33 results about "Formic dehydrogenase" patented technology

The invention discloses a recombinant clostridium, a method for constructing the same and application thereof. The method for constructing the recombinant clostridium is to introduce a gene for encoding formic dehydrogenase into a clostridium to construct a recombinant bacterium. The clostridium is an acetone butanol clostridium or a clostridium beijerinckii. The invention also discloses a method for producing butanol, which is to ferment and culture the recombinant bacterium constructed by the method for constructing the recombinant clostridium to produce the butanol. The yield of butanone produced through fermenting the recombinant acetone butanol clostridium constructed by the invention can reach as high as 14.1 grams per liter.

The invention relates to a method for preparing alpha-aminobutyric acid by utilizing series connection amino acid dehydrogenase and codon optimization formate dehydrogenase recombinant escherichia coli. First, codon optimization is performed on a Candida boidinii formate dehydrogenase gene sequence according to the codon preference of the escherichia coli. After codon optimization on formate dehydrogenase, the expression quantity of formate dehydrogenase in the escherichia coli can be improved remarkably, the yield of alpha-aminobutyric acid is increased, and in the escherichia coli, co-expression formate dehydrogenase and L-amino acid dehydrogenase can promote circulation of cofactors in mycetome without needing the adding of any exogenous cofactors. In addition, the process of utilizing whole-cell transformation bulk chemical L-threonine to produce alpha-aminobutyric acid is simple and quick, and the cost is low. In a 5 L fermentation tank, the yield of alpha-aminobutyric acid obtained through the method can be 81.5 g/L, and an actually effective strategy is provided for industrial production of alpha-aminobutyric acid.

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 pesticide degrading compound enzyme is prepared with lignoperoxidase, phytase, phenoloxidase, formamidase, cyanide hydrolase, formic dehydrogenase, lysozyme, and Tween solution, and through mixing, culture, drying and packing. The product has high activity, high efficiency and wide degradation spectrum, and has the functions of killing bacteria, decontaminating, eliminating pollutant heavy metal, degrading pesticide residue, etc.

The invention discloses a method for improving conversion rate of chiral amine. The method comprises the following steps: synthesizing and amplifying an optimized ArR-omega TA nucleotide sequence containing enzyme cutting sites, constructing p2A4-ArR-omega TA, p2A4-FA and p2A4-ldh, wherein FA is a gene for expressing formic acid acetyl transferase, and correspondingly introducing escherichia colito obtain recombinant bacteria 1, FA and 6; fermenting to obtain fermentation liquor of the recombinant bacteria 1, FA and 6; centrifuging to obtain fermentation liquor precipitates of the recombinantbacteria 1, FA and 6; and correspondingly preparing crude FA enzyme liquid and crude ldh enzyme liquid; resuspending the fermentation liquor precipitate of the recombinant bacteria 1 by using PBS, adding PLP and NAD< +>, and reacting; and adding the crude FA enzyme liquid, the crude ldh enzyme liquid, formate dehydrogenase, D-alanine, a substrate ketone and CoA, adding a cosolvent, reacting, terminating, extracting and drying. According to the method disclosed by the invention, a byproduct pyruvic acid is removed, and the conversion rate of the chiral amine is improved. The formate dehydrogenase is added to recover a cofactor reduced coenzyme I.

The invention discloses a method for promoting cell-catalyzed allitol production by utilizing recombinant intracellular-RNA-support-immobilized recombinase. The method comprises the steps: (1) constructing a triangular reticular RNA support for immobilizing an intracellular recombinant protein; (2) constructing recombinant plasmids for anchoring ribitol dehydrogenase and formate dehydrogenase in the triangular reticular RNA support; (3) constructing Escherichia coli engineering strains employing the triangular reticular RNA support immobilized recombinase to catalyze allitol; and (4) conducting fermentation by using the Escherichia coli engineering strains constructed in the step (3) to produce the allitol. According to the method, the ribitol dehydrogenase and the formate dehydrogenase are anchored by using the RNA support, the distance of an enzymatic reaction is shortened, and the synthesis of the allitol is efficiently promoted. Proven by experiments, the allitol yield of the engineering strains constructed by the method disclosed by the invention can be increased to 4.433g/L from 3.036g/L, so that the yield of the product is advantageously maximized in industrial practical applications.

The invention discloses a fusion protein as well as a coding gene thereof and application thereof. The fusion protein provided by the invention is protein which is obtained by in series connecting the connecting peptides of ethanol dehydrogenase and formate dehydrogenase. The ethanol dehydrogenase is from lactobacillus brevis and the formate dehydrogenase is from candida boidinii; the connecting peptides are rigid structures Linker (EAAAK). According to the experiment in the invention, the invention provides the fusion protein which is obtained by in series connecting the connecting peptides of ethanol dehydrogenase and formate dehydrogenase, wherein enzyme activity of the formate dehydrogenase is higher than the activity for independently expressing the formate dehydrogenase after the formate dehydrogenase is expressed in escherichia coli; moreover, the stability and heat stability of the ethanol dehydrogenase in organic substrate are also improved. The construction of the fusion protein provides important reference for the improvement to the preparation method for chiral alcohol and chiral hydroxy compound.

The invention discloses formic acid and CO2 autotrophic recombinant escherichia coli and a construction method thereof, and belongs to the field of microbial metabolic engineering and the field of synthetic biology. According to the invention, the most common escherichia coli is used as a starting strain, and formic acid assimilated modular plasmids and CO2 assimilated modular plasmids are introduced to construct the escherichia coli which rapidly grows by using formic acid and CO2 as a unique carbon source. The whole construction process is simple, the period is short, and the universality ishigh. According to the recombinant bacterium, the level of an intracellular reduced cofactor NAD (P) H can be improved by virtue of a formate dehydrogenase mutant, so that the recombinant bacterium has the capability of promoting pyruvic acid synthesis from beginning to end by eating formic acid, and finally formic acid and CO2 autotrophic escherichia coli is obtained; the escherichia coli is inoculated to a culture medium taking formic acid and CO2 as a unique carbon source for fermentation culture, 0.9 OD600 biomass can be obtained at most within 56 hours, and the original strain hardly grows. The escherichia coli is methanoic acid and CO2 autotrophic escherichia coli with the highest growth speed under the condition of no assistance of gene knockout, long-term domestication and other means at present.

The invention relates to a kit for diagnosing/measuring ammonia (ions) by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of ammonia (ions), and belongs to the technical field of medical/food/environmental inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, glutamic acid, adenyl pyrophosphate, oxaloacetic acid, glutamine synthetase, phosphoric acid enol pyruvate carboxylase, formic dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet/visible light analyzer, and the degree/velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the concentration of ammonia (ions).

The invention relates to a kit for diagnosing/measuring kalium (ions) by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of kalium (ions), and belongs to the technical field of medical/food/environmental inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, adenosine diphosphate, enolphosphopyruvate, ferricytochrome b1, pyruvate kinase, pyruvate dehydrogenase, formic dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet/visible light analyzer, and the velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the concentration of kalium (ions).

The invention relates to a kit for diagnosing/measuring glycine by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of the glycine, and belongs to the technical field of medical/food inspection and measurement. The main components of the kit include a buffer solution, coenzyme, methyl sulfate phenazine, ferricytochrome B1, hydrogen cyanide synthetase, formic dehydrogenase, NAD dependent formic dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet/visible light analyzer, and the degree/velocity of the increase in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the active concentration of the glycine.

The invention relates to a kit for diagnosing/measuring potassium (ions) by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of the potassium (ions), and belongs to the technical field of medical/food/environmental inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, adenosine diphosphate, a phosphoenolpyruvic acid, coenzyme A, pyruvate kinase, pyruvate oxidase, formic dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet/visible light analyzer, and the velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the concentration of potassium (ions).