746 results about "Reductase" patented technology

NADP-dependent oxidoreductase compositions, and electrodes, sensors and systems that include the same. Analyte sensors include an electrode having a sensing layer disposed thereon, the sensing layer comprising a polymer and an enzyme composition distributed therein. The enzyme composition includes nicotinamide adenine dinucleotide phosphate (NAD(P)+) or derivative thereof, an NAD(P)+-dependent dehydrogenase; an NAD(P)H oxidoreductase; and an electron transfer agent comprising a transition metal complex.

NADP-dependent oxidoreductase compositions, and electrodes, sensors and systems that include the same. Analyte sensors include an electrode having a sensing layer disposed thereon, the sensing layer comprising a polymer and an enzyme composition distributed therein. The enzyme composition includes nicotinamide adenine dinucleotide phosphate (NAD(P)+) or derivative thereof, an NAD(P)+-dependent dehydrogenase; an NAD(P)H oxidoreductase; and an electron transfer agent comprising a transition metal complex.

NADP-dependent oxidoreductase compositions, and electrodes, sensors and systems that include the same. Analyte sensors include an electrode having a sensing layer disposed thereon, the sensing layer comprising a polymer and an enzyme composition distributed therein. The enzyme composition includes nicotinamide adenine dinucleotide phosphate (NAD(P)+) or derivative thereof; an NAD(P)+-dependent dehydrogenase; an NAD(P)H oxidoreductase; and an electron transfer agent comprising a transition metal complex.

NADP-dependent oxidoreductase compositions, and electrodes, sensors and systems that include the same. Analyte sensors include an electrode having a sensing layer disposed thereon, the sensing layer comprising a polymer and an enzyme composition distributed therein. The enzyme composition includes nicotinamide adenine dinucleotide phosphate (NAD(P)+) or derivative thereof; an NAD(P)+-dependent dehydrogenase; an NAD(P)H oxidoreductase; and an electron transfer agent comprising a transition metal complex.

The invention discloses a biosynthesis system for high-yield 1, 6-hexamethylenediamine and application of the biosynthesis system, and belongs to the technical field of biology. According to the method, key genes which are excavated and optimized in a 1, 6-hexamethylenediamine biosynthetic pathway are co-expressed in a single cell module or a multi-cell module, and different enzyme elements expressed by the cell module are subjected to enzyme cascade reaction, so that 1, 6-hexamethylenediamine can be efficiently synthesized by taking caprolactam or 6-aminocaproic acid as a substrate; furthermore, after the carboxylic acid reductase MaCAR is mutated, the catalytic activity of the carboxylic acid reductase MaCAR can be remarkably improved, and the yield of the 1, 6-hexamethylenediamine is further improved; in addition, the biosynthesis system disclosed by the invention directly takes the cell module as a whole-cell catalyst, and an enzyme purification step is avoided, so that the biosynthesis system has the advantage of low cost.

NADP-dependent oxidoreductase compositions, and electrodes, sensors and systems that include the same. Analyte sensors include an electrode having a sensing layer disposed thereon, the sensing layer comprising a polymer and an enzyme composition distributed therein. The enzyme composition includes nicotinamide adenine dinucleotide phosphate (NAD(P)+) or derivative thereof, an NAD(P)+-dependent dehydrogenase; an NAD(P)H oxidoreductase; and an electron transfer agent comprising a transition metal complex.

NADP-dependent oxidoreductase compositions, and electrodes, sensors and systems that include the same. Analyte sensors include an electrode having a sensing layer disposed thereon, the sensing layer comprising a polymer and an enzyme composition distributed therein. The enzyme composition includes nicotinamide adenine dinucleotide phosphate (NAD(P)+) or derivative thereof; an NAD(P)+-dependent dehydrogenase; an NAD(P)H oxidoreductase; and an electron transfer agent comprising a transition metal complex.

The invention discloses a mesophilic carotenoid-producing recombinant yarrowia lipolytica strain as well as a construction method and application thereof, the recombinant strain exogenously expresses mesophilic original fatty acid desaturase (FAD3) and isocitrate dehydrogenase (IDH2) mutants from phaffia rhodozyma, and efficient synthesis under the condition of 30 DEG C is realized; secondly, a metabolic pathway of the recombinant strain is optimized based on a metabolic flow push-pull strategy, and isopentenyl pyrophosphate isomerase (IDI), farnesyl pyrophosphate synthase (ERG20), plant alkene dehydrogenase (CarB) and bifunctional plant alkene synthase (CarRP) are overexpressed, so that the flux of a precursor is increased; finally, gene multi-copy of astaxanthin synthase (crtS) and cytochrome P450 reductase (crtR) from phaffia rhodozyma is carried out, and the carotenoid production capacity of yarrowia lipolytica is further improved. The construction method of the mesophilic yarrowia lipolytica is simple, efficient synthesis of carotenoid can be better achieved at the higher temperature, the fermentation cost is greatly reduced, and the industrial production potential is achieved.

Disclosed are, inter alia, double stranded RNAi (dsRNAi) agents inhibiting expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), for example, human HMGCR, compositions including the same, and methods of treatment using the same.

The invention relates to the technical field of biochemical engineering, in particular to a method for preparing chiral alpha-trifluoromethyl alcohol by using imine reductase AtRedAm, trifluoromethyl ketone is reduced into corresponding chiral trifluoromethyl alcohol at an excellent conversion rate by using the imine reductase AtRedAm, and enantioselectivity is high. The reaction depends on an NADPH (Nicotinamide Adenine Dinucleotide Phosphate) coenzyme circulation system, 10% dimethyl sulfoxide is used as a reaction cosolvent, the reaction is carried out in a 100mM (pH = 7.5) phosphate buffer solution for 16 hours, the highest molar conversion rate reaches 99%, and the highest optical purity reaches 95%.

The invention relates to the field of biological engineering and technology, in particular to an enzyme for synthesizing glabridin, a biological synthesis method of glabridin and a bacterial strain. The invention discloses a series of glabridin synthesizing enzymes (phenylalanine ammonialyase, cinnamic acid-4-hydrogenase, tyrosine ammonialyase, p-coumaric acid coenzyme A ligase, chalcone synthase, chalcone reductase, chalcone isomerase, isoflavone synthase, 2-hydroxyisoflavone dehydratase, 4 '-oxymethyltransferase, 4'-hydroxymethyltransferase and 4 '-hydroxymethyltransferase) derived from glycyrrhiza glabra for the first time. ) can be used in the preparation of a compound (e.g., isoflavone 2 '-hydrogenase, isoflavone reductase, Vissisoketoreductase, isoprenyltransferase, pterocarpin reductase, oxidative cyclase, demethylase, cytochrome P450 reductase, and the like). The de novo synthesis of the glabridin in a microbial cell factory is realized for the first time, and the application prospect of the glabridin biological manufacturing industry is greatly promoted.

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 dihydrodaidzein reductase mutant and application of the dihydrodaidzein reductase mutant in synthesis of (3S, 4R)-tetrahydrodaidzein, and belongs to the technical field of biology. According to the mutant, wild-type dihydrodaidzein reductase DHDR is taken as a parent, single-point mutation or combined mutation is carried out, and a DHDR mutant is obtained; the parent site of the single point mutation is one of the 120th site, the 122nd site, the 170th site, the 178th site, the 214th site, the 219th site, the 220th site or the 220th site; the combined mutation adopts two modes: one mode is that a single-point mutation DHDR mutant is taken as a parent, mutation is carried out, and a combined mutation mutant I is obtained; and 2, taking the DHDR combined mutation mutant I as a parent, and performing mutation to obtain a combined mutation mutant II. The DHDR mutant disclosed by the invention is used for synthesizing (3S, 4R)-tetrahydrodaidzein by reducing dihydrodaidzein, and asymmetric reduction with high activity and high stereoselectivity can be realized.

The present invention relates to a process for producing a chiral aromatic amine, said process comprising the steps of: a) providing an aromatic imine, and b) contacting said aromatic imine from step a) with an imine reductase, thereby stereoselectively reducing said aromatic imine to a chiral aromatic amine with said imine reductase; and to novel imine reductases capable of catalytic reactions.

The present disclosure relates to recombinant microorganisms, such as bacteria, e.g., E. coli, engineered to have improved flux and / or yield of the DXP pathway. Cells can be engineered to comprise nucleotide sequences comprising dxs polypeptides (EC 2.2.1.7); dxr polypeptides (EC 1.1.1.267); ispD polypeptides (EC 2.7.7.60); ispE polypeptides (EC 2.7.1.148); ispF polypeptides (EC 4.6.1.12); ispG polypeptides (EC 1.17.7.1 or EC 1.17.7.3); ispH polypeptides (EC 1.17.7.4); and idi polypeptides (EC 5.3.3.2). Cells can be further engineered to express redox polypeptides, e.g., ferredoxins, flavodoxins, and / or flavodoxin / ferredoxin--NADP reductases (EC 1.19.1.1 or EC 1.18.1.2), e.g., to increase the activity of the ispG and / or ispH polypeptides. Some or all of the nucleotide sequences can be heterologous to the cells. The recombinant microorganisms can be used in methods of isoprenoid production.

The invention discloses a method for catalytic synthesis of malonic acid by using whole cells, and belongs to the field of bioengineering. According to the invention, escherichia coli BL21 (DE3) is taken as a host, an aspartic acid ammonia lyase gene aspA and an aspartic acid-alpha-decarboxylase gene panD, a succinate semialdehyde dehydrogenase gene yneI and a beta-alanine pyruvate transaminase gene, a beta-alanine pyruvate transaminase gene bauA and a malonyl-coenzyme A reductase gene mcr-C are expressed in modules, and recombinant escherichia coli is constructed. The malonic acid is produced by adopting a whole-cell catalysis method, so that the production period is shortened, the production efficiency of the malonic acid is improved, and the yield of the malonic acid can reach 51.0 g / L after the malonic acid is catalyzed for 24 hours.

The invention provides an engineered ketoreductase mutant, which comprises a polypeptide sequence, a gene sequence, a recombinant expression vector containing the gene, an engineering strain, a protein expression method of the engineering strain, and a reaction process for selectively synthesizing (R)-5, 7-difluorochroman-4-ol by using the engineered ketoreductase mutant.

The invention discloses a heme oxygenase mutant and application of the heme oxygenase mutant in preparation of bilirubin through multienzyme coupling, and belongs to the field of biological catalysis engineering. The forward mutant of the heme oxygenase is obtained through a directed evolution technology, and heme can be efficiently catalyzed to synthesize biliverdin; the bilirubin is synthesized by further introducing biliverdin reductase, coupling cytochrome P450 reductase and formate dehydrogenase, and coupling the two core enzymes with foreign assistant enzymes to construct recombinant escherichia coli to catalyze heme to synthesize bilirubin. In the system, NADPH does not need to be exogenously added, the reaction time is about 4 h, the yield of bilirubin reaches 2.10 g / L, accumulation of intermediate products is not observed in the catalysis process, and the highest yield and the shortest reaction time in bilirubin synthesis reported at home and abroad at present are achieved. Compared with the traditional pig bile extraction, the method disclosed by the invention breaks through the resource limitation, is simple to operate and high in yield, and realizes efficient and green synthesis of bilirubin.

The invention discloses a production method of fucosylated oligosaccharide. The core of the method is as follows: the GDP-D-rhamnose is converted into the GDP-L-fucose by using the GDP-D-rhamnose-3, 5-epimerase; the GDP-D-rhamnose is obtained by converting the GDP-D-mannose-4, 6-dehydratase and the GDP-4-keto-6-deoxy D-mannose reductase by taking the GDP-D-mannose as a substrate, and the GDP-D-rhamnose is obtained by taking the GDP-D-mannose as a substrate. The invention relates to a method for producing a fucosylated oligosaccharide, which is characterized in that a GDP-D-mannose-4, 6-dehydratase, a GDP-4-keto-6-deoxyD-mannose reductase, a GDP-D-rhamnose-3, 5-epimerase and a fucosyltransferase are expressed in a genetically modified host cell, and the fucosylated oligosaccharide can be produced by the way mentioned above.

There is disclosed a mutant, non-naturally occurring or transgenic tobacco plant or part thereof having modulated expression or activity of lysine ketoglutarate reductase (LKR) comprising, consisting of, or consisting essentially of: (i) a polynucleotide, the present invention relates to a protein comprising, consisting or consisting essentially of a sequence having at least 88% sequence identity to SEQ ID NO: 1 (NtLKR-S) and / or a sequence having at least 86% sequence identity to SEQ ID NO: 3 (NtLKR-T); (ii) a polypeptide encoded by the polynucleotide set forth in (i); (iii) a polypeptide comprising, consisting or consisting essentially of a sequence having at least 89% sequence identity to SEQ ID NO: 2 (NtLKR-S) and / or a sequence having at least 88% sequence identity to SEQ ID NO: 4 (NtLKR-T); or (iv) a construct, vector or expression vector comprising the isolated polynucleotide set forth in (i) wherein the plant or part thereof comprises at least one modification as compared to a control plant or part thereof wherein the expression of the polynucleotide or the activity of the polypeptide is not modified, the at least one modification is capable of modulating: (a) expression of the polynucleotide in the plant or part thereof; or (b) the activity of the polypeptide in the plant or part thereof.

The invention discloses a progesterone 5beta-reductase mutant and application thereof in synthesis of 5beta-dihydrosteroid, relates to the technical field of biology, and in particular relates to application of gene mining and engineering modification of progesterone 5beta-reductase derived from bacteria in asymmetric synthesis of 5beta-dihydrosteroid. According to the progesterone 5 beta-reductase LpP5beta R disclosed by the invention, the progesterone 5 beta-reductase LpP5beta R from bacteria psoromais is obtained by a gene mining method, and the progesterone 5 beta-reductase LpP5beta R can be subjected to soluble expression in escherichia coli. The mutant obtained by mutating the wild type LpP5beta R has obviously improved catalytic ability on progesterone, and can efficiently catalyze delta 4-3-carbonyl steroids such as hydrocortisone, dinorcitol and the like to generate corresponding 5beta-dihydrosteroids. According to the invention, the technical bottlenecks of poor heterologous expression and low catalytic efficiency of plant-derived progesterone 5beta-reductase are solved, and a potential biocatalyst is provided for green manufacturing of 5beta-dihydrosteroid drugs.

The invention relates to a method for preparing (S)-chiral amine through asymmetric catalysis of imine reductase, and belongs to the technical field of biological pharmacy. Comprising the following steps that imine reductase catalyzes imine to be subjected to a reductive hydrogenation reaction under the conditions of coenzyme and a coenzyme regeneration system, and the (S)-chiral amine compound is generated. According to the method, imine reductase is utilized to catalyze imine asymmetric reduction to prepare the innovative drug (S)-chiral amine compound, and compared with a traditional chemical synthesis method, the method has the advantages of being high in reaction efficiency, good in stereoselectivity, mild in reaction condition, environmentally friendly and the like, and is a method meeting the green production requirement.

The invention discloses an enzyme catalysis preparation method of a milobalin intermediate. According to the method, racemization 3-ethyl bicyclo [3.2. 0] hept-3-ene-6-ketone is used as a substrate, specific ketoreductase is used as a catalyst, (1S, 5R)-3-ethyl bicyclo [3.2. 0] hept-3-ene-6-ketone is selectively reduced into (1S, 5R)-3-ethyl bicyclo [3.2. 0] hept-3-ene-6-alcohol, and a target compound milobalin intermediate (1R, 5S)-3-ethyl bicyclo [3.2. 0] hept-3-ene-6-ketone is reserved. According to the method disclosed by the invention, only one enzyme is used, the concentration of a conversion substrate can reach 200g / L, the yield can reach 47.4%, the ee value can reach 99.8%, the operation is simple, the resolution efficiency is high, the selectivity is good, and the method can be used for industrial production.

The invention discloses a Yarrowia lipolytica engineering bacterium for producing astaxanthin in a peroxisome compartment and application of the Yarrowia lipolytica engineering bacterium. A complete astaxanthin synthesis path is expressed in host bacteria by recombinant Yarrowia lipolytica; comprising geranyl geranyl diphosphate synthase CrtE, phytoene synthetase / lycopene cyclase CrtYB, phytoene desaturase CrtI, 3-hydroxy-3-methylglutaryl CoA reductase tHMGR, beta-carotene ketolase CrtW and beta-carotene hydroxylase CrtZ, and the enzymes are relocated into peroxisome, so that the enzyme activity of the peroxisome is improved, and the enzyme activity of the peroxisome is improved. The number of expression hemoglobin genes VHb and peroxisome is increased, the surface area of the hemoglobin genes VHb is enlarged, and one or more of enzymes PEX11, PEX19, DNM1 or PHO85 for prolonging the service life of the hemoglobin genes VHb and peroxisome is / are obtained; the astaxanthin production performance of the recombinant strain is verified based on cytoplasm engineering and subcellular organelle compartment engineering, and the astaxanthin production capacity of the yarrowia lipolytica is further improved.

A method for reducing a substrate selected from 2-methyl-5-nitropyridine and methyl 4-(2-fluoro-3-nitrobenzyl)piperazine-1-carboxylate is provided catalysed by a nitroreductase and a disproportionation agent.

The invention provides immobilized ketoreductase, an immobilization method and application of the immobilized ketoreductase in preparation of a key intermediate of bentonatate, the amino acid sequence of the ketoreductase is as shown in SEQ ID NO.2, preferably a ketoreductase mutant is as shown in SEQ ID NO.16, and the immobilized ketoreductase comprises resin and corresponding ketoreductase. The ketoreductase mutant and the immobilized ketoreductase prepared from the ketoreductase mutant are very high in enzyme activity during ketoreduction reaction, and when the ketoreductase mutant is used for preparing a key intermediate of bentonamate, a solvent is pure water, so that the ketoreductase mutant and the immobilized ketoreductase prepared from the ketoreductase mutant are environment-friendly. The immobilized transaminase is high in conversion rate, good in stereoselectivity, good in stability, reusable and simpler to operate, so that the production cost is reduced, and industrial production is facilitated.

The present application provides a kind of Radix Polygoni Multiflori active ingredient separation and purification process, belong to plant extraction technical field.The present application separation and purification process includes: Radix Polygoni Multiflori ethanol ultrasonic extraction, activated carbon adsorption, HPD-100 macroporous resin purification, membrane filtration, ultrafiltration, reduced pressure concentration and crystallization step, can efficiently extract Radix Polygoni Multiflori extract with high 5 alpha-reductase inhibitory activity.After compounding with the extract of Euonymus alatus obtained by specific microbial fermentation, the anti-hair loss effect can be significantly enhanced.The in vitro experimental results show that the Radix Polygoni Multiflori extract and the extract of Euonymus alatus both have significant 5 alpha-reductase inhibitory activity.The present application can effectively promote hair growth and improve hair loss, and the effect is close to the positive control drug.The present application process is stable, high in purity, strong in activity, suitable for industrial production, and has good market application prospect.

A method for catalytically synthesizing a single chiral 2-aryl-substituted nitrogen heterocyclic derivative by means of an imine reductase, and a use thereof. The method comprises: under the catalytic action of an imine reductase, converting a compound represented by formula II into a single chiral compound represented by formula I. In formulas I and II, each R is independently selected from hydrogen, fluorine, chlorine, bromine, C1-C6 alkyl, halogenated C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, cyano, nitro, or carboxyl; X is selected from -CH2- or -O-; and n is an integer selected between 0 and 6. The imine reductase is selected from any one of or a combination of at least two of the following amino acid sequences: (1) an amino acid sequence as shown in any one of SEQ ID NOs: 2, 4, 6, 7, or 10; and (2) an amino acid sequence having imine reductase activity and having at least 90% identity to the sequence as shown in any one of SEQ ID NOs: 2, 4, 6, 7, or 10.

Heterologous synthesis of a nitrogen-fixing system in a non-diazotrophic organism is a long-sought-after goal because of the significance of nitrogenase for areas related to agronomy, energy and environment. Two simplified nitrogenase analogs from Azotobacter vinelandii in Escherichia coli were successfully heterologously synthesized. One analog comprise the reductase component (NifH) and, in place of the catalytic component (NifDK), a cofactor maturase (NifEN); whereas the other analog comprised only NifEN. Metal, EPR and activity analyses verified the cluster composition and functional competence of the heterologously expressed NifH and NifEN. Moreover, NMR, nanoSIMS and growth experiments illustrated the abilities of both systems to mimic the nitrogenase in reducing N2 and incorporating the reduced N into the cellular mass. These results firmly established NifEN / NifH or NifEN as a simplified nitrogenase analog that could be engineered or expanded to facilitate future transgenic expression of nitrogenase and development of nitrogenase-based biotechnological applications.

The invention discloses a carboxylic acid reductase DeCAR recombinant plasmid and application of the carboxylic acid reductase DeCAR recombinant plasmid in preparation of aldehyde spices, belongs to the technical field of bioengineering, and further provides a recombinant bacterium containing the recombinant plasmid, and the carboxylic acid reductase DeCAR is further produced by using the recombinant bacterium, or the recombinant bacterium is directly applied to production of the aldehyde spices. Compared with other biocatalysts for preparing aldehydes, the carboxylic acid reductase and the recombinant bacteria thereof provided by the invention have the advantages of wide substrate spectrum, high catalytic activity, strong substrate specificity and the like, and have wide application prospects in industrial application.