730 results about "Dehydrogenase" patented technology

The present invention relates to a recombinant yeast with high squalene production, a construction method and a use thereof, wherein the recombinant yeast is a saccharomyces cerevisiae that expresses or overexpresses HMGR gene, ADA gene, LEU2 gene, MVA pathway gene and squalene synthase gene ERG9. The present invention integrates acetaldehyde dehydrogenase gene ADA and HMG-CoA reductase gene tHMGR in multiple copies in saccharomyces cerevisiae, integrates the screened high-activity phosphoketolase gene PK and phosphotransacetylase gene PTA, weakens epoxysqualene synthesis, strengthens the mevalonate pathway, overexpresses yeast endogenous squalene synthase gene, etc. to obtain a recombinant yeast with high squalene production, so that the production in the shake flask stage of the production process exceeds 5g / L, and the production in the fed batch fermentation stage exceeds 50g / L, breaking through the current bottleneck of microbial synthesis of squalene production, and is suitable for practical promotion and application.

The invention relates to Pediococcus pentosaceus PP90 for preventing and treating alcoholic nerve injury and application of the Pediococcus pentosaceus PP90, the Pediococcus pentosaceus PP90 is preserved in China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC NO.32034. The Pediococcus pentosaceus PP90 can be used for preventing and treating alcoholic nerve injury. The fermentation product of the pediococcus pentosaceus PP90 can degrade the alcoholic strength of alcohol, can generate ethanol dehydrogenase and acetaldehyde dehydrogenase, can improve the activity of nerve cells, can regulate the expression of cytokines, can inhibit the activation of astrocytes, can regulate the expression of brain-derived neurotrophic factors, and can inhibit the growth of the brain-derived neurotrophic factors. The compound can improve the disordered neurotransmitter level, and can be used for preparing products for preventing or improving alcoholic nerve injury.

The invention relates to an enzyme reagent, glucose electrochemical test paper and a preparation method thereof, and belongs to the technical field of electrochemical detection. The glucose electrochemical test paper comprises an electrode substrate and an enzyme reagent arranged on the electrode substrate, the enzyme reagent is prepared from the following components in parts by mass: 5 to 7 parts of buffer solution, 0.01 to 0.2 part of hexamine ruthenium trichloride, 0.01 to 0.1 part of quinone compound, 0.1 to 1.2 parts of metal organic framework immobilized glucose dehydrogenase and 0.2 to 0.8 part of film-forming agent. According to the glucose electrochemical test paper, the detection accuracy of glucose can be improved, and the problems of large interference, small linear range, insensitive detection and the like in the prior art are improved or solved.

The invention discloses an alcohol dehydrogenase mutant and application of the alcohol dehydrogenase mutant in asymmetric synthesis of (S)-6-hydroxy-8-chloro ethyl caprylate with optical purity. The alcohol dehydrogenase or the mutant thereof provided by the invention has extremely high conversion rate and can effectively improve the optical purity of the product. Moreover, based on the mutant, low-cost, efficient and rapid production of single-configuration ethyl 6-hydroxy-8-chloro caprylate can be realized, a multi-configuration mixture is not generated, and the problems of tedious steps, pollution and the like caused by a conventional chemical synthesis method are also avoided.

The invention discloses a formate dehydrogenase mutant and application of the formate dehydrogenase mutant in catalysis of carbon dioxide reduction, the 223 arginine (Arg) of an amino acid sequence of formate dehydrogenase PsFDH48 derived from Paracoccus sp. MKU1 is mutated into proline (Pro) or the 242 proline (Pro) is mutated into phenylalanine (Phe) through a site-specific mutagenesis technology, and the obtained mutants R223P and P242F have higher catalytic activity and can be applied to catalysis of carbon dioxide reduction. The catalyst can catalyze CO2 to generate formic acid more effectively in vitro, and has potential application value in the aspects of efficient activation of CO2 and further conversion of CO2 into other mono-carbon compounds through cascade reaction.

The invention discloses an inositol dehydrogenase mutant and a preparation method of D-chiral inositol, and belongs to the technical field of genetic engineering. The inositol dehydrogenase mutant is obtained by mutating an amino acid sequence as shown in SEQ ID NO.2 through the following mutations: the 200th amino acid is mutated into C from V, the 234th amino acid is mutated into Q from V, and the 256th amino acid is mutated into E from R. According to the invention, the amino acid sequence of wild type inositol dehydrogenase is mutated to obtain the inositol dehydrogenase mutant which is a high-activity mutant capable of catalyzing conversion of myo-inositol into 2-keto-myo-inositol; therefore, when the inositol dehydrogenase mutant and keto isomerase act together to prepare D-chiral inositol by taking myo-inositol as a substrate, the yield of the D-chiral inositol is effectively improved.

The invention relates to the field of biochemical engineering, in particular to an alcohol dehydrogenase mutant and application thereof. The mutant is obtained by mutating alcohol dehydrogenase with an amino acid sequence encoding SEQ ID NO: 4 at one or more sites selected from 62nd, 108th, 130th, 132, 134, 182nd, 206th, 287th, 303th, 313th and 314, and the mutation is that the amino acid at the site is mutated into one of A, G, V, L, I, P, F, Y, W, S, T, C, M, N, Q, D, E, K, R and H. According to the alcohol dehydrogenase mutant and the application of the alcohol dehydrogenase mutant, the feeding amount of substrates is increased; the high conversion rate (gt; 99%) of the substrate is realized; according to the present invention, the new YqjM has the good product ee value (gt; 98%), and it is determined that the YqjM derived from the bacillus subtilis old yellow enzyme (belonging to ketene reductase) has the optimal catalytic activity, such that the good industrial application performance is provided.

The invention discloses a biological catalytic synthesis method of norambrox ether, which comprises the following steps: step 1, starting from trans-nerolidol, carrying out carroll rearrangement reaction to synthesize farnesyl acetone; 2, in the presence of a cofactor, converting farnesyl acetone into high farnesol acetate under catalysis of Bayer-Villiger monooxygenase; step 3, catalyzing the deacetylation of the high-farnesol acetate by using esterase to obtain EE-high-farnesol; and step 4, carrying out enzymatic conversion on farnesol by using the squalene hopaene cyclase mutant to obtain norambrox ether. The transformation process is safe and non-toxic, Bayer-Villiger monooxygenase which is new with a key enzyme, natural co-expressed esterase, alcohol dehydrogenase and squalene and hopaene cyclase mutants matched with the system are used for synergistic transformation, so that not only is the coenzyme for oxidation reaction promoted to be recycled, but also the conversion efficiency is improved; conversion from farnesyl acetone to norambrox ether can be efficiently and specifically catalyzed, and a final product can be directly obtained from a reaction system without separation and purification of an intermediate.

The invention discloses a medicinal and edible anti-alcoholism composition and a preparation method thereof, and relates to the crossing field of biotechnology and food and medicine, the composition comprises the following components: dried orange peel powder, vitamin C, glutathione, phellinus igniarius extract, polygonatum sibiricum extract, oriental wormwood extract, isatis root extract, schisandra chinensis extract, ginseng powder, lucid ganoderma extract and licorice extract. Silymarin, puerarin, white tomato powder, watermelon powder, fucoidin, tomato powder, mung bean powder, celery powder, turmeric powder, hawthorn fruit powder, honey powder, corn peptide, soybean peptide powder, xanthan gum and stevioside are added. The composition provided by the invention has the effect of increasing the content of ethanol dehydrogenase and acetaldehyde dehydrogenase in blood after drinking, and has the capability of promoting alcohol metabolism.

The invention belongs to the technical field of biology, and particularly relates to a method for producing xylitol through two-step enzyme catalysis of hemicellulose hydrolysate, and the method comprises the following steps: taking hemicellulose hydrolysate as a raw material, adding arabinose dehydrogenase, NADH oxidase and coenzyme NADH, and carrying out first-step fermentation until no L-arabinose residue exists in fermentation liquor to obtain fermentation reaction liquor I; and adding xylose reductase and glucose dehydrogenase into the fermentation reaction liquid I, and carrying out second-step fermentation to obtain a fermentation product containing xylitol. According to the two-step enzyme catalysis method provided by the invention, xylitol is prepared, and high-purity xylose does not need to be used as a raw material; according to the present invention, the cheap hemicellulose hydrolysate can be directly adopted as the raw material, the xylose conversion rate and the xylitol yield can exceed 99%, the impurity arabitol content in the product is low, the other impurity saccharic acid is easy to separate, and the problems of complex process, large separation difficulty and high cost in the xylitol refining process are successfully solved.

The invention discloses a 3-ketosteroid-delta1-dehydrogenase mutant and application thereof in synthesis of prednisone acetate, the mutant with high catalytic activity on cortisone acetate is obtained through screening, and a methanol-water phase (Tris-HCl buffer solution) system is adopted to increase the solubility of a substrate so as to improve the biological catalytic efficiency. In a single water phase system (Tris-HCl buffer solution) without methanol, the conversion efficiency is only 64.8% for 60g / L of cortisone acetate, and under the condition that other conditions are the same, the conversion rate in the Tris-HCl buffer solution system with methanol is increased to 92.1%, and the conversion rate is increased by 1.42 times.

The invention discloses alcohol dehydrogenase and a method for preparing (R)-6-hydroxy-8-chloro ethyl caprylate through catalysis of the alcohol dehydrogenase, and the alcohol dehydrogenase is selected from one or more of mutants of ADH1, ADH2, ADH3, ADH4, ADH5, ADH6, ADH7, ADH8, ADH9, ADH10 and ADH10; the amino acid sequences of the ADH1-ADH10 are as shown in SEQ ID NO. 1 to SEQ ID NO. 10. According to the invention, ethyl 6-carbonyl-8-chloro caprylate can be catalyzed to generate ethyl (R)-6-hydroxy-8-chloro caprylate or ethyl (S)-6-hydroxy-8-chloro caprylate, the reaction conversion rate of the mutant ADH10-A184L / N87F / A61L of ADH10 reaches 99%, and the e.e. Value of the R type is 99.2%.

The invention provides a formate dehydrogenase mutant with improved electron transfer rate and application thereof, the formate dehydrogenase mutant takes an amino acid sequence as shown in SEQ ID NO.1 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 97%, 98% or 99% identity with the SEQ ID NO.1 as a parent, and has site-saturated mutation corresponding to tyrosine at the 262nd site of the SEQ ID NO.1. The TsFDH mutant with improved electron transfer rate and stable catalytic activity is finally obtained by performing site-directed saturated mutation on formate dehydrogenase from thiobacillus.

The invention provides application of a lactobacillus mucus fermentation strain A21196 in preparation of a product for promoting ethanol metabolism, and belongs to the technical field of microbial preparations. The lactobacillus mucus fermentation strain A21196 has the high-yield ethanol dehydrogenase and acetaldehyde dehydrogenase, and has the effects of reducing the content of ethanol and acetaldehyde in the blood of mice suffering from acute alcoholism and shortening the central nervous inhibition time in vivo. Therefore, the invention provides the application of the fermented lactobacillus mucus strain A21196 in preparation of a product for preventing and / or treating the alcoholic liver injury or the alcoholic liver disease.

The invention discloses a Lactobacillus paracasei grx06 and an application thereof in the aspects of dispelling the effects of alcohol and protecting the liver, the strain number of the Lactobacillus paracasei is grx06, the Lactobacillus paracasei is preserved in the China General Microbiological Culture Collection Center on February 18, 2025, and the preservation number is CGMCC No: 33560. The lactobacillus paracasei obtained by screening in the invention has the capability of producing ethanol dehydrogenase and acetaldehyde dehydrogenase at high yield, and is high in ethanol tolerance. In addition, the traditional Chinese medicine composition has the effects of promoting ethanol metabolism in rats and recovering liver injury. Wide application prospects are realized in the field of anti-alcohol and liver-protecting products.

The invention provides a construction method and application of recombinant yarrowia lipolytica with high yield of tyrosol, and belongs to the field of bioengineering. The recombinant yarrowia lipolytica with high yield of tyrosol is named as a yarrowia lipolytica XJ-TL strain, and the recombinant yarrowia lipolytica with high yield of tyrosol is named as a yarrowia lipolytica XJ-TL strain. The recombinant yarrowia lipolytica disclosed by the invention expresses an ethanol dehydrogenase encoding gene, and simultaneously expresses any one or a combination of more of a DAHP synthase mutant encoding gene, a chorismate mutase mutant encoding gene, a glucose 6-phosphate dehydrogenase encoding gene, a transketolase encoding gene and a transaldolase encoding gene; meanwhile, a natural promoter of a pyruvate kinase coding gene is truncated. After the recombinant yarrowia lipolytica XJ-TL strain is subjected to fed-batch fermentation, tyrosol can be synthesized at a relatively high level. The construction method of the recombinant yarrowia lipolytica is easy to operate, and efficient synthesis of tyrosol can be achieved.

The invention discloses a recombinant pichia pastoris strain with high yield of retinaldehyde as well as a construction method and application of the recombinant pichia pastoris strain. The recombinant pichia pastoris strain is obtained by expressing dehydrogenase Zwf1, phosphate dehydrogenase Gnd2, NADH kinase Pos5 and two copied beta-carotene-15, 15 '-monooxygenase BCMO in host bacteria. Wherein the host bacterium is a pichia pastoris gene modified strain PP-B (Propene Polymer-B). The retinaldehyde production performance of the recombinant strain is verified based on cofactor engineering and the copy number of key enzymes, and the retinaldehyde production capacity of the pichia pastoris is further improved. The recombinant pichia pastoris disclosed by the invention is simple in construction method, can better promote the synthesis of retinol, and is beneficial to industrial production as the engineering strain can efficiently synthesize retinol by using methanol as a unique carbon source through amplification fermentation in a 5L fermentation tank.

The invention relates to pediococcus acidilactici PA27 capable of degrading alcohol and improving alcoholic nerve injury and application of the pediococcus acidilactici PA27, the pediococcus acidilactici PA27 is preserved in China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC NO.31922. The pediococcus acidilactici PA27 can degrade alcohol and improve alcoholic nerve injury. The pediococcus acidilactici PA27 separated and screened by the invention can grow and propagate in an alcohol-containing culture medium, can degrade alcohol and can generate ethanol dehydrogenase and acetaldehyde dehydrogenase; the polypeptide can be used for constructing a blood-brain barrier and reducing the damage of alcohol to central nervous cells; the traditional Chinese medicine composition can reduce the expression of inflammatory cytokines TNF-alpha, inhibit the activation of astrocytes, promote the expression of brain-derived neurotrophic factors BDNF, regulate the release of alcohol-induced astrocyte neurotransmitters and improve alcoholic nerve injury.

The invention discloses an enzyme catalytic synthesis method of a luliconazole chiral intermediate, which comprises the following steps: adding a recombinant escherichia coli wet cell for expressing alcohol dehydrogenase, a recombinant escherichia coli wet cell for expressing formate dehydrogenase, a buffer solution, a substrate, a cosolvent, ammonium formate and a coenzyme into a reaction container, reacting at 25-35 DEG C for 2-24 hours, extracting, separating, and carrying out rotary evaporation, thereby obtaining the luliconazole chiral intermediate. The luliconazole chiral intermediate is obtained; the substrate is 2, 2 ', 4'-trichloroacetophenone; the alcohol dehydrogenase is a mutant of AKR3. The method is mild in reaction condition, environmentally friendly, high in regioselectivity and stereoselectivity, high in conversion rate, high in chiral purity, small in enzyme dosage, low in preparation cost and suitable for industrial production.

A recombinant yeast cell that functionally expresses: —a nucleic acid sequence encoding a protein having glycerol dehydrogenase activity; —a nucleic acid sequence encoding a protein having dihydroxyacetone kinase activity; and —a nucleic acid sequence encoding a protein having glycerol transporter activity, wherein the expression of the nucleic acid sequence encoding the protein having glycerol transporter activity is under control of a promoter (the “GT promoter”), which GT promoter has an anaerobic / aerobic expression ratio for the glycerol transporter of 2 or more, and a process for the production of ethanol using such recombinant yeast cell.

This invention relates to the field of chicken gizzard lining processing technology, and particularly to an active polypeptide of chicken gizzard lining, its preparation method, and functional products. The preparation method includes the following steps: mixing chicken gizzard lining powder with water and subjecting it to alkaline heat treatment to form a slurry; sequentially adding Bacillus licheniformis alkaline protease, trypsin, and pepsin to the slurry for three separate hydrolysis processes to obtain a hydrolysate; after enzyme inactivation treatment of the hydrolysate, adding Lactobacillus plantarum for fermentation to obtain a fermentation broth; sterilizing the fermentation broth and then filtering it to obtain a fermentation supernatant; sequentially subjecting the fermentation supernatant to membrane filtration, sterilization, and spray drying to obtain the active polypeptide of chicken gizzard lining. The active polypeptide of chicken gizzard lining prepared by the method of this invention exhibits alcohol dehydrogenase activating activity, AGEs inhibitory activity, albumin denaturation inhibitory activity, lipoxygenase inhibitory activity, pancreatic lipase inhibitory activity, and a beneficial bacteria proliferation promoting effect.

The invention relates to the field of medicines, and particularly discloses application of mogroside V in preparation of a medicine for dispelling the effects of alcohol and protecting the liver, experiments prove that the anti-alcohol effect of the mogroside V is obviously superior to that of a mogroside crude extract, and in one embodiment, the mogroside V can increase protein expression of ethanol dehydrogenase and acetaldehyde dehydrogenase in the liver of a mouse at the same time. In one embodiment, the mogroside V can prolong the drunkenness tolerance time of a mouse and shorten the drunkenness time of the mouse. In another embodiment, the mogroside V can relieve liver inflammation caused by alcohol. In a further embodiment, the mogroside V can greatly relieve liver fat deposition caused by alcohol. Therefore, the mogroside V can be developed into a brand new product for dispelling the effects of alcohol and protecting the liver, and has a huge application prospect.

The invention discloses ethanol dehydrogenase, recombinant genetically engineered bacteria and application of the ethanol dehydrogenase in asymmetric reduction of prochiral carbonyl compounds, the ethanol dehydrogenase has good biological performance, heterologous expression in Escherichia coli engineering bacteria is achieved, intracellular coenzyme can be directly utilized when whole cells are adopted as a catalyst, and the catalyst has a good application prospect. Zero addition of the coenzyme is realized, the enzyme has the advantages of high activity, high stereoselectivity, wider substrate spectrum and the like, prochiral ketone compounds can be greenly and efficiently catalyzed to be converted into corresponding chiral alcohols, and the problems of low conversion rate, low yield, low e.e. Value, complex reaction process, more byproducts and the like in a traditional chemical method are solved. The ethanol dehydrogenase has good tolerance to isopropanol, and the isopropanol can be used as a hydrogen source to realize the circulation of coenzyme. And the byproduct acetone generated by the enzyme catalytic reaction and the isopropanol remaining in the reaction can be separated and recovered by a technical means, so that the production cost is greatly reduced.

The invention belongs to the technical field of enzyme catalysis, and particularly relates to a dehydrogenase mutant, a gene, an engineering bacterium and application, the dehydrogenase mutant is obtained through mutation of original dehydrogenase, the sequence of the original dehydrogenase is SEQ ID NO.1, and the mutation mode is that the 344 site is mutated from A to W, K, F, S or N, or the 130 site is mutated from H to T, W or F; the dehydrogenase mutation diagram provided by the invention has higher conversion rate, stronger specificity and faster reaction speed.

The invention belongs to the technical field of medicines, and particularly relates to a plant lactobacillus for relieving acute alcoholic liver injury as well as a metagen and application of the plant lactobacillus. The strain of the plant lactobacillus L5-13 is preserved in the China Center for Type Culture Collection on August 18, 2025, and the preservation number of the strain is CCTCC NO: M 20251835. The strain and the metagen thereof can significantly improve pathological symptoms of mice with acute alcoholic liver injury, including: reducing the levels of ALT, AST, TC, TG and LDL-C in serum of the mice with acute alcoholic liver injury, and increasing the level of HDL-C; the abnormal blood lipid level of the mouse caused by acute alcohol is regulated; the activity of antioxidant enzymes in the liver is improved, and the lipid peroxidation level is reduced; the ethanol dehydrogenase activity of the liver is improved, and the ethanol metabolism capability is enhanced. Therefore, the strain and the metagen thereof have wide application prospects in prevention or treatment of acute alcoholic liver injury.

The invention discloses a phosphite dehydrogenase mutant, a biological material, a preparation method and application, and compared with wild phosphite dehydrogenase, the phosphite dehydrogenase mutant has one or more amino acid residue mutations in the 29th site, the 31st site, the 59th site, the 68th site, the 185th site and the 275th site. According to the mutant, the regeneration efficiency of reduced coenzyme NADH is effectively improved, and the catalytic efficiency of the mutant N29T / Q185C / K275E is 1.7 times that of wild type phosphite dehydrogenase; the mutant is mild in reaction condition and wide in range, the reaction temperature is 15-40 DEG C, the reaction pH is 4-10, the stability is improved compared with that of wild phosphite dehydrogenase, and the mutant has high application value.

The invention belongs to the technical field of biochemical engineering, and particularly relates to a marine metagenome sourced meso-diaminopimelate dehydrogenase mutant and application thereof in D-tryptophan synthesis. According to the invention, a fast evolution method based on Rosetta guidance is adopted to modify meso-diaminopimelate dehydrogenase from a marine metagenome, and a mutant with high catalytic activity on 3-indolepyruvic acid is obtained through screening. The meso-diaminopimelate dehydrogenase mutant disclosed by the invention can be applied to direct catalysis of reductive amination of 3-indolepyruvic acid to generate chiral D-tryptophan, and can also be coupled with L-amino acid deaminase and coenzyme regeneration enzyme to realize configuration inversion from L-tryptophan to D-tryptophan, the substrate concentration reaches 0.1 M, and the product conversion rate is gt; the ee value is gt; therefore, the method has a relatively high practical application value.

The invention discloses an aldose reductase mutant and application thereof in production of D-tagatose, and belongs to the technical field of genetic engineering. The aldose reductase mutant disclosed by the invention has relatively high catalytic activity, and can be coupled with galactitol 2-dehydrogenase to realize cyclic regeneration of NADH (nicotinamide adenine dinucleotide), so that the preparation cost of D-tagatose is greatly reduced. When the aldose reductase mutant is used for preparing D-tagatose, 98.2 g / L of D-tagatose can be obtained through conversion within 5 h, the yield reaches up to 98.2%, and the conversion effect is better than that of wild type and other mutant enzymes, so that the aldose reductase mutant has good industrial application prospects.

The invention discloses an Escherichia coli endogenous alcohol dehydrogenase EcYjgB mutant and application thereof. The mutant is obtained by performing any one of the following mutations (a) to (c) on Escherichia coli endogenous alcohol dehydrogenase EcYjgB as shown in SEQ ID NO.1: (a) mutating serine at the 200th site into arginine; (b) the serine at the 199th site is mutated into aspartic acid, and the serine at the 200th site is mutated into arginine; and (c) the serine at the 199th site is mutated into aspartic acid, the serine at the 200th site is mutated into arginine, and the asparagine at the 240th site is mutated into tryptophan. The mutant is used for efficiently catalyzing reduction of HMF or furfural to respectively synthesize BHMF and furfuryl alcohol, so that the purposes of improving the production efficiency, reducing the production cost and improving the environmental effect are achieved.

The invention discloses a separation and preparation method of scyllo-inositol, and relates to the technical field of scyllo-inositol production, myo-inositol solution is added into whole-cell wet thalli containing inositol dehydrogenase and scyllo-inositol dehydrogenase, conversion is carried out in PBS buffer solution to obtain conversion liquid, the conversion liquid is filtered and decolorized by a ceramic membrane and an ultrafiltration membrane, and the scyllo-inositol is obtained. Then concentrating and centrifuging to obtain a mixed crude product of scyllo-inositol and inositol, pulping and washing with purified water, and drying a wet product to obtain scyllo-inositol. In the separation and preparation process, boric acid, strong acid, strong alkali and organic solvent are not used, precipitation of composite salt is not needed, the process steps are reduced, the environment is not polluted, the operation is safe, the recovery cost is low, materials are recycled, a complex recovery system is not needed, the cost can be better reduced, and the method is environment-friendly.