25 results about "Alcohol dehydrogenase" patented technology

The application discloses a method for synthesizing high-concentration furfuryl alcohol by using a double-enzyme coupling method, and the method comprises the following steps: mixing alcohol dehydrogenase freeze-dried bacterial powder and glucose dehydrogenase freeze-dried bacterial powder to serve as a catalyst, using furfural as a substrate, using glucose as a co-substrate, using NADP + or NAD + as a coenzyme, using a buffer solution with a pH value of 4-9 as a reaction medium to form a reaction system, and reacting completely under the condition of 20-50 DEG C and 400 rpm, then separating and purifying the reaction liquid to obtain furfuryl alcohol. The biological catalytic system is suitable for furfural with a concentration higher than 500 mM, and 100% conversion of 500 mM can be realized within 10 hours. The substrate continuous flow process is further used to eliminate the inhibition of aldehyde substrates on the biological catalyst, the concentration of the product can reach 1500 mM, and the highest space-time yield can reach 235 g / (L.d).

The present application provides a kind of kudzu vine cranberry composition for protecting liver from alcohol, which contains the following ingredients by weight: 30-50 parts of extract of Hovenia dulcis Thunb, 30-50 parts of mixed extract, 20-40 parts of extract of Pueraria lobata, 10-20 parts of extract of Poria cocos, 10-20 parts of extract of Glycyrrhiza uralensis, 0.05-0.1 parts of vitamin C; the raw materials of the mixed extract are mixed by cranberry and Fructus Lycii in a mass ratio of (1-2):(1-2). The present application further provides formulations and food containing the composition. The present application further protects the use of the composition or formulation in preparing health food or medicine with auxiliary protective effect on chemical liver injury. The composition provided by the present application can improve the activity of alcohol dehydrogenase / acetaldehyde dehydrogenase, reduce the content of MDA in tissues and increase the content of GSH, and reduce the content of triglyceride in liver cells, thereby comprehensively exerting the effect of protecting liver from alcohol, with high safety, small side effects, and wide sources of raw materials, and having important market value and development prospect.

The application discloses an alcohol dehydrogenase mutant and application thereof in enzymatic synthesis of a loratadine intermediate. A plurality of alcohol dehydrogenase mutants are developed, and a loratadine chiral intermediate is obtained through an enzyme catalytic synthesis method based on the mutants, wherein the reaction condition is mild, the environment is friendly, the reaction has high regional selectivity and stereoselectivity, the reaction conversion rate is high, the chiral purity of the product is high, the enzyme consumption is low, the preparation cost is low, the method is suitable for industrial production, the method avoids the problems of difficult chiral resolution and heavy metal residue in a product in a conventional method, and the method makes up for the deficiencies of a traditional chemical method.

The embodiment of the present application discloses a Clostridium ethanoligenes mutant with alcohol-aldehyde dehydrogenase knocked out and application thereof in reducing reverse absorption of ethanol product in one-carbon gas fermentation process, and belongs to the technical field of biotechnology. In the process of producing ethanol and microbial protein by Clostridium ethanoligenes using one-carbon gas fermentation, the reverse absorption phenomenon of product ethanol, the alcohol-aldehyde dehydrogenase (Adh, Alcohol dehydrogenase) on the chromosome of Clostridium ethanoligenes is knocked out by gene editing means, so as to reduce or completely block the reverse absorption pathway of ethanol, reduce the loss of ethanol in the fermentation process and in the wort storage tank, and help to improve the ethanol yield, reduce the waste of carbon source and improve the enterprise income.

The application discloses an alcohol dehydrogenase mutant and application thereof in preparation of cis-4-propylcyclohexanol. The mutant is obtained by substituting at least one amino acid in three positions of an amino acid sequence shown in SEQ ID No. 1, i.e. the 43th, 271th and 282th positions. The 43th phenylalanine (F) is substituted into valine (V) or serine (S), the 271th isoleucine (I) is substituted into valine (V) or phenylalanine (F), and the 282th phenylalanine (F) is substituted into valine (V). The wild-type alcohol dehydrogenase gene is subjected to directional evolution modification, and the alcohol dehydrogenase mutant has higher catalytic activity and higher stereoselectivity than the wild-type alcohol dehydrogenase, so that the cis-4-propylcyclohexanol is efficiently and highly stereoselectively generated.

The present invention belongs to the technical field of genetic engineering. Provided are an enzyme formulation for the catalytic synthesis of scyllo-inositol, and a method for preparing scyllo-inositol. The enzyme formulation has the enzymatic activity of catalyzing the reaction of myo-inositol to form scyllo-inositol, and comprises inositol dehydrogenase and scyllo-inositol dehydrogenase. After mutation, inositol dehydrogenase has a higher enzymatic activity, and scyllo-inositol dehydrogenase has a broader applicable pH range. Further provided is a method for preparing scyllo-inositol, which method comprises: (1) preparing a crude enzyme solution of inositol dehydrogenase and scyllo-inositol dehydrogenase, preparing a myo-inositol solution, and adding the myo-inositol solution to the crude enzyme solution; (2) adjusting a pH value to 7-10, controlling a temperature to 40-50°C, and performing a conversion reaction to obtain conversion solution A; and (3) subjecting conversion solution A to first cooling crystallization and filtration, and collecting a filtrate and a filter cake; adding water and a myo-inositol solid to the filtrate, and repeating step (2) and step (3) until no crystals precipitate; and subjecting the filter cake collected each time to first ethanol washing, second cooling crystallization, filtration, second ethanol washing and drying in sequence, to obtain scyllo-inositol. The method for preparing scyllo-inositol has good conversion performance, shortens the production cycle of scyllo-inositol, and reduces waste liquid generation and environmental pollution.

This invention discloses an enzymatically catalytic synthesis method for a chiral intermediate of dzoxadamine. Using methyl acetoacetate as a substrate, buffer, substrate, alcohol dehydrogenase, formate dehydrogenase, ammonium formate, and coenzyme are added to a reaction vessel. The reaction is carried out at 20–40°C for 2–24 hours. After extraction, separation, and rotary evaporation, the chiral intermediate (R)-3-hydroxybutyrate methyl ester is obtained. The alcohol dehydrogenase is a mutant of ADH1 derived from Rollstonella, and the amino acid sequence of ADH1 is shown in SEQ ID No. 1. This invention offers mild reaction conditions, high stereoselectivity, and environmental friendliness. It also boasts high conversion rate, high chiral purity, low enzyme dosage, tolerance to high substrate concentrations, and low preparation cost, making it suitable for industrial production. Furthermore, it avoids chiral resolution and heavy metal residues in the product, overcoming the shortcomings of chemical methods.

This invention provides a fermenting *Lactobacillus mucinus* strain that can accelerate alcohol metabolism and improve hangover symptoms, along with its applications. This strain exhibits high safety, lacking resistance genes and pathogenic virulence genes, and demonstrates strong tolerance to bile salts, artificial gastric juice, and artificial intestinal juice. The bacterium possesses strong antioxidant capabilities, exhibiting potent scavenging abilities against ABTS, DPPH, and hydroxyl radicals. This fermenting *Lactobacillus mucinus* can increase the alcohol dehydrogenase content in a mouse model of alcoholic liver disease, demonstrating significant application value in antioxidation, alcohol detoxification, and hangover prevention. Furthermore, this bacterium can produce melatonin and convert NR to niacin.

The application discloses a genetically engineered bacterium for improving the yield of mevalonic acid as well as a construction method and application thereof, and belongs to the technical field of genetic engineering. In order to solve the technical problems of low carbon atom economy and limited theoretical yield of MVA biosynthesis technology, the application takes Escherichia coli as a starting strain, overexpresses a bifunctional acetaldehyde-ethanol dehydrogenase mutant AdhE A267T / E568K gene adhE mut , an acetyl-CoA acyltransferase / HMG-CoA reductase gene mvaE , an HMG-CoA synthetase gene mvaS , and introduces a tetracycline-induced CRISPRi system to inhibit the expression of an orotate-5'-phosphate decarboxylase gene pyrF , so that the carbon flux is led from the growth of the strain to the synthesis of mevalonic acid. The genetically engineered bacterium provided by the application can produce mevalonic acid by fermentation with ethanol as a carbon source, and the yield reaches 0.89 g / g. The application provides a new idea for improving the carbon atom economy of mevalonic acid biological manufacturing.

The application discloses a kind of for waste PET depolymerization system glycol directional efficient conversion of glycolic acid two enzyme level connection system and its application, belong to biological catalysis and resource recycling technical field.The system includes a kind of mutant alcohol dehydrogenase with high selectivity to glycol and a mutant aldehyde dehydrogenase ALDH-V7.Said mutant alcohol dehydrogenase is obtained in at least one combination mutation in 97 and 101 and / or 145 and 148 in the basis of ADH5 enzyme derived from acetic acid bacillus ( Gluconobacter oxydans ).The system can efficiently and directionally oxidize low-value glycol in waste PET chemical or biological depolymerization product to high-value-added product glycolic acid under mild conditions, effectively avoiding peroxidation and C-C bond rupture.The application provides an efficient and green biological catalysis solution for upgrading and recycling of waste PET, and realizes a circular economy path of "waste plastics-degradable plastic monomers".

The application belongs to the field of bioengineering and biocatalysis technology, and particularly relates to an alcohol dehydrogenase mutant and application thereof. The application discloses an alcohol dehydrogenase mutant CcPAR-M1 (V158L), and an amino acid sequence of the mutant is shown as SEQ ID No. 3. The mutant can be used for catalyzing asymmetric reduction reaction of a biaryl aldehyde substrate through dynamic kinetic resolution to generate R an axially chiral biaryl dimethanol compound. By using the biocatalytic system disclosed in the application, the highest yield of the target product can reach 96%, and the enantiomeric excess value can reach 99% ee at most. The method provides a green, efficient and excellent stereoselective biocatalytic strategy for construction of the axially chiral biaryl dimethanol compound.

This invention provides a two-phase aqueous extraction apparatus and a method for simultaneously extracting alcohol dehydrogenase and heparin from pig liver, belonging to the field of chemical manufacturing technology. The two-phase aqueous extraction apparatus includes two storage cylinders; each storage cylinder has an independently mounted rotating shaft; a driving wheel is fixedly mounted on each rotating shaft; a first driven wheel is mounted on the left side of the driving wheel; a first horizontal shaft is fixedly mounted at the center of the first driven wheel; the first horizontal shaft is rotatably mounted on the inner wall of the storage cylinder; a first stirring rod is evenly arranged on the outer wall of the first horizontal shaft; a second driven wheel is engaged on the right side of the driving wheel; a second horizontal shaft is fixedly mounted at the center of the second driven wheel. Using this apparatus to extract from pig liver is not only efficient, simple, and environmentally friendly, but also allows for the simultaneous extraction of highly active alcohol dehydrogenase and high-purity heparin. This invention provides new ideas for related fields such as the food and pharmaceutical industries.

The present application belongs to the field of bioengineering technology, and particularly relates to an alcohol dehydrogenase YugJ mutant and its application in fermentative production of tyrosol or hydroxytyrosol. The present application adopts a method of site-directed mutation to mutate the serine at the 41st position, the threonine at the 138th position and the valine at the 183rd position of alcohol dehydrogenase YugJ from Bacillus licheniformis into proline, serine and leucine respectively to obtain a mutant PSI (S41P / T138S / V183I). The mutant PSI improves the enzyme activity of alcohol dehydrogenase, effectively solves the problem of low alcohol performance of alcohol dehydrogenase at present, and is successfully applied to the fermentative production of tyrosol and / or hydroxytyrosol, thereby providing a new idea for improving the production level of alcohol compounds.

The application discloses a plant attenuated mutant and application thereof in production of exogenous proteins, and belongs to the technical field of biology. The attenuated mutant is that the 182th amino acid lysine of a proteinase of a zucchini yellow mosaic virus auxiliary component is changed into arginine. The attenuated mutant of the zucchini yellow mosaic virus can be used for efficiently producing alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH), and the plant used for production does not appear necrosis.

This invention discloses a genetically engineered bacterium and its application in enhancing lactic acid production in *Escherichia coli*, belonging to the field of bioengineering. The genetically engineered bacterium of this invention uses *E. coli* as the starting strain and is obtained through genetic engineering operations. In this genetically engineered bacterium, the phosphoacetyltransferase-acetylkinase locus pta-ack and the alcohol dehydrogenase gene adhE are knocked out or deleted, while exogenous lactate dehydrogenase and exogenous monocarboxylic acid transporter genes are introduced and overexpressed. The modified genetically engineered bacterium, while retaining the advantages of *E. coli* such as easy culture and rapid growth rate, significantly improves the yield and conversion rate of lactic acid. Compared to the starting strain, the lactic acid concentration increases to 6 g / L with pta-ack knockout alone, and to 8 g / L with double knockout of pta-ack and adhE. Furthermore, the lactic acid concentration increases to approximately 20 g / L and 23 g / L with overexpression of exogenous lactate dehydrogenase and exogenous monocarboxylic acid transporter, respectively.

The application belongs to the technical field of enzyme engineering and bioengineering, and particularly relates to a mutant alcohol dehydrogenase, a freeze-dried powder thereof and application of the mutant alcohol dehydrogenase in preparation of (2S, 3S)-2-hydroxy-4-phenylbutane derivatives. The mutant alcohol dehydrogenase of the application has a reduced enzyme dosage by half for achieving the same conversion rate and stereoselectivity in catalyzing synthesis of (2S, 3S)-2-hydroxy-4-phenylbutane derivatives, and shows higher catalytic efficiency. The enzyme amorphous powder obtained by freeze-drying is easy to store and transport for a long time, and can be directly weighed and fed in use, which is convenient to operate. The preparation method of the application uses high-concentration isopropyl alcohol (60% m / V) as a cosolvent and a coenzyme regeneration system, and does not need to additionally add glucose and glucose dehydrogenase, thereby simplifying a process flow, reducing a cost, and achieving a short reaction time (16-18 h), a high conversion rate (up to 99.8% or more), and excellent stereoselectivity (de value > 99.9%), which is very suitable for industrial production.

This paper discloses the use of modified Escherichia coli and its L-amino acid fermentation production. Specifically, it discloses the use of pyruvate formate lyase, alcohol dehydrogenase, branched-chain amino acid transaminase derived from Escherichia coli, branched-chain amino acid transaminase derived from Bacillus subtilis, thiamine phosphate synthase, and maleate dehydrogenase in the construction of genetically modified bacteria that produce L-amino acids. Experiments have shown that by knocking out the genes encoding pyruvate formate lyase, alcohol dehydrogenase, branched-chain amino acid transaminase derived from Escherichia coli, thiamine phosphate synthase, and maleate dehydrogenase in E. coli capable of producing L-amino acids, and introducing the gene encoding branched-chain amino acid transaminase derived from Bacillus subtilis, recombinant bacteria favorable for L-amino acid accumulation can be obtained. These recombinant bacteria can significantly increase L-amino acid production and reduce costs, which is of great significance in accelerating the industrialization process of L-amino acids.

This invention discloses an alcohol dehydrogenase mutant and its application in the asymmetric synthesis of optically pure (S)-6-hydroxy-8-chlorooctanoate ethyl ester. The alcohol dehydrogenase or its mutant in this invention exhibits extremely high conversion rates and effectively improves the optical purity of the product. Furthermore, based on this mutant, low-cost, efficient, and rapid production of a single configuration of 6-hydroxy-8-chlorooctanoate ethyl ester is achieved, avoiding the generation of multi-configuration mixtures and the problems of cumbersome steps and contamination associated with conventional chemical synthesis methods.

The present invention relates to a method for preparing a glycolic acid containing aqueous solution, wherein ethylene glycol, which is present in the form of an aqueous solution, is oxidized in vitro to glycolaldehyde by treatment with a first NAD-dependent oxidoreductase and NAD + + to glycolic acid, and the reduced NADH resulting from these oxidations is re-oxidized and thereby regenerated by a NADH oxidase or an alcohol dehydrogenase.​

The present application relates to the technical field of genetic engineering, in particular to an ethanol-responsive genetically engineered alcoholism-relieving probiotic, a construction method and application thereof. The probiotic provided by the present application introduces an "ethanol-responsive" regulation mechanism, so that the strain can specifically induce the efficient expression of human alcohol dehydrogenase 1B (hADH1B) and human acetaldehyde dehydrogenase 2 (hALDH2) when sensing the presence of ethanol in the body, thereby realizing the on-demand and efficient decomposition of ethanol and its toxic metabolite acetaldehyde. Compared with the continuous expression of traditional engineering bacteria, this intelligent regulation mode can better simulate the natural metabolic rhythm of the human body, solve the problem of bacterial metabolism, improve the alcoholism-relieving efficiency, reduce the metabolic pressure of the bacterial cells, and is expected to achieve better alcoholism-relieving effect and higher biological safety.

The application belongs to the field of green manufacturing, and discloses a preparation method of a pyridazine compound, comprising the following steps: (1) adding an enzyme catalyst, a bio-based furan and a co-substrate into a buffer solution to react, and removing cells after the reaction; (2) adding hydrogen peroxide or an oxidase and a titanium-silicon molecular sieve to react, and removing the titanium-silicon molecular sieve after the reaction; (3) slowly adding hydrazine under stirring at 0-60 DEG C to obtain the pyridazine compound after the reaction; the enzyme catalyst is at least one of the following: an alcohol dehydrogenase / glucose dehydrogenase double enzyme system, an alcohol dehydrogenase / formate dehydrogenase double enzyme system and a recombinant bacterium co-expressing one or both of the above two double enzyme systems. The application has the advantages of high selectivity, mild reaction conditions, environmental friendliness, fast reaction speed and no need for intermediate separation and purification.

This invention discloses an alcohol dehydrogenase derived from *Rhodotorula toruloides* and its application in the asymmetric reduction of 1-(2-chlorophenyl)-2-(1,2,3,4-tetrazol-2-yl)ethane-1-one to (R)-1-(2-chlorophenyl)-2-(1,2,3,4-tetrazol-2-yl)ethane-1-ol. The alcohol dehydrogenase can catalyze substrate conversion reactions at concentrations greater than 400 g / L, with a conversion rate >99.9% and a product ee value >99.5%. The reaction rate is fast, easily controlled, and the catalytic level is significantly improved compared to previous reports, demonstrating great potential for industrial application.

This invention discloses an NADPH-dependent alcohol dehydrogenase EcYjgB mutant and its applications. The mutant is based on the wild-type alcohol dehydrogenase EcYjgB with mutations occurring at one or more of the following sites: S46, E51, W52, F54, T92, I108, A114, P116, I118, N240, G262, V264, L265, and A286. The amino acid sequence of the wild-type alcohol dehydrogenase EcYjgB is shown in SEQ ID. 1. This mutant is used to catalyze the selective reduction of high concentrations of 5-hydroxymethylfurfural or furfural to synthesize 2,5-furandiethanol or furfuryl alcohol, respectively. Compared to the wild-type enzyme, this mutant exhibits significantly improved catalytic efficiency and significantly reduced substrate inhibition at high substrate concentrations. It is green and efficient, requires no organic reagents, shortens reaction time, and significantly reduces the difficulty and cost of subsequent separation and purification.

This invention discloses a *Stachys pubescens* alcohol dehydrogenase, its encoding gene, and its applications, relating to the field of microbial genetic engineering technology. The alcohol dehydrogenase is any one of (1), (2), and (3): (1) the SvADH1 protein, whose amino acid sequence is shown in SEQ ID NO.3; (2) a protein obtained by substituting, deleting, and / or adding one or more amino acid residues to the amino acid sequence of the SvADH1 protein, resulting in a protein with more than 70% identity to the SvADH1 protein and associated with the redox of cross-linked solanol A; (3) a fusion protein obtained by attaching a protein tag to the N-terminus and / or C-terminus of (1) or (2). This invention has found that overexpression of the encoding gene of this alcohol dehydrogenase in *Stachys pubescens* significantly increases the yield of cross-linked solanol A, thus providing a new direction for the development of microbially derived cross-linked solanol A.