33 results about "Formic dehydrogenase" patented technology

The invention discloses a method for preparing highly pure L-tertiary leucine through a biosynthesis process. The method comprises the following steps: inoculating an Escherichia coli seed liquid expressing leucine dehydrogenase gene and formic dehydrogenase gene into in a fermenting culture medium of a self-induction system, carrying out fermenting culture, centrifuging to obtain crude thalli, and adding a buffer solution to obtain a cell suspension; adding substrates comprising trimethylpyruvic acid and ammonium formate, and carrying out a biological catalysis reaction to obtain an trimethylpyruvic acid conversion solution; and centrifuging the conversion solution, adjusting the pH value of the solution, carrying out column chromatography, precipitating, and concentrating to obtain highly pure L-tertiary leucine. The method fully uses the high effectiveness, the specificity and the mildness of an enzyme; and compared with traditional chemical synthesis methods, the method disclosed in the invention has the advantages of high selectivity, mild conditions, simple process, low cost, small pollution and the like.

The invention provides an efficient hydrogen-production functional gene vector pET32a-fdhF as well as construction and application thereof and relates to the technical field of genetic engineering. The gene vector pET32a-fdhF is a pET32a plasmid containing formate dehydrogenase fdhF genes of Bacillus cereus. The construction of the gene vector comprises the steps of amplifying segments of the formate dehydrogenase fdhF genes in Bacillus cereus by virtue of a design primer, linking the segments to a pET32a segment containing a corresponding linking terminal by virtue of recombinase, carrying out gene sequencing, and verifying the integrity of the genes. The pET32a-fdhF plasmid is converted into bacteria, and the bacteria are cultured in a hydrogen-production culture medium, so as to produce hydrogen. The pET32a-fdhF can be applied to different bacteria, so that the bacteria with no hydrogen-production capacity can produce hydrogen, the hydrogen-production of the bacteria capable of producing hydrogen is obviously improved, and more hydrogen can be rapidly produced by virtue of the same substrate concentration. According to the pET32a-fdhF, existing escherichia coli for industrial hydrogen production can be improved.

The invention discloses a single-cell factory for efficiently synthesizing L-phenylglycine and its construction and application, belonging to the technical field of microorganisms. The present invention first realizes the high-efficiency expression of leucine dehydrogenase derived from Bacillus cereus in Escherichia coli, and obtains mutant N71S with significantly improved reduction properties through site-directed mutation, and combines the mutant enzyme with formate dehydrogenase mutant Co-expressed in Escherichia coli to form an intracellular in situ cofactor NADH cycle system, optimized the expression of formate dehydrogenase mutants through promoter and RBS sequence optimization, successfully constructed a recombinant Escherichia coli single-cell factory, and used the single-cell factory Carry out whole-cell transformation to prepare L-phenylglycine. The conversion process of this method is simple and fast, with low cost and no by-products, which is beneficial to separation and purification. After 4 hours of conversion in a 5L fermenter, the output of L-phenylglycine can reach 105.7g / l. The conversion rate was 93.3%, and the space-time yield of L-phenylosine was 26.3 g / L·h, which provided a practical and effective strategy for the industrial production of L-phenylosine.

The invention provides a method for asymmetrically synthesizing (2S, 3R)-p-thiamphenicol phenylserine in whole cells with "one bacterium and multiple enzymes", which belongs to the field of biocatalysis and enzyme engineering. By simultaneously expressing L-threonine transaldolase (PsLTTA), alcohol dehydrogenase (ApADH) and formate dehydrogenase (CbFDH) in E. Asymmetric and efficient biosynthesis of sulfonylphenylserine. The present invention can obtain the key chiral building block (2S, 3R)-p-thiamphenicol-phenylserine of β-aminoalcohol antibiotics through one-step reaction under normal temperature and pressure, and at the same time remove the inhibitory effect of by-product acetaldehyde on PsLTTA , is an efficient green biosynthetic pathway.

The invention relates to promotion of synthesis of (R)-styrene glycol by coupling (R)-carbonyl reductase and formic dehydrogenase, which belongs to the technical field of asymmetrical transformation of biological catalysis. The invention provides a recombinant Escherichia coli, namely E.coli Rosetta / pETDuet-rcr-fdh, with a preservation number of CCTCC NO: M208123, wherein the (R)-special carbonyl reductasei and the formic dehydrogenase are coupled by a co-expression mode; asymmetrical transformation reaction is catalyzed by utilizing a cultured recombinant strain and taking 2-hydroxyacetophenone as a substrate; and the optical purity of the (R)-styrene glycol reaches 100 percent e.e., and the yield of the (R)-styrene glycol reaches 85.9 percent by optimizing the biotransformation reaction conditions. The invention utilizes double-enzyme coupling technology to solve the problem of limitation of regeneration cycle of coenzyme under the condition of high-concentration substrate, provides an effective path for high-efficiency and low-cost preparation of the (R)-styrene glycol, and lays a foundation for industrial application of the biosynthesized (R)-styrene glycol.