratase can be used as a bactericide target, and the bactericide wuyiencin is obtained by means of screening. The threonine dehydratase of the present invention can be used as a bactericide target and a key protein in the pathogenic mechanism of a disease, and has broad application prospects in the control of plant pathogenic fungal diseases.

The invention relates to a composition. The present invention relates to a solid carrier, an engineered phenylalanine ammonia lyase or a fragment thereof immobilized on the surface of the solid carrier, and a protective layer for protecting the engineered phenylalanine ammonia lyase or the fragment thereof by embedding the engineered phenylalanine ammonia lyase or the fragment thereof, and a functional component fixed on the surface of the protective layer, in which the functional component fixed on the surface of the protective layer is a polymer comprising repeating units, in which each repeating unit comprises at least one amino group and / or at least one mercapto group. The invention also relates to a method for producing said composition and to the use thereof.

The application provides a tyrosine deaminase mutant and application thereof in production of p-coumaric acid. The tyrosine deaminase mutant is obtained by mutating a wild-type tyrosine deaminase with an amino acid sequence shown in SEQ ID No. 1, and the mutation site is selected from at least one of Q108S, Y340Q and P375H. The application analyzes a catalytic active center of the wild-type tyrosine deaminase, and finds that different mutation sites, same mutation sites but different mutation targets can make the mutant show completely different enzyme catalytic activities, and the single mutation of Q108S, Y340Q and P375H can significantly improve the enzyme catalytic activity of the tyrosine deaminase mutant, and the tyrosine deaminase mutant with any two mutation sites, especially with three mutation sites also has excellent enzyme catalytic activity than the wild type, and has great application potential.

The invention provides apple phenylalanine ammonialyase and application thereof in improving the resistance of apples to bactrocera dorsalis, the amino acid sequence of the apple phenylalanine ammonialyase is SEQ ID NO: 1, and the nucleotide sequence of a coding gene of the apple phenylalanine ammonialyase is SEQ ID NO: 2. The invention further provides a method for improving the bactrocera dorsalis resistance of the apple strain, and according to the method, the content of the apple phenylalanine ammonialyase in the apple strain is increased. According to the apple phenylalanine ammonialyase PAL gene obtained through screening, the larva death rate of an overexpression test group is remarkably higher than that of a blank control group, and it is indicated that overexpression of the PAL gene can possibly affect survival of larvae.

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%.

Provided is a process for preparing L-methionine by enzymatic catalysis reaction of homoserine and methanthiol. The process according to the present disclosure does not comprise a step of generating O-acyl-L-homoserine, and / or O-acyl-L-homoserine is not sued in the process.

Multi-carbon compounds such as ethanol, n-butanol, sec-butanol, isobutanol, tert-butanol, fatty (or aliphatic long chain) alcohols, fatty acid methyl esters, 2,3-butanediol and the like, are important industrial commodity chemicals with a variety of applications. The present invention provides metabolically engineered host microorganisms which metabolize methane (CH4) as their sole carbon source to produce multi-carbon compounds for use in fuels (e.g., bio-fuel, bio-diesel) and bio-based chemicals. Furthermore, use of the metabolically engineered host microorganisms of the invention (which utilize methane as the sole carbon source) mitigate current industry practices and methods of producing multi-carbon compounds from petroleum or petroleum-derived feedstocks, and ameliorate much of the ongoing depletion of arable food source “farmland” currently being diverted to grow bio-fuel feedstocks, and as such, improve the environmental footprint of future bio-fuel, bio-diesel and bio-based chemical compositions.

Provided is a method for preparing glycine by using threonine. In a fermentation process, threonine is decomposed into glycine and acetaldehyde by aldolase. Also provided is a method for simultaneously preparing glycine and acetyl coenzyme A. In a fermentation process, acetaldehyde is reduced into acetyl coenzyme A or an acetyl coenzyme A derivative by acetylating acetaldehyde dehydrogenase; or threonine is dehydrogenated by threonine dehydrogenase to obtain 2-amino-3-ketobutyric acid, which is then ligated by 2-amino-3-ketobutyrate CoA ligase to obtain acetyl coenzyme A, and acetyl coenzyme A can be converted into an acetyl coenzyme A derivative under different fermentation conditions. The present disclosure features a unique concept, enabling the preparation of glycine, acetyl-CoA, and acetyl-CoA derivatives with high conversion efficiency, shorter time consumption, high yield, abundant products, and suitability for industrial processing and production.

The invention provides a biosynthesis method for efficiently producing aromatic non-natural amino acid and site-specific modified recombinant protein, an engineering strain and application of the engineering strain. Specifically, the invention provides an engineering bacterium, and genes of one or more exogenous enzymes selected from the following groups are introduced into the engineering bacterium: threonine aldolase, threonine deaminase and transaminase; and threonine aldolase, threonine deaminase and transaminase are simultaneously expressed in the engineering bacterium. The engineering bacterium disclosed by the invention can be used for efficiently synthesizing aromatic non-natural amino acid through whole-cell catalysis or fermentation conversion. More importantly, the synthetic route provided by the invention can be synergistically coupled with a gene codon extension technology in the same engineering bacterium, and is used for high-efficiency expression of recombinant target protein modified by aromatic non-natural amino acid at a fixed point.