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30 results about "L-Lactate dehydrogenase" patented technology
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L-lactate dehydrogenase (L-LDH) catalyzes the interconversion of pyruvate and NADH+ to L-lactate and NAD+. H-lactate dehydrogenase (H-LDH) catalyzes the interconversion of D-lactate and ferricytochrome c to pyruvate and ferrocytochrome c. Lactate Dehydrogenase (LDH) is an important enzyme in humans.
The invention relates to an unmarked gene knock-out method of pediococcus acidilactici DQ2 based on homologous recombination. The method comprises the following steps: temperature sensitive-type shuttle plasmid pSET4E and knock-out plasmid containing homologous fragments at upstream and downstream parts of target genes to be knocked out are constructed, the knock-out plasmid is subjected to electrotransformation into pediococcus acidilactici, and single commutators generating homologous recombination for the first time and double-exchange mutant strains generating homologous recombination for the second time are screened and identified. The method disclosed by the invention realizes the unmarked gene knock-out of pediococcus acidilactici for the first time, the obtained knock-out bacterial strain does not carry any resistant gene, can be taken as a original strain for subsequent and reconstruction, and also can be used for large-scale industrial production in a safe mode. The method is used for respective knock-out of L-lactate dehydrogenase gene and d-lactate dehydrogenase gene of the pediococcus acidilactici DQ2 (a preservation number is CGMCC NO.7471), the obtained knock-out bacterial strains are respectively named as pediococcus acidilactici ZP26 and TY112, the preservation numbers are CGMCC NO.8665 and CGMCC NO.8664 respectively, and optically pure D-lactic acid and L-lactic acid are respectively generated.
Yeast includes an introduced gene coding a Homo sapiens- or frog-derived L-lactate dehydrogenase.It is possible to produce lactic acid, which has a variety of applications, efficiently and more cost-effectively by using the yeast and the method of producing lactic acid by using the yeast.
A corynebacterium acetoacidophilum strain and a method for producing succinic acid therefrom belong to the technical field of bioengineering. The invention discloses a corynebacterium acetoacidophilum strain YF / delta ldh and a method for producing succinic acid therefrom. The corynebacterium acetoacidophilum strain is an ldh (L-lactate dehydrogenase) missing bacterium built by gene knockout technology, is preserved in china center for type culture collection on February 29th, 2012, and encoded as CCTCC NO.M2012041. The strain is high in acid yield, highly resistant to sodions and high in foodsafety, can accumulate succinic acid more than 95g / L by being culture for 48 hours in the anaerobic environment using glucose as substrate and sodiumbicarbonate as carbon dioxide donor, and can accumulate succinic acid 136g / L by being cultured for 94 hours.
The invention relates to an isobutanol synthetic bacterium genome dimension metabolic network model and a molecular modification method. According to the method, the necessary path for the thallus growth and the isobutanol synthesis is calculated by a network module, the metabolic network model is subjected to the element mode analysis, the standard deviation coefficient of each gene is calculated, and the isobutanolbiosynthesis yield of different genes is determined; the two key genes including an L-lactate dehydrogenasegene 1dh and a pyruvate dehydrogenase complex E2 subunit coding gene pdhC which are most important to the isobutanol biosynthesis are predicted according to a principle that the standard deviation coefficient is smaller than 0.35, the standard deviation coefficients of the two genes are respectively 2.5 to 3 and 1.5 to 2; and through the determination of the two genes, the isobutanol yield can be improved, and 0.5-0.6C-mol / C-mol glucose can be reached. The key genes which are most important to the isobutanol biosynthesis are obtained through utilizing the relative flux value and are used as modification target spots, the molecular modification of the isobutanol synthetic bacterium is guided, and the isobutanol yield is improved.
The invention provides a sialic acidassay kit, comprising a reagent R1 and a reagent R2, wherein the reagent R1 is prepared from the following components with corresponding concentrations: 8-20g / L tris(hydroxymethyl) aminomethane, 5-15ml / L HCL, 0.5-5KU / L neuraminidase, 0.5-5KU / L lactate dehydrogenase and 0.1-2mmol / L NADH; the reagent R2 is prepared from the following components with correspondingconcentrations: 8-20g / L tris(hydroxymethyl) aminomethane, 5-15ml / L HCL, 0.5-5KU / L N-acetylneuraminic acid aldolase, 25-100g / L sucrose and 0.6-1.2 ml / L triethanolamine. The invention belongs to the technical field of biological detection. The sialic acidassay kit provided by the invention significantly enhances the anti-interference ability while improving the stability, can significantly reducethe interference of bilirubin, hemoglobin, triglyceride and the like, and is good in accuracy when being used for the determination of sialic acid.
The invention discloses an engineering bacterium, and an application thereof in the production of Danshensu by using a cheap substrate, and belongs to the technical field of bioengineering. The engineering bacterium provided by the invention is a recombinant bacterium capable of producing Danshensu at a low cost; the recombinant bacterium can simultaneously express four enzymes which are tyrosinephenol lyase, L-amino acid oxidase, L-lactate dehydrogenase and alpha-hydroxycarboxylic acid 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 acidtransportergene, a catecholtransportergene and a coenzyme synthesis-related gene. The engineering bacterium has the advantages ofrealization of the efficient production of Danshensu, simple process, few impurities and great industrial application values.
A yeast having a human- or frog-origin gene encoding L-lactate dehydrogenase transferred thereinto. By producing lactic acid by using the above-described gene and a method of producing lactic acid with the use of the enzyme, it is possible to efficiently produce lactic acid which is widely usable. As a result, lactic acid can be provided at a lower cost.
The invention discloses genetic engineering compound bacteria and an application thereof in biosynthesis of PGA (phenylglyoxylic acid). The genetic engineering compound bacteria contain a novel L-lactate dehydrogenasegene LhLDH, an existing D-mandelate dehydrogenasegene and encoded genes thereof as well as a novel mandelate racemase gene HrMR, the genetic engineering compound bacteria are utilized for production and application of PGA synthesized through whole-cellcatalysis of racemic mandelic acid, the production process is simple, the conditions are mild, the efficiency is high and the cost is low. The genetic engineering compound bacteria have the benefits as follows: the constructed genetic engineering compound bacteria can realize high yield of D-mandelate dehydrogenase, L-lactatedehydrogenase and mandelate racemase; on the basis of whole-cellcatalysis, the genetic engineering compound bacteria can efficiently oxidize racemic mandelic acid for production of PGA without addition of coenzymes. Through the genetic engineering compound bacteria, the conversion rate of racemic mandelic acid can be 96% or above, the purity of PGA can be 99% or above, and the genetic engineeringcompound bacteria have good industrial prospects.
The invention provides a method for quickly distinguishing lactic acid bacterium strains. The method includes the steps that firstly, a culture solution of lactic acid bacterium strains to be detected is centrifuged, and supernate is taken and diluted by 20-25 folds; secondly, the supernate, water, a glycylglycinebuffer solution, NAD+ and D-GPT are mixed for 2-4 min before D or L-lactic dehydrogenase is added according to the table 1, a light-absorbing value A1 is read, then D-or L-LDA is added for reacting for 4-6 min, and a light-absorbing value A2 is read; thirdly, a light-absorbing value change value (A2-A1) of samples is subtracted by a contrasted light-absorbing value change value (A2-A1) to obtain deltaAD-lactic acid or deltaAL-lactic acid, the deltaAD-lactic acid or deltaAL-lactic acid is substituted into the following formula to calculate the concentration of D-lactic acid or L-lactic acid of the lactic acid bacterium strains to be detected, and please see the formula in the description. The lactic acid bacterium strains to be detected are distinguished according to the concentration ratio of D-lactic acid and L-lactic acid of the lactic acid bacterium strains to be detected.
The invention discloses an engineering bacterium, and an application thereof in the production of p-hydroxyphenyllactic acid by using a cheap substrate, and belongs to the technical field of bioengineering. The engineering bacterium constructed in the invention can simultaneously express four enzymes which are tyrosinephenollyase, L-amino acid oxidase, alpha-hydroxycarboxylic acid oxidase and L-lactate dehydrogenase respectively. The knockout or enhanced expression of a related gene on the genome of the Escherichia coli can promote the transportation of the substrate and reduce the decomposition of the product. The method for producing the p-hydroxyphenyllactic acid has the advantages of simple process, few impurities and great industrial application values.
The present invention relates to a homocysteine assay kit and its preparation method and detection method. The homocysteine assay kit includes reagent R1 and reagent R2. Reagent R1 is prepared from the following components: Aqueous solution: phosphate buffer, 1.8~2.2mL / L Triton X‑100, 0.11~0.16g / L L‑serine, 805~815KU / L lactate dehydrogenase, 0.5~0.75g / L NADH and 0.8-1.2mL / L ProcLin300; reagent R2 is an aqueous solution prepared from the following components: phosphate buffer, 1.8-2.2mL / L Triton X-100, 0.05-0.10g / EDTA dihydrate disodium salt in L, ProcLin300 in 0.8-1.2mL / L, cystathionine-β-synthase in 20.0-22.0KU / L and cystathionine-β-synthase in 10.0-14.0KU / L ‑Decomposing enzymes. The homocysteine determination kit forms a reagent R1 and a reagent R2 by screening each component containing a specific ratio, and has a low detection limit, high sensitivity, wide linear range as a whole, and good stability.