11 results about "Malate dehydrogenase" patented technology

Provided is a genetically engineered strain of Saccharomyces cerevisiae ST100-Suc3 for producing succinic acid. The Saccharomyces cerevisiae ST100-Suc3 is obtained by overexpressing malate dehydrogenase (ScMDH3R), fumarase (RoFUM) and fumarate reductase (TbFRD) in Saccharomyces cerevisiae PY-EDS8 while knocking out a glycerol-3-phosphate dehydrogenase (GPD1) gene, an alcohol dehydrogenase (ADH1) gene and an aldehyde dehydrogenase (ALD6) gene. The Saccharomyces cerevisiae strain is Saccharomyces cerevisiae PY-EDS8, with the deposit number GDMCC No: 63815. The strain exhibits tolerance to high-concentration succinic acid and can grow in a liquid medium supplemented with 100 g / L of succinic acid (pH=2.68), but produces almost no succinic acid.

The present invention relates to an improved process for the preparation of 2, 4-dihydroxybutyrate from a carbon source via homoserine, comprising a two-step route: a first step of catalytically converting the primary amino group of homoserine to a carbonyl group by an enzyme having homoserine transaminase activity as defined by E.C.2. 6. 1.1 or E.C.2. 6. 1.42 or E.C.2. 6. 1.57 to obtain 2-oxo-4-hydroxybutyrate, and a second step of catalytically converting the primary amino group of homoserine to a carbonyl group by an enzyme having homoserine transaminase activity as defined by E.C.2. 6. 1.1 or E.C.2. 6. 1.57 to obtain 2-oxo-4-hydroxybutyrate. And-a second step of catalytically reducing the obtained 2-oxo-4-hydroxybutyrate (OHB) to 2, 4-dihydroxybutyrate by an enzyme having OHB reductase activity, the enzyme having OHB reductase activity being a lactic dehydrogenase as defined by E.C. 1.1. 1.27 or E.C. 1.1. 1.28, or a malate dehydrogenase as defined by E.C. 1.1. 1.37, E.C. 1.1. 1.82, or E.C. 1.1. 1.299, and-a second step of catalytically reducing the obtained 2-oxo-4-hydroxybutyrate (OHB) to 2, 4-dihydroxybutyrate by an enzyme having OHB reductase activity, the enzyme having OHB reductase activity being a lactic dehydrogenase as defined by E.C. 1.1. 1.28, or a malate dehydrogenase as defined by and wherein at least one enzyme having aldolase activity, as defined by E.C.4.1. 2.28, is inactivated to prevent degradation of OHB.

The application discloses a succinic acid-producing strain of Issatchenkia orientalis, and construction and application thereof, and belongs to the technical field of microorganisms. Through metabolic engineering, the application knocks out a pyruvate decarboxylase coding gene on a genome of the Issatchenkia orientalis and integrates expression of a pyruvate carboxylase, a malate dehydrogenase, a fumarase and a fumarate reductase, knocks out a glycerol dehydrogenase coding gene on the genome and integrates expression of a dicarboxylic acid outer transport protein from a schizosaccharomyces pombe, simultaneously knocks out carboxylic acid inner transport proteins JEN2-1 and JENE2-2 on the genome, and overexpresses the fumarate reductase and / or the malate dehydrogenase. The modified yeast strain significantly improves a succinic acid yield in a fermentation process, and the succinic acid yield reaches 125.77 g / L, and a production intensity is 1.75 g / L / h. The engineering strain has good industrial application potential in high-efficiency succinic acid production.

The invention relates to the technical field of enzyme engineering, in particular to an expression and purification method of cyanobacteria malate dehydrogenase, which comprises the following steps: S1, transforming a constructed recombinant plasmid into an escherichia coli competent cell; s2, carrying out inoculation and fermentation culture on the bacterial liquid obtained in S1, and collecting thalli; s3, adding the thalli into a buffer solution for cell disruption to obtain a supernatant; s4, adding the supernate into resin, and performing chromatography and elution to obtain purified cyanobacteria malate dehydrogenase protein. The invention lays a foundation for the structure and function research of cyanobacteria MDH. Researches show that the cyanobacteria MDH family possibly has a new characteristic of double-coenzyme dependence, and MDH from part of species of cyanobacteria has good heat resistance and thermal stability and has good industrial application potential.

The application belongs to the technical field of biological medicine, and particularly relates to a sulfonamide compound and a preparation method and application thereof. It is found for the first time that malate dehydrogenase is closely related to aging, and inhibiting malate dehydrogenase can achieve a significant anti-aging effect. Based on this, the application provides a sulfonamide compound, the compound has a significant inhibitory effect on malate dehydrogenase, can prolong the life of nematodes, achieves a significant anti-aging effect, and has good safety, thereby providing more safe compounds for anti-aging drugs.

The invention discloses a malate dehydrogenase (MDH) mutant MaMDHK240A with increased enzyme activity and an application of the malate dehydrogenase (MDH) mutant MaMDHK240A. The invention relates to a malate dehydrogenase mutant MaMDHK240A, and the amino acid sequence of the malate dehydrogenase mutant MaMDHK240A is as shown in SEQ ID NO. 3. According to the mutant MaMDHK240A, site-specific mutagenesis is carried out on an amino acid sequence as shown in SEQ ID NO.2, and 240th Lys is mutated into Ala, so that the activity of the malate dehydrogenase is increased, and the enzyme activity of the malate dehydrogenase is increased by 1.87 times. The malate dehydrogenase mutant MaMDHK240A obtained by the invention can be used as a mutant material capable of remarkably increasing the catalytic activity of the malate dehydrogenase, and provides reference for further research on the functions of the malate dehydrogenase.

The invention relates to the technical field of enzyme engineering, and discloses malic dehydrogenase, engineering pichia kudriavzevii containing the malic dehydrogenase and application of the malic dehydrogenase and the engineering pichia kudriavzevii. In the invention, a pichia kudriavzevii engineering strain with NADH (Nicotinamide Adenine Dinucleotide Hydrocarbon) selection pressure after double knockout in an ethanol / glycerol synthetic route is used as a chassis bacterium, and in-vivo directed evolution screening is carried out to obtain various malate dehydrogenase mutants. The malate dehydrogenase mutants have high activity and can be used as novel efficient tool enzymes for preparing malic acid.

The invention discloses a method for obtaining a high-yield epsilon-polylysine streptomyces diastatochromogenes engineering strain S.diastatochromogenes MDH-OGDH by overexpressing a key enzyme malic dehydrogenase gene MDH and an alpha-ketoglutaric acid decarboxylase gene OGDH in a TCA (Trichoderma Carboxygenase) pathway, the epsilon-polylysine is produced by fed-batch fermentation in a 5L fermentation tank, and the yield (g / L) of the epsilon-polylysine is 137.18% of that of a chassis strain S.diastatochromogenes 6 #-7, and the yield (g / L) of the epsilon-polylysine is 37.18% of that of the chassis strain S.diastatochromogenes 6 #-7. The production intensity (g / L.h) of the strain is 133.68% of that of a chassis strain S.diastatochromogenes 6 #-7; the saccharic acid conversion rate of the strain is 116.04% of that of a chassis strain S.diastatochromogenes 6 #-7, the production cost is remarkably reduced, and an excellent strain is provided for industrial production of epsilon-polylysine.

The present application relates to the technical field of enzyme engineering, and discloses a malate dehydrogenase, an engineered Pichia kudriavzevii containing the same, and application of the engineered Pichia kudriavzevii. In the present application, the Pichia kudriavzevii engineering strain with NADH selection pressure after double knockout of ethanol / glycerol synthesis pathway is used as a chassis strain, in vivo directed evolution screening is carried out, and various malate dehydrogenase mutants are obtained. The malate dehydrogenase mutants have high activity and can be used as a novel and efficient tool enzyme for preparing malic acid.

The invention discloses construction and application of an issatchenkia orientalis strain for producing succinic acid, and belongs to the technical field of microorganisms. Through a metabolic engineering means, a pyruvate decarboxylase coding gene on an issatchenkia orientalis genome is knocked out, and pyruvate carboxylase, malate dehydrogenase, fumarase and fumarate reductase are integrally expressed; a glycerol dehydrogenase coding gene on a genome is knocked out, dicarboxylic acid outer transport protein from schizosaccharomyces pombe is integrally expressed, carboxylic acid inner transport protein JEN2-1 and JENE2-2 on the genome are knocked out at the same time, and fumarate reductase and / or malic acid dehydrogenation are / is overexpressed. The succinic acid yield of the modified yeast strain in the fermentation process is remarkably increased and reaches 125.77 g / L, and the production intensity is 1.75 g / L / h. The engineering strain provided by the invention has good industrial application potential in efficient succinic acid production.

The embodiment of the application belongs to the technical field of malate dehydrogenase preparation, and relates to a thermus thermophilus malate dehydrogenase recombinant gene and a preparation method and application thereof, comprising the following steps: adopting a gene sequence of thermus thermophilus malate dehydrogenase, optimizing the gene sequence of the thermus thermophilus malate dehydrogenase by using the synonymous codon preference of escherichia coli, obtaining a thermus thermophilus malate dehydrogenase recombinant gene, constructing a thermus thermophilus malate dehydrogenase recombinant plasmid based on the thermus thermophilus malate dehydrogenase recombinant gene, transforming the thermus thermophilus malate dehydrogenase recombinant plasmid into escherichia coli and culturing, obtaining a bacteria liquid for expressing malate dehydrogenase, purifying the bacteria liquid for expressing malate dehydrogenase, and obtaining malate dehydrogenase. The malate dehydrogenase expressed by the application has high yield, short production cycle and high activity.