40 results about "Decoyinine" patented technology

The invention provides a mutant of 7 beta-hydroxyl steroid dehydrogenase, application of the mutant and a synthesis method. The mutant of the 7 beta-hydroxyl steroid dehydrogenase is characterized in that amino acid sequences of the mutant are Seq ID NO:4, and coded nucleotide sequences are Seq ID NO:3; or amino acid sequences of the mutant are Seq ID NO:6, and coded nucleotide sequences are Seq ID NO:5. The mutant, the application and the synthesis method have the advantages that cholic acid compounds, particularly ursodeoxycholic acid, can be catalytically synthesized by the efficient 7 beta-hydroxyl steroid dehydrogenase, mutant enzymes of the 7 beta-hydroxyl steroid dehydrogenase and coenzyme regeneration systems, accordingly, the substrate concentration can reach 100 g/L, the conversion rate is 99.2-99.5%, and the weight yield can reach 94-96%; and the enzymes can be inexpensively and easily obtained by the aid of a fermentation process, accordingly, the production cost and the product quality are superior to the production cost and the product quality of chemical synthesis methods, and the mutant and the synthesis method are applicable to industrial production.

The invention discloses a method for producing acetoin by efficient bioconversion of 2,3-butanediol by using a Bacillus subtilis nicotinamide adenine dinucleotide (NAD)<+> regeneration system, and belongs to the field of genetic engineering. Acetoin reductase and educed form of nicotinamide-adenine dinucleotide (NADH) oxidase in a high-yield acetoin strain B.subtilis JNA with independent intellectual property rights, which are autonomously screened from a laboratory are cloned by the method disclosed by the invention, and excessive coexpression of the acetoin reductase and NADH oxidase in B.subtilis JNA is carried out, so that the production of acetoin by effective conversion of the 2,3-butanediol in the wild-type high-yield acetoin bacillus subtilis by virtue of the NAD<+> regeneration system is realized for the first time at home and abroad. The enzyme activity determination and intracellular coenzyme level research on the built gene engineering strain prove that the 2,3-butanediol can be lastingly and effectively converted by B.subtilis JNA/pMA5-bdhA-yodC to produce the acetoin. 120g/L of 2,3-butanediol can be finally converted into about 92.5g/L of acetoin by the B.subtilis JNA/pMA5-bdhA-yodC when the temperature is 40 DEG C and the pH is 8.0 under the optimal whole-cell conversion condition of adding 5mM of MnC12, the acetoin yield can be up to 2.31g/(L.h), and is the highest lever for producing the acetoin from the bacillus subtilis in the current report, and a foundation is provided for industrial production of the acetoin from microorganisms.

The invention discloses a construction method of a metabolic engineering escherichia coli strain for producing hydracrylic acid from acetic acid. Escherichia coli reformed by metabolic engineering isutilized for producing hydracrylic acid by fermenting acetic acid; and construction ways are as follows: constructing a metabolic way for producing hydracrylic acid from acetyl CoA, and/or over-expressing relevant genes of an acetic acid intake way so as to increase the transmission rate of acetic acid, and/or blocking or decreasing TCA circle so as to increase metabolic flow of acetyl CoA flowingtowards a target metabolite, and/or reducing decarboxylic reaction between malic acid or oxaloacetic acid so as to delete a byproduct generation way, and/or deleting key genes in an alcohol production way so as to regulate metabolic flow of an acetyl CoA node and/or regulating intracellular oxidation reduction balance through coenzyme engineering. According to the construction method, a production way for exogenously expressing hydracrylic acid is constructed, the metabolic ways, the adjustment and control are analyzed, and escherichia coli is reformed by virtue of a metabolic engineering measure, so that the prepared strain can be utilized for producing hydracrylic acid in a culture medium taking acetic acid as a carbon source.

The invention discloses a lysine decarboxylase strain and an application method thereof in the production of pentamethylenediamine. The lysine decarboxylase strain adopts recombinant bioengineering bacteria obtained by introducing a lysine decarboxylase gene into escherichia coli by a genetic engineering method as a starting strain, a high enzyme activity lysine decarboxylase strain is obtained bymutagenesis screening, the preservation number is CGMCC No. 16762, and compared with original engineering bacteria, the lysine decarboxylase activity is increased by 3.93 times. The invention furtherdiscloses the application of the lysine decarboxylase strain in the production of the pentamethylenediamine, the lysine decarboxylase strain is subjected to fermentation culture to obtain a fermentation broth containing the lysine decarboxylase, the fermentation broth is added in an amount of 5% in the lysine hydrochloride and a coenzyme solution, the conversion is carried out for 3 h at 40 DEG C, the highest content of the pentanediamine is 326.3 g / L, and the molar conversion of the lysine hydrochloride reaches 99.98%.

The invention discloses a genetically engineered bacterium, and applications thereof in preparation of L-phosphinothricin. The genetically engineered bacterium comprises Escherichia coli and recombinant plasmids transferred into Escherichia coli; the recombinant plasmids contain transaminase gene and pyridoxal phosphate synthetic route enzyme gene; the nucleotide sequence of the transaminase geneis represented by SEQ ID NO.1; the nucleotide sequence of the pyridoxal phosphate synthetic route enzyme gene is represented by SEQ ID NO.2-5. According to the applications, genetic engineering technology is adopted to express ribose-5-phosphate pathway synthesis key gene PdxST in Escherichia coli, or enhance Escherichia coli self PLP synthesis pathway key gene epd, pdxJ, and dxs expression; preparation of the coenzyme reinforced transaminase genetically engineered bacterium through construction is capable of increasing intracellular coenzyme PLP content obviously, avoiding adding of externally-sourced PLP in production of L-phosphinothricin using the genetically engineered bacterium, increasing transaminase enzyme activity and stability obviously, prolonging transaminase half life, reducing production cost, and increasing L-phosphinothricin production efficiency.

The present invention discloses genetically engineered bacteria and application thereof in production of coenzyme Q10. The genetically engineered bacteria comprises coenzyme Q10 producing bacteria and desired genes transferred into the coenzyme Q10 producing bacteria, and the desired genes are a demethylnaphthoquinone methyltransferase gene, a 2-octo isoprene-3-methyl-6-methoxy-1,4-hydroquinone hydroxylase gene and a 3-demethyl ubiquinone-93-methyl transferase gene. Rate-limiting enzyme genes UbiE, UbiG and UbiF in a coenzyme Q10 synthetic route are transferred into rhodobacter sphaeroides, and by in-series over-expression of the three rate-limiting enzyme genes UbiE, UbiG and UbiF, coenzyme Q10 synthesizing capability of the rate-limiting enzyme genes can be strengthened; and compared with the rhodobacter sphaeroides with no desired gene being transformed, by use of the genetically engineered bacteria for coenzyme Q10 producing, the yield of the coenzyme Q10 is increased by more than 20%.

The present invention discloses construction and applications of nicotinamide adenine dinucleotide (NAD) auxotrophic Escherichia microbe, wherein NAD transporter protein is introduced to the Escherichia microbe, an intracellular NAD biosynthesis pathway is blocked, and the Escherichia engineered bacteria growing dependent on exogenous pyridine nucleotide coenzyme is constructed. The NAD auxotrophic Escherichia can be used in the field of bio-technology, and can be provided for screening inhibition drugs for pyridine nucleotide coenzyme metabolism, improving oxidation reduction biotransformation efficiency, and establishing non-antibiotic screening platforms for gene cloning and protein heterogenesis expression.

The invention discloses a method for producing coenzyme Q10 by fermenting genetic recombinant bacteria. According to the method, an NAD (Nicotinamide Adenine Dinucleotide) kinase gene is transplanted into a strain of efficient expression reducing coenzyme Q10. The culture conditions of recombinant bacteria optimization are inclined activation, seed culture and fermental cultivation. According to the method disclosed by the invention, the recombinant bacteria constructed by genetic engineering means are strong in stability; the yield of the coenzyme Q10 of the recombinant bacteria is 1.63g/L and increased by 21.6 percent compared with an original strain; the coenzyme Q10 can meet the requirement on industrial production.

The invention provides a method for catalytic synthesis of a sitagliptin chiral intermediate (3R)-3-amino-1-[3-(trifluoromethyl)-5,6,7,8-teralin-1,2,4-triazol[4,3-a]pyrazine-1-yl]-4-(2,4,5-trifluoro-phenyl)buta-1-one by means of engineering bacterium cells containing transaminase-coenzyme pyridoxal phosphate co-immobilization Escherichia coli, E.coli). The method comprises the steps of culturing engineering bacteria containing transaminase Escherichia coli, preparing co-immobilization cells, and synthesizing the immobilization cell asymmetric catalysis itagliptin chiral intermediate. The transaminase-PLP co-immobilization cells are adopted as a catalyst, the stability is good, the service life is long, the organic solvent is good in tolerance, repeated usage can be performed, no expensiveexogenous coenzyme needs to be added in the reaction process, and the production cost is greatly lowered. The method is simple in technology, low in cost and high in product yield and purity, and hasthe quite high application value in the industrial production of the sitagliptin chiral intermediate.

A method for the identification and treatment of pathogenic microorganism infections by inhibiting one or more enzymes in a metabolic pathway by inhibiting the conversion of substrate to produce the penultimate or ultimate product particularly by inhibiting the activity of one or more of the enzymes in the pathway, and compounds and pharmaceutical compositions for inhibiting infections of pathogenic microorganisms by inhibiting such enzymes.

The invention relates to an escherichia coli coenzyme metabolic-pathway transformation and biotransformation method. According to the method, a coenzyme metabolic pathway is transformed by a homologous recombination method, a coenzyme degradation related gene yjaDyrfE in protein expression host bacterium escherichia coli JM109 (DE3) is removed and the concentration of endogenous coenzyme during reproduction of the host bacterium is increased. When a host bacterium expression enzyme system transformed by the method is used for preparing chiral molecules by biotransformation by a biological reduction method, the addition of coenzyme in a transformation system can be remarkably reduced or no coenzyme is added in the transformation system.

The invention discloses a genetic recombinant engineering bacterium and application thereof in catalytic synthesis of D-pantolactone and belongs to the field of biocatalytic research. The genetic recombinant engineering bacterium takes escherichia coli as a host bacterium, the host bacterium contains recombinant expression plasmids containing a conjugated polyketide reductase gene and a glucose dehydrogenase gene, and the nucleotide sequence of the conjugated polyketide reductase gene is shown as SEQ ID NO.1. The engineering bacterium simultaneously expresses polyketide reductase and glucose dehydrogenase through induction; the engineering bacterium is applied to the asymmetric catalytic synthesis of D-pantolactone, the in-situ regeneration of a coenzyme NADPH is realized through a synergistic effect of the two enzymes, and the purpose of efficient production from keto-pantolactone to D-pantolactone can be achieved. A method simplifies a production process, avoids addition of extra coenzymes at the same time and reduces the production cost.

The invention relates to a kind of double enlarge method, enzyme colorimetric method and enzyme linked method and a diagnosis reagent box applying the malic acid dehydrogenase coupling pyruvate dehydrogenase enzymatic reaction ratio colorimetric method/end-point method. The reaction of the malic acid dehydrogenase enzymolysis malic acid generates pyruvate and at the same time makes the coenzyme reducing the reduction coenzyme. The pyruvate generated by acid dehydrogenase enzymolysis malic acid makes the coenzyme reducing the reduction coenzyme again by the coupling reaction of pyruvate dehydrogenase. Because of making the coenzyme reducing the reduction coenzyme two times the reaction sensitivity is enhanced double. So we can assay the degree/speed of the ascent absorbance at the place of 340nm. It can measure and calculate the density of citric acid by measuring the degree/speed of the fall-way absorbance at the place of 340nm. The method has high specificity and it is not polluted by the endogenous and exogenous object and the test result is precise and accurate. The invention can get the array result by the ultraviolet/visible light analytic instrument so it is convenience to extend and apply.

The invention relates to a monoamine oxidase diagnosis reagent box of Enzymatic Recycling Method, and the invention also relates to method principle for detecting the solution of monoamine oxidase, constitute and component of reagent, which belongs to the technical field of medical checking measurements. The reagent box in the invention can be dry powder state, and used after dissolution; it also can be formulated to be liquid agent for direct usage. The component of reagent box mainly contains: buffering liquid, alpha- ketoglutaric acid, glutamate dehydrogenase EC 1.4.1.2, EC 1.4.1.3, EC 1.4.1.4, Glutamate oxidase EC 1.4.3.7, EC 1.4.3.11, reduction type coenzyme and stabilizer, and the component can be matched to form single-reagent reagent box, two-reagent reagent box, and three- reagent box; The enzymatic reaction is generated between sample and reagent by mixing sample and reagent with definite volume, then the reactants is placed under the ultraviolet/visible light analyzer to detect ascend degree of absorbance at main wavelength of 340nm, so concentration of monoamine oxidase can be calculated. The invention can obtain the detecting result by ultraviolet/visible light analyzer, and the sensibility is high, precision is well, and easy to be spread.