8 results about "Formate dehydrogenase H" patented technology

This invention discloses an enzyme preparation for catalytic synthesis of epiinositol and a method for synthesizing epiinositol, belonging to the field of genetic engineering technology. The enzyme preparation includes squalinositol dehydrogenase, NADH oxidase, formate dehydrogenase, and muscle inositol dehydrogenase; the amino acid sequence of squalinositol dehydrogenase is shown in SEQ ID NO. 6, and the amino acid sequence of muscle inositol dehydrogenase is shown in SEQ ID NO. 8. The enzyme activities of the squalinositol dehydrogenase mutant and the muscle inositol dehydrogenase mutant obtained by mutation in this invention are significantly increased, thus further improving the yield and conversion rate of epiinositol during catalytic synthesis. Furthermore, the enzyme preparation of this invention also contains NADH oxidase and formate dehydrogenase, achieving in-situ regeneration of NADH, thereby effectively reducing the amount of cofactors and NADH used in the catalytic synthesis of epiinositol, and thus reducing production costs.

The invention discloses short-chain dehydrogenase for synthesis of (S)-1-(2-methoxy-3-bromophenyl) ethanol as well as a mutant, a coding gene, a plasmid, a genetically engineered bacterium and application of the short-chain dehydrogenase, and belongs to the technical field of biocatalysts. The mutant is obtained by mutating methionine at the 97th site into isoleucine on the basis of a wild type enzyme shown in SEQ ID NO.1 (SEQ ID NO.3). The invention further discloses a preparation method of the short-chain dehydrogenase. The invention also provides a coding gene, an expression vector, a genetically engineered bacterium and a double-enzyme catalytic system containing the mutant and formate dehydrogenase. The system can efficiently and highly selectively catalyze and synthesize the target chiral alcohol under mild conditions, and the conversion rate is gt; 95%, eet; 99% of the method is suitable for green preparation of the key intermediate of the medicine lutrombopag.

This invention discloses a method for preparing 1,2-pentanediol, comprising using 2-pentanone as a substrate, adding 2-keto reductase, linalool dehydratase, formate dehydrogenase, lysozyme, coenzyme NAD+, and water to react and obtain n-pentene. The obtained n-pentene is then thoroughly mixed with ethanol. This mixture is then fed into a reaction system containing olefin monooxygenase, epoxide hydrolase, formate dehydrogenase, lysozyme, coenzyme NAD+, MgSO4, and water using a fed-batch method. After the n-pentene and ethanol mixture has been completely added, the reaction continues for a period of time, followed by extraction to obtain 1,2-pentanediol. This preparation method uses 2-pentanone as a raw material, catalyzing the synthesis of 2-pentanol via 2-keto reductase. 2-pentanol is then catalyzed by a dehydrating enzyme to generate n-pentene, which is then catalyzed by an olefin monooxygenase to generate 1,2-epoxypentane. Finally, epoxide hydrolase ring-opens the 1,2-pentanediol. This process requires only four enzymatic catalytic reactions for efficient synthesis.

The present invention relates to a genetically engineered microorganism expressing (i) formate tetrahydrofolate (THF) ligase, methenyi-THF cyclohydrolase and methylene-THF dehydrogenase, (ii) the enzymes of the glycine cleavage system (GCS), (iii) serine deaminase and serine hydroxymethyltransferase (SHMT), (iv) an enzyme increasing the availability of NADPH, and (v) optionally formate dehydrogenase (FDH), and wherein the genetically engineered microorganism has been genetically engineered to express at least one of the enzymes of (i) to (v), wheren said enzyme is not expressed by the corresponding microorganism that has been used to prepare the genetically engineered microorganism, and wherein the enzymes of (i) to (v) are genomically expressed.

Provided in the present invention are a formate dehydrogenase mutant and use thereof. The formate dehydrogenase mutant comprises: (a) a protein having an amino acid sequence set forth in SEQ ID NO: 1; or (b) a protein that has been subjected to amino acid mutation at at least one of the following sites of the amino acid sequence in (a): A10, C23, K47, E53, I103, K109, V120, H148, V152, A168, L184, N187, E202, F285, Q287, T321, or K328, and that possesses formate dehydrogenase activity; and (c) a protein having no less than 80% homology with the amino acid sequence defined in any one of (a) and (b) and having a formate dehydrogenase function. The formate dehydrogenase mutant of the present application exhibits good tolerance and high activity under extreme conditions and demonstrates good suitability for industrial scale-up with low costs and high yield.

The present application belongs to the technical field of hydrogen conversion and energy storage, and particularly relates to a method for expressing multi-subunit hydrogenase and producing formic acid by using formate dehydrogenase. The present application uses a pBBR1MCS-1 vector, adds a Lac promoter in front of each subunit sequence of hydrogenase, so that each gene is controlled by a single promoter, and adds a His tag to each gene for subsequent purification of expressed protein. After that, the constructed vector is transformed into Ralstonia eutropha H16 competent cells to obtain an engineering bacterium and express hydrogenase in the engineering bacterium. The recombinantly expressed hydrogenase is combined with formate dehydrogenase to realize self-circulation regeneration of NAD + / NADH in the reaction system without additional NADH, thereby efficiently converting H2 and CO2 into formic acid in a green and economical manner, and realizing resource utilization of green hydrogen and high-value conversion of greenhouse gas CO2.

The invention belongs to the technical field of enzyme catalysis, and particularly relates to formate dehydrogenase and application of a mutant thereof in carbon dioxide reduction. According to the present invention, the ArFDH derived from the Ancylobacter rudongensis is obtained through the gene mining, and the ArFDH derived from the Ancylobacter rudongensis is obtained; the purified protein of the enzyme is successfully obtained through the steps of codon optimization, gene synthesis, gene cloning, expression purification and the like. The enzyme activity of the recombinant ArFDH reaches 46.06 U / g, which is 1.55 times higher than that of the template enzyme TsFDH with the highest activity reported. Compared with a wild type ArFDH, the mutant I123F / D222M has the advantages that the catalytic efficiency on HCO3 <-> / CO2 is improved by 6.69 times, the substrate catalytic efficiency of enzyme is integrally improved, and the substrate specificity is enhanced. The invention not only provides a new way for catalyzing and utilizing CO2 through a biological enzyme method, but also lays a foundation stone for constructing sustainable artificial carbon cycle.

Provided are an NADP-type formate dehydrogenase mutant and the use thereof in an enzyme-catalyzed redox reaction. The mutant takes NADP-type formate dehydrogenase XSFDH derived from Xanthobacter sp. 91 with an amino acid sequence as shown in SEQ ID NO: 2 as an original sequence, and has an amino acid sequence with corresponding mutation sites (such as D222Q). Further provided are a coding gene of the NADP-type formate dehydrogenase mutant, a vector, a bioengineered bacterium, and a method for synthesizing Pro-Xylane. The mutant has the advantages of good stereoselectivity, high optical purity of the product, and an improved reaction yield.