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49 results about "Pyruvate formate lyase" patented technology

In enzymology, formate C-acetyltransferase (pyruvate formate lyase) (EC 2.3.1.54) is an enzyme. Pyruvate formate lyase is found in Escherichia coli and other organisms. It helps regulate anaerobic glucose metabolism. Using radical non-redox chemistry, it catalyzes the reversible conversion of pyruvate and coenzyme-A into formate and acetyl-CoA.

Mutant E. coli strain with increased succinic acid production

A method for isolating succinic acid producing bacteria is provided comprising increasing the biomass of an organism which lacks the ability to catabolize pyruvate, and then subjecting the biomass to glucose-rich medium in an anaerobic environment to enable pyruvate-catabolizing mutants to grow.The invention also provides for a mutant that produces high amounts of succinic acid, which has been derived from a parent which lacked the genes for pyruvate formate lyase and lactate dehydrogenase, and which belongs to the E.coli Group of Bacteria.
Owner:UNIVERSITY OF CHICAGO

XZ-A26 bacterial strain for producing L-alanine with high yield as well as construction method and application of XZ-A26 bacterial strain

The invention discloses an XZ-A26 bacterial strain for producing L-alanine with high yield, which has a preservation number of CGMCC (China General Microbiological Culture Collection Center) No.4036 and has the capacity of generating high-concentration L-alanine through fermentation. The XZ-A26 bacterial strain is constructed by the steps of: integrating an L-alanine dehydrogenase gene on thermophilic fatty bacillus chromosome on lactic dehydrogenase position on an escherichia coli ATCC8739 chromosome, then sequentially knocking out a pyruvate formate lyase gene, an alcohol dehydrogenase gene, an acetokinase gene, a fumaric acid reductase gene and an alanine racemase gene of the escherichia coli chromosome, and then carrying out continuous cell culture in a fermenting tank for obtaining agenetic engineering strain. The invention also relates to a construction method of the XZ-A26 bacterial strain and an application of the XZ-A26 bacterial strain in preparation of the L-alanine. According to the invention, the escherichia coli with the preservation number of CGMCC No.4036 for generating the high-concentration L-alanine through fermentation can be constructed by using a metabolic engineering method, and the yield of the L-alanine generated by using the XZ-A26 bacterial strain reaches up to 115g / L. The XZ-A26 bacterial strain is suitable for industrially producing the L-alanine.
Owner:ANHUI HUAHENG BIOTECH

Genetically engineered bacterium for high-yielding L-valine and method for producing L-valine by fermentation

The invention provides a genetically engineered bacterium for high-yielding L-valine. A construction method of the genetically engineered bacterium comprises the steps that starting from an escherichia coli W3110, an acetolactate synthase gene alsS of a bacillus subtilis is integrated on a genome of the escherichia coli W3110 and subjected to high expression; an escherichia coli ppGpp 3'-pyrophosphoric acid hydrolytic enzyme mutant R290E / K292D gene spoT is integrated on the genome of the escherichia coli W3110 and subjected to high expression; genes of frdA, frdB, frdC and frdD of four subunits of a lactic dehydrogenase gene ldhA, a pyruvate formate lyase I gene pflB and fumaric reductase on the genome of the escherichia coli W3110 are knocked out; a branched chain amino acid transaminasegene ilvE of the escherichia coli is replaced with leucine dehydrogenase gene bcd of the bacillus subtilis; and an acetyl-hydroxyl acid isomerized reductase gene ilvC of the escherichia coli is replaced with an encoding gene of a mutant L67E / R68F / K75E. According to the genetically engineered bacterium for the high-yielding L-valine, an L-valine fermentation method is further modified. Double-phasedissolved oxygen control is adopted, and the L-valine yield and the saccharic acid conversion rate are improved.
Owner:TIANJIN UNIV OF SCI & TECH

Novel rumen bacteria variants and process for preparing succinic acid employing the same

The present invention relates to novel rumen bacterial mutants resulted from the disruption of a lactate dehydrogenase gene (ldhA) and a pyruvate formate-lyase gene (pfl) (which are involved in the production of lactic acid, formic acid and acetic acid) from rumen bacteria; a novel bacterial mutant (Mannheimia sp. LPK7) having disruptions of a lactate dehydrogenase gene (ldhA), a pyruvate formate-lyase gene (pfl), a phosphotransacetylase gene (pta), and a acetate kinase gene (ackA); a novel bacterial mutant (Mannheimia sp. LPK4) having disruptions of a lactate dehydrogenase gene (ldhA), a pyruvate formate-lyase gene (pfl) and a phosphoenolpyruvate carboxylase gene (ppc) involved in the immobilization of CO2 in a metabolic pathway of producing succinic acid; and a method for producing succinic acid, which is characterized by the culture of the above mutants in anaerobic conditions. The inventive bacterial mutants have the property of producing succinic acid at high concentration while producing little or no organic acids, as compared to the prior wild-type strains of producing various organic acids. Thus, the inventive bacterial mutants are useful as strains for the industrial production of succinic acid.
Owner:KOREA ADVANCED INST OF SCI & TECH

Engineering bacteria producing DL-alanine and method of producing DL-alanine by using engineering bacteria

The invention discloses a strain of engineering bacteria producing DL-alanine. Lactic dehydrogenase, pyruvate formate lyase, alcohol dehydrogenase, acetic acid kinase, fumaric acid reductase, alanine racemase and methyl glyoxal synthetase of the strain of engineering bacteria producing the DL-alanine are inactivated; and exogenous L-alanine dehydrogenase gene and alanine racemase gene are integrated on the chromosome of the engineering bacteria. According to the invention, pyroracemic acid, an intermediate product of the glycolysis is converted to L-alanine by integrating the exogenous L-alanine dehydrogenase gene into the chromosome of the engineering bacteria; and an exogenous alanine racemase gene is further integrated into the chromosome, and part of the L-alanine is converted into D-alanine. Then producing the DL-alanine from raw material sugar in one step is realized, the production period of the DL-alanine is decreased and the productivity of the DL-alanine is enhanced.
Owner:ANHUI HUAHENG BIOTECH

Method of improving the efficacy of lactic acid bacterial starter cultures and improved starter culture compositions

Methods of enhancing the growth rate and / or controlling the metabolic activity of lactic acid bacteria and of improving the shelf life and / or the quality of an edible product using lactic acid bacterial organisms which are defective in their pyruvate metabolism. There is also provided starter culture compositions comprising such defective lactic acid bacteria as helper organisms and lactic acid bacterial starter culture strains. Useful helper organisms are Lactococcus strains which are defective with respect to pyruvate formate lyase (Pfl) and / or lactate dehydrogenase (Ldh) activity. The helper organisms may overexpress a gene coding for an NAD+ regenerating enzyme such as NADH oxidase encoded by nox gene.
Owner:CHR HANSEN AS

Ethanol fermentation engineering bacteria capable of reducing fermentation byproduct

The invention relates to an ethanol fermentation engineering bacteria capable of reducing a fermentation byproduct. The ethanol fermentation engineering bacteria is characterized in that a pyruvate formate-lyase gene and a lactate dehydrogenase gene of Escherichia coli are inactivated; and a Zymomonas mobilis gene is transferred into the Escherichia coli. When ethanol is produced by fermenting hexose and pentose with the engineering bacteria, lactic acid and formic acid are not produced and the byproduct is reduced, so that the ethanol productivity is improved.
Owner:THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI

Biocatalyst for production of d-lactic acid (as amended)

A method for producing D-lactic acid in high yield, and to provide a method for producing D-lactic acid with high selectivity, in which optical purity is high and a by-product organic acid is small. In one aspect, a microorganism, wherein activity of pyruvate formate-lyase (pfl) is inactivated or decreased, and further activity of Escherichia coli-derived NADH-dependent D-lactate dehydrogenase (ldhA) is enhanced, is cultured to efficiently produce D-lactic acid. With regard to a method for enhancing ldhA activity, by linking, on a genome, a gene encoding ldhA with a promoter of a gene which controls expression of a protein involved in a glycolytic pathway, a nucleic acid biosynthesis pathway or an amino acid biosynthesis pathway, suitable results are obtained compared to the method for enhancing expression of the gene using an expression vector. A microorganism in which a dld gene is substantially inactivated or decreased is cultured to produce high quality D-lactic acid with reduced concentration of pyruvic acid.
Owner:MITSUI CHEM INC

Novel rumen bacteria variants and process for preparing succinic acid employing the same

The present invention relates to novel rumen bacterial mutants resulted from the disruption of a lactate dehydrogenase gene (ldhA) and a pyruvate formate-lyase gene (pfl) (which are involved in the production of lactic acid, formic acid and acetic acid) from rumen bacteria; a novel bacterial mutant (Mannheimia sp. LPK7) having disruptions of a lactate dehydrogenase gene (ldhA), a pyruvate formate-lyase gene (pfl), a phosphotransacetylase gene (pta), and a acetate kinase gene (ackA); a novel bacterial mutant (Mannheimia sp. LPK4) having disruptions of a lactate dehydrogenase gene (ldhA), a pyruvate formate-lyase gene (pfl) and a phosphoenolpyruvate carboxylase gene (ppc) involved in the immobilization of CO2 in a metabolic pathway of producing succinic acid; and a method for producing succinic acid, which is characterized by the culture of the above mutants in anaerobic conditions. The inventive bacterial mutants have the property of producing succinic acid at high concentration while producing little or no organic acids, as compared to the prior wild-type strains of producing various organic acids. Thus, the inventive bacterial mutants are useful as strains for the industrial production of succinic acid.
Owner:KOREA ADVANCED INST OF SCI & TECH

Novel engineered microorganism producing homo-succinic acid and method for preparing succinic acid using the same

The present invention relates to a mutant microorganism, which is selected from the group consisting of genus Mannheimia, genus Actinobacillus and genus Anaerobiospirillum, producing homo-succinic acid and a method for producing homo-succinic acid using the same, and more particularly to a mutant microorganism producing succinic acid at a high concentration while producing little or no other organic acids in anaerobic conditions, which is obtained by disrupting a gene encoding lactate dehydrogenase (ldhA), a gene encoding phosphotransacetylase (pta), and a gene encoding acetate kinase (ackA), without disrupting a gene encoding pyruvate formate lyase (pfl), as well as a method for producing succinic acid using the same. The inventive mutant microorganism has the property of having a high growth rate and succinic acid productivity while producing little or no organic acids, as compared to the prior strains producing succinic acid. Thus, the inventive mutant microorganism is useful to produce succinic acid for industrial use.
Owner:KOREA ADVANCED INST OF SCI & TECH

Enhancement of Microbial Ethanol Production

A thermophilic microorganism lacks lactate dehydrogenase activity and preferably contains an active pyruvate formate lyase pathway. The thermophilic microorganism contains a gene encoding an NAD-linked formate dehydrogenase. The gene encoding an NAD-linked formate dehydrogenase is preferably a codon optimised version of the gene encoding a thermostable NAD-linked formate dehydrogenase. DNA constructs allow stable expression of the gene encoding an NAD-linked formate dehydrogenase in the thermophilic microorganism. The DNA constructs are based upon use of an insertion sequence to achieve stable expression or recombination to insert the gene encoding an NAD-linked formate dehydrogenase into the lactate dehydrogenase gene, thus achieving gene knockout and new functionality in a single step. The microorganisms are useful in fermentation of sugars to produce ethanol.
Owner:BIOCONVERSION TECH LTD

Genetic engineering strain for producing succinic acid by utilizing glucose and acidogenic fermentation method thereof

InactiveCN102533626AOvercomes the inability to utilize glucoseBacteriaRecombinant DNA-technologyPhosphoenolpyruvate carboxylaseEscherichia coli
The invention belongs to the field of biology engineering technology, and relates to a genetic engineering strain for producing succinic acid by utilizing glucose and an acidogenic fermentation method of the genetic engineering strain. The genetic engineering strain for producing succinic acid by utilizing glucose is named as Escherichia coli BA205 and the preservation number is registered as CCTCC No.M2011447. In the construction process, Escherichia coli which is short of lactic dehydrogenase (LDH) gene and Pyruvate formate-lyase (PFL) gene activity is mainly used as an original strain; phosphoenolpyruvate carboxylase (PPC) gene is removed by utilizing a homologous recombination technology; and phosphoenolpyruvate carboxylase and nicotinic acid phosphoribosyl transferase are excessively co-expressed; therefore the synthesis efficiency of succinic acid is greatly increased. In the fermentation method, a two-stage fermentation manner is adopted, the biomass is improved in an aerobic stage and the acidogenic fermentation is carried out in an anaerobic stage.
Owner:NANJING UNIV OF TECH

Genetic engineering bacterium for synthesizing pyruvic acid and D-alanine as well as construction method and application thereof

The invention discloses a genetic engineering bacterium for synthesizing pyruvic acid and D-alanine as well as a construction method and application thereof. Recombinant escherichia coli of the invention is obtained by knocking out apyruvate dehydrogenase complex achEF gene, a pyruvate lyase pflB gene, a pyruvate oxidase poxB gene, a phosphoenolpyruvate synthase pps gene and a lactic dehydrogenaseldhA gene in escherichia coli BL21 (DE3), blocking a key pathway of further metabolism of pyruvic acid and knocking out genes btsT and cstA for encoding a pyruvic acid transport protein. For the genetic engineering bacterium disclosed by the invention, L-amino acid deaminase pm1 is utilized to catalyze D, L-alanine, so that L-alanine can be converted to the pyruvic acid and can be separated to obtain D-alanine at the same time, and a new idea and method is provided to chiral separation of the D, L-alanine; and the recombinant escherichia coli capable of efficiently synthesizing the pyruvic acid and the D-alanine is obtained through modification, the yield of the pyruvic acid reaches 32.0g / L, the conversion rate of the L-alanine reaches up to 80 percent, and the separation rate of the D / L-alanine reaches up to 80 percent.
Owner:JIANGNAN UNIV

Enterobacteria recombinant strain and use thereof

The invention discloses a recombination strain for an enterobacterium and application of the recombination strain for the enterobacterium. The recombination strain provided by the invention is a recombination strain obtained after deactivation of a pyruvate formate-lyase gene in the enterobacterium. As shown by experiments, the amount of by-product CO2 produced during the fermentation process of the recombination strain is obviously reduced and the amount of target products is obviously improved compared with the control. For example, the amount of lactic acid, the amount of succinic acid and the amount of 2, 3-butanediol produced by fermentation of klebsiella oxytoca are respectively improved by 30 percent, 9 percent and 23 percent compared with the control, and the amount of the CO2 produced is only 8.04 percent compared with the control. The recombination strain helps to release the pressure of the greenhouse effect, has important function in the actual industrial production,not only can greatly reduce the yield of the CO2, but also can improve the conversion rate of converting substrates into products, and has wide application prospect.
Owner:TSINGHUA UNIV

Biocatalyst for production of d-lactic acid

The subject of the present invention is to provide a method for producing D-lactic acid in high yield, and to provide a method for producing D-lactic acid with high selectivity, in which optical purity is high and a by-product organic acid is small. A microorganism, wherein activity of pyruvate formate-lyase (pfl-) is inactivated or decreased, and further activity of Escherichia coli-derived NADH-dependent D-lactate dehydrogenase (ldhA) is enhanced, is cultured to produce a remarkable amount of D-lactic acid in a short time. With regard to a method for enhancing ldhA activity, by linking, on agenome, a gene encoding 1dhA with a promoter of a gene which controls expression of a protein involved in a glycolytic pathway, a nucleic acid biosynthesis pathway or an amino acid biosynthesis pathway, suitable results are obtained compared to the method for enhancing expression of the gene using an expression vector. In addition, a microorganism in which a did gene is substantially inactivatedor decreased is cultured to produce high quality D-lactic acid with reduced concentration of pyruvic acid. Furthermore, it is possible to suppress by-production of succinic acid and fumaric acid whilemaintaining high D-lactic acid productivity by using the above-mentioned microorganism having a TCA cycle, wherein activity of malate dehydrogenase (mdh) is inactivated or decreased, and further activity of aspartate ammonia-lyase (aspA) is inactivated or decreased.
Owner:MITSUI CHEM INC

Enterobacteria recombinant strain and use thereof

The invention discloses a recombination strain for an enterobacterium and application of the recombination strain for the enterobacterium. The recombination strain provided by the invention is a recombination strain obtained after deactivation of a pyruvate formate-lyase gene in the enterobacterium. As shown by experiments, the amount of by-product CO2 produced during the fermentation process of the recombination strain is obviously reduced and the amount of target products is obviously improved compared with the prior art. For example, the amount of lactic acid, the amount of succinic acid and the amount of 2, 3-butanediol produced by fermentation of klebsiella oxytoca are respectively improved by 30 percent, 9 percent and 23 percent compared with the prior art, and the amount of the CO2 produced is only 8.04 percent compared with the prior art. The recombination strain helps to release the pressure of the greenhouse effect, has important function in the actual industrial production, not only can greatly reduce the yield of the CO2 but also can improve the conversion rate of converting substrates into products, and has wide application prospect.
Owner:TSINGHUA UNIV

Engineering bacteria for knocking out pyruvate formate-lyase genes and application of engineering bacteria

The invention discloses engineering bacteria for knocking out pyruvate formate-lyase genes and an application of the engineering bacteria. The pyruvate formate-lyase (flpB) genes in a wild-type strain for producing 1,3-propanediol are knocked out by utilizing a gene homologous recombination and gene insertional inactivation method, so that the gene engineering bacteria with blocked metabolic pathways of methanoic acid can be obtained. The engineering bacteria are used for fermenting production of 1,3-propanediol, and the synthesis of the byproduct methanoic acid is greatly reduced, so that the toxicity effect of the methanoic acid for cells can be reduced, and the concentration, production intensity and substrate conversion rate of the 1,3-propanediol can be improved. The experiment shows that when the engineering bacteria are fermented for 32h in a conventional method, the synthesis amount of the methanoic acid is reduced by more than 90 percent, and the concentration of the 1, 3-propanediol can reach more than 72g / L. By adopting the engineering bacteria, the progress of the technology for producing the 1,3-propanediol in the microorganism fermentation method can be promoted, and the application value can be realized.
Owner:SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI

Increased ethanol production by bacterial cells

Fermentation processes for production of ethanol include supplying a thermophilic microorganism lacking lactate dehydrogenase activity with sugars under conditions in which they metabolise them predominantly by the pyruvate-formate lyase pathway. Importantly, the processes also include supplying sufficient glycerol to convert all of the sugars to ethanol. A further embodiment of the invention includes supplying additional glycerol sufficient to convert the exogenous acetate present in biomass hydrolysates into ethanol. Any type of fermentation system can be used for these processes, but a preferred embodiment includes continuous cultures at high temperatures in which ethanol is removed continuously by vacuum evaporation.
Owner:ENSUS

Metabolically engineered lactic acid bacteria and their use

Mutants of lactic acid bacteria including Lactococcus lactis which are defective in pyruvate formate-lyase production and / or in their lactate dehydrogenase (Ldh) production and methods of isolating such mutants or variants are provided. The mutants are useful in the production of food products or in the manufacturing of compounds such as diacetyl, acetoin and acetaldehyde and as components of food starter cultures. In particular, Lactococcus lactis DN223 deposited under the accession No. DSM 11036.
Owner:CHR HANSEN AS

Use of enzymes which catalyze pyruvate synthesis from formate and acetyl-coa and bacteria expressing same

An isolated microorganism is disclosed being genetically modified to express pyruvate formate lyase (PFL) or 2-ketobutyrate formate lyase, wherein acetyl-CoA of the microorganism is converted to pyruvate in the presence of formate in a single step reaction, wherein the net flux of the reaction is in the direction of pyruvate synthesis.Uses of the microorganism and products comprising same are also disclosed.
Owner:YEDA RES & DEV CO LTD

Escherichia coli for producing isobutanol and ethanol and preparation method of escherichia coli

The invention discloses escherichia coli for producing isobutanol and ethanol and a preparation method of the escherichia coli. The invention provides a method for constructing recombinant bacteria for producing isobutanol and / or ethanol, wherein the method comprises the following steps: 1) expressing an ethanol dehydrogenase adhE and a pyruvate formate-lyase coding gene pflB in the escherichia coli AS108, so as to obtain a recombinant bacterium which is named as A; and 2) reducing enzymatic activity of ethanol dehydrogenase in the recombinant bacterium which is named as A and / or improving enzymatic activity of glucose-6-phosphate dehydrogenase in the recombinant bacterium which is named as A, so as to obtain a recombinant bacterium which is named as B. Experiments prove that excessive NADH can be consumed through a mode of introducing ethanol production in the AS108 strain based upon researches, so that toxicity to cells due to accumulation of the NADH is relieved; and meanwhile, NADPH supply can be promoted by activating a pentose phosphate pathway (PPP), so that an isobutanol production capacity is improved.
Owner:TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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