159 results about "Nicotinamide adenine dinucleotide phosphate" patented technology

The invention discloses a recombinant microorganism for preparing dharma diene and protopanoxadiol and a construction method of the recombinant microorganism. The construction method of the recombinant bacteria comprises a step of adding dharma diene synthase, protopanoxadiol synthase and nicotinamide adenine dinucleotide phosphate - cytochrome P450 reductase encoding gene into saccharomyces cerevisiae to obtain recombinant bacteria I. According to the recombinant microorganism for preparing the dharma diene and the protopanoxadiol and the construction method of the recombinant microorganism, by means of homologous recombination, the dharma diene synthase, the protopanoxadiol synthase and the nicotinamide adenine dinucleotide phosphate - the cytochrome P450 reductase encoding gene are all added into the saccharomyces cerevisiae to obtain the initial recombinant bacteria, and the effect that the initial recombinant bacteria can produce trace amount of the dharma diene and trace amount of the protopanoxadiol is discovered; tHMG1 activity of the initial recombinant bacteria is further improved, and therefore intermediate recombinant bacteria are obtained, and by means of the intermediate recombinant bacteria, the yield of the dharma diene and the yield of the protopanoxadiol are significantly increased; the activity of one or two or three of ERG1, ERG9 and ERG20 are improved on the basis of the intermediate recombinant bacteria, and the effect that the recombinant bacteria which can be used to increase the yield of the dharma diene and the yield of the protopanoxadiol are constructed is also discovered. By means of the recombinant microorganism for preparing the dharma diene and the protopanoxadiol and the construction method of the recombinant microorganism, the foundation is laid for artificially synthesizing the dammar diene and the protopanoxadiol.

The present invention provides for stable nicotinamide adenine dinucleotide (NAD / NADH) and nicotinamide adenine dinucleotide phosphate (NADP / NADPH) derivatives of formula (I), enzyme complexes of these derivatives and their use in biochemical detection methods and reagent kits.

The invention relates to a preparation method of nicotinamide adenine dinucleotide, which comprises the following steps of: with 1,2,3,5-tetraacetyl-beta-D-ribofuranose as a starting raw material, preparing nicotinamide mononucleotide through sequentially a condensation reaction, an ammonolysis reaction and a phosphorylation reaction; and then mixing the prepared nicotinamide mononucleotide with adenosine triphosphate, generating an enzymic catalytic reaction in the presence of nicotinamide adenosine nucleotide transferase and pyrophosphatase to prepare the nicotinamide adenine dinucleotide. Compared with the prior art, the technical scheme provided by the invention has the advantage that the whole process is simplified and optimized by combination of a chemical method and an enzymic method.

The present invention is related to tetrahydroindole derivatives of Formula (I), pharmaceutical composition thereof, methods of preparation thereof and to their use for the treatment and / or prophylaxis of disorders or conditions related to Nicotinamide adenine dinucleotide phosphate oxidase (NADPH Oxidase).

The present invention is related to pyrazolo pyridine derivatives of Formula (I), pharmaceutical composition thereof and to their use for the treatment and / or prophylaxis of disorders or conditions related to Nicotinamide adenine dinucleotide phosphate oxidase (NADPH Oxidase).

The present invention is related to pyrazolo pyridine derivatives of Formula (I), pharmaceutical composition thereof and to their use for the treatment and / or prophylaxis of disorders or conditions related to Nicotinamide adenine dinucleotide phosphate oxidase (NADPH Oxidase).

A bio-battery includes a biomolecular energy source, a first electrode and a second electrode. In some configurations, a bio-battery may also include a first cell containing the first electrode and the biomolecular energy source, and a second cell having a reducible substrate and the second electrode. The first cell can be in ionic communication with the second cell, for example by a proton exchange membrane. Various biomolecular energy sources can be used, including proton donor molecules or electrolytically oxidizable molecules. For example, the biomolecular energy source can be selected from the group consisting of Nicotinamide Adenine Dinucleotide (NADH), Nicotinamide Adenine Dinucleotide Phosphate (NADPH) and 5,10-Methylenetetrahydrofolate Reductase (FADH).

The present invention is related to pyrazolo pyridine derivatives of Formula (I), pharmaceutical composition thereof and to their use for the treatment and / or prophylaxis of disorders or conditions related to Nicotinamide adenine dinucleotide phosphate oxidase (NADPH Oxidase).

The present invention is related to pyrazoline dione derivatives of Formula (I), pharmaceutical composition thereof and to their use for the treatment and / or prophylaxis of disorders or conditions related to Nicotinamide adenine dinucleotide phosphate oxidase (NADPH Oxidase).

Coordinately regulated over-expression of the genes encoding glucose 6-phosphate dehydrogenase [“G6PDH”] and 6-phospho-gluconolactonase [“6PGL”] in transgenic strains of the oleaginous yeast, Yarrowia lipolytica, comprising a functional polyunsaturated fatty acid [“PUFA”] biosynthetic pathway, resulted in increased production of PUFAs and increased total lipid content in the Yarrowia cells. This is achieved by increased cellular availability of the reduced form of nicotinamide adenine dinucleotide phosphate [“NADPH”], an important reducing equivalent for reductive biosynthetic reactions, within the transgenic microorganism.

The invention relates to a method for promoting synthesis of DHA in schizochytrium limacinum grease, and particularly relates to a method for prompting synthesis of docosahexaenoic acid (DHA) in schizochytrium limacinum grease. The method is characterized in that the activity of malic enzyme in a tricarboxylic acid transport system is inhibited by adding an exogenous control factor, so that the content of the NADPH in the schizochytrium limacinum can be greatly reduced, the synthesis of a great amount of DHA can be promoted, and the exogenous control factor is sesamol. According to the method, a fermentable carbon source also can be selected to reduce metabolic flux of a pentose phosphate pathway in the bacteria, so that the content of NADPH in the schizochytrium limacinum can be greatly reduced, the synthesis of a great amount of DHA can be promoted, and the largest content of the DHA in the grease of final bacteria can reach 55.3 percent. The method is simple, easy, obvious in effect, capable of greatly increasing the content of DHA in the bacteria grease and is likely to realize the industrialized application.

The invention discloses application of sorbose reductase in preparation of (S)-4-chloro-3-hydroxyethyl butyrate by a biological method. Sorbose reductase of which the amino acid sequence is disclosed as SEQ ID NO:2 is used as a catalyst, 4-chloracetylethyl acetate is used as a substrate, and NADPH (nicotinamide adenine dinucleotide phosphate) is used a cofactor to prepare the (S)-4-chloro-3-hydroxyethyl butyrate in an asymmetric reduction mode. In the invention, the sorbose reductase of which the amino acid sequence is disclosed as SEQ ID NO:2 is applied to preparation of (S)-4-chloro-3-hydroxyethyl butyrate in an asymmetric reduction mode for the first time, which has a good effect. The enzyme activity of the sorbose reductase is up to 5.6U / mg, the yield for substrate is up to 90%, and the enantiomeric excess of the product is 100%. Besides, the invention has the advantage of high yield, and greatly lowers the production cost.

The invention discloses a gene engineering strain and an application thereof. The gene engineering strain comprises host cells and target genes transferred into the host cells, wherein the target genes include xylose reductase genes and NADPH (Nicotinamide Adenine Dinucleotide Phosphate) regenerate enzyme genes. The gene engineering strain is applied to the preparation of xylitol which is prepared through the following steps of: inoculating the engineering strain to a culture solution to culture until the OD600 is equal to 0.6-0.8, adding an inducer for inducible expression, and separating and purifying the culture solution after the inducible expression is finished. Compared with the prior art, the activity of the xylose reductase in the engineering strain is greatly improved and is 5 times as high as that of a neurospora crassa; and the gene engineering strain disclosed by the invention also contains an NADPH regeneration system, so that the xylose conversion ratio of the gene engineering strain is as high as 80.32%.

The invention discloses a method for synthesizing ursodeoxycholic acid (UDCA) and high-chiral-purity D-amino acid based on an enzyme-method coupling technology. The method comprises the following steps: putting chenodeoxycholic acid and alpha-ketonic acid into a solution system containing 7alpha-HSDH (Homoserine Dehydrogenase), DAADH and NADP (Nicotinamide Adenine Dinucleotide Phosphate) and carrying out enzyme catalysis reaction; separating a reaction solution by adopting an ultra-filtration membrane to obtain a concentrated mixed enzyme solution; regulating the pH (Potential of Hydrogen) ofa dialysis solution and crystallizing; filtering and separating to obtain 7-KLCA wet powder and filtrate; carrying out chromatographic treatment on the filtrate to obtain the D-amino acid; putting the7-KLCA wet powder into a solution system containing glucose, the NADP, the 7alpha-HSDH and GDH (Glutamate Dehydrogenase) and carrying out enzyme catalysis reaction; separating the reaction solution by adopting the ultra-filtration membrane to obtain the concentrated mixed enzyme solution; crystallizing, filtering and separating the dialysis solution, so as to obtain ursodeoxycholic acid. By adopting the method provided by the invention, UDCA and the high-chiral-purity D-amino acid can be obtained at the same time, the enzyme utilization rate is high, synthesis steps are simple and the cost isreduced; meanwhile, a metal reduction reagent and an organic solvent do not need to be added in a reaction process and conditions are mild; the method is environmentally friendly and is suitable forindustrial production.

According to the present invention, an inorganic phosphoric acid is detected by a method which includes: subjecting a sample to a measurement system containing glyceraldehyde-3-phosphate, oxidized nicotinamide adenine dinucleotide or oxidized nicotinamide adenine dinucleotide phosphate, glyceraldehyde phosphate dehydrogenase, and an electron mediator; and measuring a current value in the measurement system. In the method, a pyrophosphate is quantitatively measured with high sensitivity and at a high speed through converting the pyrophosphate in a sample into an inorganic phosphoric acid. Such a measurement of a pyrophosphate allows for quantitative determination of the pyrophosphate which is produced concurrent with the extension of a DNA, thereby enabling the detection of the presence of a targeted nucleic acid, and typing of a base in a SNP site of a targeted DNA.

The invention provides a preparation method of (R)-2-(2,5-difluorophenyl) pyrrolidine or salt thereof. N-protected pyrrolidone and a 2,5-fluorophenyl magnesium chloride Grignard reagent are subjectedto a Grignard reaction, and a compound represented as the formula 3 is obtained; the compound represented as the formula 3 is subjected to deprotection and ring closure under the acidic condition, anda compound represented as the formula 4 or salt of the compound is obtained; under enzyme catalysis, formic acid or formate is utilized as a hydrogen donor, a cofactor is adopted, cofactor cycle is realized, and the compound represented as the formula 4 or the salt of the compound is selectively reduced to synthesize an optically pure compound, namely, (R)-2-(2,5-difluorophenyl) pyrrolidine or the salt thereof; enzyme is a combination of recombinant imine reductase and formate dehydrogenase, and the cofactor is oxidized / reduced type nicotinamide adenine dinucleotide phosphate or oxidized / reduced type nicotinamide adenine dinucleotide. The method is safe in process and simple to operate, and yield and optical purity of a product are both higher.

The invention relates to a preparation method of tert-butyl (3R,5S)-6-chloro-3,5-dihydroxyhexanoate. The preparation method is characterized in that tert-butyl (S)-6-chloro-5-hydroxy-3-carbonylhexanoate is taken as a substrate and the substrate is subjected to reduction reaction in the presence of a biocatalyst, a coenzyme and coenzyme regeneration systems to generate tert-butyl (3R,5S)-6-chloro-3,5-dihydroxyhexanoate; the biocatalyst is ketoreductase; the coenzyme is NADP (nicotinamide adenine dinucleotide phosphate); the coenzyme regeneration systems are glucose and glucose dehydrogenase; the reduction reaction is carried out in a phosphate buffer solution at 25-30 DEG C in the presence of an enzyme protection reagent; the pH value of a reaction system is controlled to be 6.0-8.0 by adding a sodium carbonate solution. The preparation method has the effects of substantially increasing the concentration of the substrate and reducing the usage amount of the enzyme, so that the product has good production environment friendliness and the commercial value of industrialization is prominent.

The present invention is related to pyrazolo piperidine derivatives of Formula (I), pharmaceutical composition thereof and to their use for the treatment and / or prophylaxis of disorders or conditions related to Nicotinamide adenine dinucleotide phosphate oxidase (NADPH Oxidase).

The invention provides a novel method for synthesizing UDCA (ursodesoxycholic acid) by catalyzing CA (cholic acid) by a whole-cell combined chemical method. According to the method, cholic acid is oxidized by the aid of sodium hypochlorite to obtain dehydrocholic acid, the dehydrocholic acid is catalyzed by the aid of a built Escherichia coli engineering bacterium cell to synthesize 12-keto-ursodesoxycholic acid, and the 12-keto-ursodesoxycholic acid is reduced into the ursodesoxycholic acid by a Wolff-Kishner method. The Escherichia coli engineering bacterium cell comprises a co-expression system of three foreign genes such as 7beta-hydroxysteroid dehydrogenase (7beta-HSDH), 3alpha-hydroxysteroid dehydrogenase (3alpha-HSDH) and coenzyme NADP (nicotinamide adenine dinucleotide phosphate) +regenerative glucose dehydrogenase (GDH). The method is cheap in chemical oxidation raw material, less in catalytic by-products and by-products, high in yield and cell catalyst yield and applicable to industrial production, and addition of coenzyme is omitted.

The present invention is related to pyrazolo piperidine derivatives of Formula (I), pharmaceutical composition thereof and to their use for the treatment and / or prophylaxis of disorders or conditions related to Nicotinamide adenine dinucleotide phosphate oxidase (NADPH Oxidase).