228 results about "Alcohol dehydrogenase (NADP+)" patented technology

The invention relates to the genetic modification of Cyanobacteria for the production of ethanol, and more particularly, to the genetic modification of Cyanobacteria by incorporating the genetic information encoding for pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adh).

The present invention relates to recombinant microorganisms comprising biosynthetic pathways and methods of using said recombinant microorganisms to produce various beneficial metabolites. In various aspects of the invention, the recombinant microorganisms may further comprise one or more modifications resulting in the reduction or elimination of 3 keto-acid (e.g., acetolactate and 2-aceto-2-hydroxybutyrate) and / or aldehyde-derived by-products. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the Saccharomyces clade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms.

The invention discloses application of alcohol dehydrogenase with amino acid sequence disclosed as SEQ ID NO:2 in preparing ethyl (R)-4-chloro-3-hydroxy butyrate from ethyl 4-chloroacetoacetate by asymmetric reduction. By using alcohol dehydrogenase with amino acid sequence disclosed as SEQ ID NO:2 as a catalyst, ethyl 4-chloroacetoacetate as a substrate and NADH (nicotinamide adenine dinucleotide) as a cofactor, asymmetric reduction is carried out to prepare the ethyl (R)-4-chloro-3-hydroxy butyrate. The invention applies the alcohol dehydrogenase with amino acid sequence disclosed as SEQ ID NO:2 in preparing ethyl (R)-4-chloro-3-hydroxy butyrate from ethyl 4-chloroacetoacetate by asymmetric reduction for the first time, and has favorable effect. The enzyme activity is up to 5.6 U / mg, the yield of the substrate is up to 94%, and the enantiomeric excess value of the product is 100%. The yield is high, and the production cost is greatly lowered.

The invention discloses an alcohol dehydrogenase, a gene thereof, a recombinant expression vector and a recombinant expression transformant respectively containing the gene, a recombinase of the alcohol dehydrogenase, and an application of the alcohol dehydrogenase in asymmetric reduction synthesis of chiral diaryl secondary alcohol as a catalyst, and belongs to the technical field of bioengineering. The alcohol dehydrogenase is from Kluyveromyces sp. CCTCCM2011385, has a carbonyl group reduction function, and also has a hydroxy group oxidation function. Extra addition of glucose dehydrogenase and other enzymes used for cofactor circulation is not needed when the alcohol dehydrogenase is used in the reduction of diaryl ketone into the chiral diaryl secondary alcohol as a biocatalyst, and the alcohol dehydrogenase has the advantages of high catalysis efficiency, mild reaction conditions, easy product recovery and low cost, so the alcohol dehydrogenase has very good application and exploitation prospect in the production of antihistamine medicines.

Bacterial polynucleotides and polypeptides are provided in which the polypeptides have a dehydrogenase activity, such as an alcohol dehydrogenase (ADH) activity, an uronate, a 4-deoxy-L-erythro-5-hexoseulose uronate (DEHU) ((4S,5S)-4,5 dihydroxy-2,6-dioxohexanoate) hydrogenase activity, a 2-keto-3-deoxy-D-gluconate dehydrogenase activity, a D-mannuronate hydrogenase activity, and / or a D-mannnonate dehydrogenase activity. Methods, enzymes, recombinant microorganism, and microbial systems are also provided for converting polysaccharides, such as those derived from biomass, into suitable monosaccharides or oligosaccharides, as well as for converting suitable monosaccharides or oligosaccharides into commodity chemicals, such as biofuels. Commodity chemicals produced by the methods described herein are also provided.

The invention provides a method for producing 1,3-propylene glycol through fermentation via recombinant microbes. According to the invention, an endogenous dihydroxypropanone phosphatein phosphatase gene hdpA is over-expressed in Corynebacterium glutamicum to reinforce removal of phosphoric acid from dihydroxypropanone phosphatein so as to produce dihydroxypropanone; exogenous glycerol dehydrogenase is introduced to convert dihydroxypropanone into glycerin; and glycerin finally produces 1,3-propylene glycol under the action of exogenous glycerol dehydratase and an activator thereof and alcohol dehydrogenase. Corynebacterium glutamicum can use different cheap raw materials for fermentation, and cheap corn steep liquor can be used as a nutritional component to replace expensive yeast powder, so cost for raw materials is further reduced, and the problems in biosecurity and tolerance of the bacterial strain to a substrate and a product are overcome; and thalli obtained in the process of fermentation can be used as a product for a feed additive. The method provided by the invention produces few by-products and can further simplify the separating process of 1,3-propylene glycol.

The invention provides, inter alia, methods for the production of acetone, isopropanol and / or precursors of acetone and / or isopropanol by microbial fermentation of substrates comprising CO, genetically modified microorganisms of use in such methods, nucleic acids suitable for preparation of genetically modified microorganisms, a novel alcohol dehydrogenase and nucleic acids encoding same.

The present invention relates to a multi-stage process for producing substituted, optically active alcohols, comprising an enzyme-catalyzed synthesis step, in particular a synthesis step which is catalyzed by an alcohol dehydrogenase. The inventive method is particularly suitable for producing phenylephrine, i.e. 3-[(1R)-1-hydroxy-2-methylamino-ethyl]-phenol.

The invention discloses escherichia coli gene engineering bacteria for four enzyme co-expression. The engineering bacteria is characterized by introducing an L-amino acid oxidase gene, an alpha-ketonic acid decarboxylase gene, an alcohol dehydrogenase gene, and an enzyme gene capable of reducing NAD(P) to NAD(P)H. The invention further discloses a construction method and application of recombinantescherichia coli. The engineering bacteria is applied to biological synthesis of phenylethyl alcohol compounds, has the characteristics of simple operation, low cost, high product synthetic efficiency, and high optical purity, and has bright industrial prospects.

The invention discloses a protein and amorphous metal organic framework compound and a preparation method thereof. The organic framework compound has a 2 nm-50 nm mesoporous structure. The preparationmethod comprises the following step of enabling a protein, zinc ions and an organic ligand to react in a solvent, wherein the organic ligand is a compound containing an imidazole group; the protein is one or a combination of several kinds of cytochrome C, cytochrome P450, horse radish peroxidase, alcohol dehydrogenase, lipase, acetylcholin esterase, laccase, a green fluorescent protein, glucose dehydrogenase, glucose oxidase, trypsin, bacillus subtilis protease, carbonic anhydrase, aldehyde ketone reductase, amylase, saccharase, superoxide dismutase, urease and catalase. The preparation method of the protein and amorphous metal organic framework compound provided by the invention is simple to operate and mild in condition, the obtained product is high in protein embedding rate and good inprotein stability, and the biologic al activity of the protein is reserved to a greater extent.

The present invention provides isolated nucleic acid molecules which encode a mutant acetaldehyde-CoA / alcohol dehydrogenase or mutant alcohol dehydrogenase and confer enhanced tolerance to ethanol. The invention also provides related expression vectors, genetically engineered microorganisms having enhanced tolerance to ethanol, as well as methods of making and using such genetically modified microorganisms for production of biofuels based on fermentation of biomass materials.

The invention discloses a gene combination, a primer and a probe for detecting alcoholic liver disease susceptibility and application. The gene combination comprises a combination of three genes closely related with an alcoholic liver disease, namely an ADH2 gene of alcohol dehydrogenase, an ALDH2 gene of acetaldehyde dehydrogenase and a CYP2E1 gene of cytochrome P4502. The gene combination comprises the following SNP sites: rs1229984 site of the ADH2 gene, rs671 site of the ALDH2 gene and rs2031920 site of the CYP2E1 gene. Whether the detected crowds carry 'alcoholic liver disease susceptible genes' are comprehensively detected and analyzed by detecting a group of genes and sites related with the alcoholic liver disease susceptibility, using the specific primer and the probe and combining a mononucleotide extending technique and a micro array chip technique so as to screen the alcoholic liver disease susceptible crowd from the crowds, change unhealthy lifestyles and fulfill the purpose of preventing.

The invention discloses a method for synthesizing chiral 2-amino-1-butanol. The method comprises the following steps: 1,2-butanediol is used as a substrate and is catalyzed by enzyme A and coenzyme thereof to generate 2-ketone-1-butanol; the 2-ketone-1-butanol serves as a substrate, and the 2-amino-1-butanol is generated through catalytic reaction of enzyme B and coenzyme thereof. The enzyme A isselected from alcohol dehydrogenase, carbonyl reductase or mutants of the two enzymes; the enzyme B is selected from amino acid dehydrogenases, transaminases or mutants of the two enzymes. The invention provides a brand-new green biosynthesis route. The cheap 1,2-butanediol is used as a raw material, and the chiral 2-amino-1-butanol is synthesized through multi-enzyme co-expression or cascade or step-by-step catalysis, namely (S)-2-amino-1-butanol and (R)-2-amino-1-butanol.

The invention discloses an enzyme combination for producing L-phosphinothricin. The enzyme combination comprises glutamate dehydrogenase and a coenzyme regenerating enzyme, wherein the coenzyme regenerating enzyme is alcohol dehydrogenase, formate dehydrogenase and phosphite dehydrogenase. The invention also discloses a production method of L-phosphinothricin, 4-(methylhydroxyphosphinyl)-2-oxobutyric acid is used as a raw material, NH<4><+>, a coenzyme NADP<+> / NADP, and a coenzyme regeneration substrate are added, and then the enzyme combination is used for catalysis, wherein the glutamate dehydrogenase is used to catalyze a reaction of 4-(methylhydroxyphosphinyl)-2-oxobutyric acid to obtain L-phosphinothricin, and the coenzyme regenerating enzyme is used to reduce NADP<+> to NADP. According to the enzyme combination and the production method of L-phosphinothricin provided by the invention, by-products produced are very easy to remove, a post-treatment process of the product is simplified, the total yield of the product is greater than 95%, and the production cost of L-phosphinothricin is reduced, so that the method is a green, environment-friendly, and low-carbon process route, and is suitable for large-scale industrial production applications.

The invention relates to a preparing method for a gelatin microsphere of fixed alcohol dehydrogenase by micro-porous membrane permeation and emulsification. The method comprises the following steps that solid gelatin and alcohol dehydrogenase are dissolved in deionized water, and a gelatin / alcohol dehydrogenase solution called as a water phase is prepared; oil soluble emulsifier span-80 is added into liquid paraffin, and liquid paraffin oil called as an oil phase is prepared; a micro-porous membrane is immersed into the liquid paraffin oil, according to the volume ratio of the gelatin / alcohol dehydrogenase solution and the liquid paraffin oil, the gelatin / alcohol dehydrogenase solution is pressed to penetrate through the micro-porous membrane and enter into the liquid paraffin oil, and W-shaped or O-shaped emulsion is formed by the emulsification; glutaraldehyde is added into the W-shaped or O-shaped emulsion, and after cross linking, solidifying, washing and drying are carried out, the gelatin microsphere of the fixed alcohol dehydrogenase is obtained. The preparing method for the gelatin microsphere of the fixed alcohol dehydrogenase by the micro-porous membrane permeation and emulsification has the advantages that the preparing condition is warm, the preparing technology is simple and easy to carry out, the granularity of the prepared microsphere is even, the size is controllable, the storing stability is good, the leakage rate of the alcohol dehydrogenase in the using process is low, and the anti-acid-base environment capacity is strong.

The present invention provides methods for producing (S)-1,1,1-trifluoro-2-propanol, which include the step of reacting an enzyme of any one of alcohol dehydrogenase CpSADH, alcohol dehydrogenase ReSADH, carbonyl reductase ScoPAR, (2S,3S)-butanediol dehydrogenase ZraSBDH, carbonyl reductase ScGCY1, tropinone reductase HnTR1, tropinone reductase DsTR1, or alcohol dehydrogenase BstADHT, a microorganism or a transformant strain that functionally expresses the enzyme, or a processed material thereof, with 1,1,1-trifluoroacetone. The present invention also provides methods for producing (R)-1,1,1-trifluoro-2-propanol, which include the step of reacting alcohol dehydrogenase PfODH, a microorganism or a transformant strain that functionally expresses the enzyme, or a processed material thereof, with 1,1,1-trifluoroacetone.

The invention discloses a method for synthesizing a chiral bisaryl alcohol compound. The method provided by the present invention specifically uses a mutant of a carbonyl reductase SSCR derived from Sporobolomyces Salmonicolor and a mutant of alcohol dehydrogenase TbSADH derived from Thermoanaerobacter brockii to catalyze asymmetric reduction of (4-chlorophenyl)pyridine-2-ketone to form (S)-(4-chlorophenyl) pyridine-2-methanol. The experiment proves that the method provided by the invention has high yield and high ee value, can reduce the addition amount of coenzyme, and does not need to add enzyme such as glucose alcohol dehydrogenase for cofactor circulation, the reaction condition is mild, and the operation is simple.

A 3-hydroxypropionic acid(3HP) degradation way of halophilic bacteria is analyzed for the first time by a transcriptomics method, the inventor finds that the 3-hydroxypropionic acid can degrade genesdddA and dddC, and recombination halophilic bacteria TD delta dddA which cannot degrade 3HP is obtained. Based on this, an alcohol dehydrogenase gene AdhP and an aldehyde dehydrogenase gene AldDTD, which are from the halophilic bacteria endogenous and responsible for production and metabolism of the 3HP are further utilized, and a synthetizing biology means is taken for adjusting and controlling the expression level of the metabolic way. Through screening, the recombination halophilic bacteria high in yield of the 3HP is obtained, and the yield and the productivity are respectively 0.93g / g ofthe 1,3-propylene glycol(PDO) and 2.4g / L / h, which are the highest 3HP yield and productivity reported so far.

A novel gene having a function of improving acetic acid fermentation in practical level was cloned from a practical acetic acid bacterium belonging to the genus Gluconacetobacter by a method for obtaining a gene having a growth-promoting function in acetic acid-containing medium from the chromosomal DNA library of the acetic acid bacterium. It was made possible to significantly shorten the growth induction period and significantly improve the acetic acid fermentation rate of transformants obtained by introducing the gene into acetic acid bacteria, when such transformants are cultured in the presence of ethanol.

The invention provides an alcohol dehydrogenase LC3 and a gene and application thereof. An amino acid sequence of the alcohol dehydrogenase LC3 is shown in SEQ ID NO:2. An alcohol dehydrogenase gene lc 3 is extracted from lactobacillus curieae, a gene engineering strain is established, the heterologous expression of the alcohol dehydrogenase gene lc 3 is realized, and the enzymatic characteristic of the produced alcohol dehydrogenase LC3 is excellent. A co-expression gene engineering strain containing a glucose dehydrogenase gdh gene and the alcohol dehydrogenase gene lc3 is also successfully established, so as to realize the co-expression of glucose dehydrogenase and alcohol dehydrogenase. The alcohol dehydrogenase LC3 has the characteristics that the alcohol dehydrogenase LC3 and the gene engineering strain are applied to the production of chiral alcohol(R)-4-chloride-3hydroxy ethyl butyrate and other chiral alcohol products by a biological enzyme synthesizing method; the substrate concentration is high, the reaction rate is high, and the convenience is realized.

Chiral secondary alcohols may be produced enzymatically in high space-time yields while minimizing enzyme use, by reducing a keto compound in an aqueous reaction medium containing water, reducing agent, alcohol dehydrogenase and coenzyme, extracting the secondary alcohol formed by means of a further phase containing a water-immiscible organic solvent, and removing the phase used for extraction and reusing the aqueous reaction medium in step a).

The invention belongs to the technical field of biological genetic engineering, and particularly relates to a zymomonas mobilis recombination strain producing isobutanol, a construction method and application thereof. Zymomonas mobilis ZM4 serves as a model strain, firstly, the strain is transformed by means of genetic engineering, a metabolic intermediate in the synthesis pathway of valine of zymomonas mobilis is utilized for introducing a KDCA gene, 2-ketoisovalerate in the valine pathway is converted into a precursor isobutyraldehyde of isobutanol, and then isobutanol is generated under thehelp of alcohol dehydrogenase; secondly, the purpose of increasing the yield is achieved by overexpressing genes in the L-valine synthesis pathway and screening acetolactate synthase in the metabolicpathway;finally, by adjusting the expression quantity of different genes in the metabolic pathway, the purpose of high yield is finally achieved.

The invention relates to the field of molecular biology, and discloses a specific oligonucleotide probe for detecting an SNP (single nucleotide polymorphism) site of ALDH2 (alcohol dehydrogenase 2)gene, wherein the probe is capable of hybridizing with the 504th different genetype of the ALDH2 gene. The invention further discloses a specific primer pair for amplifying the ALDH2 gene when detecting the SNP site, and the primer pair can be used for specifically amplifying a target area containing a detection target site in the ALDH2 gene. The specific oligonucleotide probe and the specific primer pair provided by the invention can be used for detecting the ALDH2 genetype and providing information for people to learn individual alcohol metabolism detoxification ability and correctly use nitroglycerin.

The invention discloses an efficient preparation method of an intermediate of heterocyclic drugs. According to the efficient preparation method of the intermediate of the heterocyclic drugs, the intermediate of the heterocyclic drugs are produced by catalyzing a heterocyclic substrate by coupling an alcohol dehydrogenase mutant and glucose dehydrogenase; and the alcohol dehydrogenase mutant is prepared by mutating tyrosine at position 127 of a alcohol dehydrogenase female-parent with an amino acid sequence shown as SEQ ID No. 2 into tryptophan. The alcohol dehydrogenase mutant, namely mutant Y127W, utilized in the efficient preparation method of the intermediate of the heterocyclic drugs is capable of reducing product inhibition effects in a single aqueous system without addition of any co-solvent, thereby achieving a conversion rate higher than 99% within 12 hours. Being coupled with the glucose dehydrogenase (BmGDH), the mutant Y127W is capable of realizing gram-level preparation ata scale of 50 ml in a single aqueous system without addition of any exogenous coenzyme or organic co-solvent when substrate concentration is up to 600g x L<-1>; and the catalyst loading capacity is 3.3%. As for a final product, namely (S)-NBHP, the e.e. value is up to 99.4%; the space-time yield is about 1400g x L<-1> x d<-1>; and the product purity is 99.58%.