595 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.

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 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 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.

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.

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.

A recombinant yeast cell functionally expressing: a) a nucleic acid sequence encoding an enzyme having NADH-dependent nitrate reductase activity and / or a nucleic acid sequence encoding an enzyme having NADH-dependent nitrite reductase activity; and b) a nucleic acid sequence encoding a protein having transketolase activity (EC 2.2.1.1), wherein the expression of the nucleic acid sequence encoding the protein having transketolase activity is under control of a promoter (the “TKL promoter”), which TKL promoter has an anaerobic / aerobic expression ratio for the transketolase of 2 or more.

The invention discloses a dihydrodaidzein reductase, a combinatorial enzyme and an application of the dihydrodaidzein reductase and the combinatorial enzyme in synthesis of S-equol. The dihydrodaidzein reductase is a mutant which takes the dihydrodaidzein reductase AeDHDR as a starting sequence and has a corresponding mutation site, wherein the dihydrodaidzein reductase AeDHDR is derived from AdleCRzia equaliforns, and the amino acid sequence of the dihydrodaidzein reductase AeDHDR is shown as SEQ ID NO: 2. The invention also discloses a coding gene, a carrier and a bioengineering bacterium of the dihydrodaidzein reductase mutant and a method for synthesizing S-equol. The method has the advantages of high enzyme activity and high S-equol yield.

The invention belongs to the technical field of biological engineering, and provides curcumin / dihydrocurcumin reductase MaCurA, a coding gene and application. The reductase can efficiently catalyze 1, 7-bis (4-hydroxy-3-methoxyphenyl)-1, 6-heptadiene-3, 5-diketone to synthesize tetrahydrocurcumin, an efficient solution is provided for industrial preparation of tetrahydrocurcumin, and experiments prove that the recombinase can catalyze the synthesis of tetrahydrocurcumin in the catalysis of 1, 7-bis (4-hydroxy-3-methoxyphenyl)-1, 6-heptadiene-3, 5-diketone. In a reaction system for preparing tetrahydrocurcumin from 2, 5-diketone, actually measured enzyme activity is stabilized at 3.38 U / mg level, and compared with a traditional chemical synthesis method, the production efficiency is remarkably improved.

The invention discloses a sclareol synthase mutant, a recombinant genetic engineering strain and application of the sclareol synthase mutant, and belongs to the technical field of biology. The sclareol synthase is modified through semi-rational design, the obtained sclareol synthase mutant has good catalytic activity, and the yield of sclareol can be remarkably increased; furthermore, the method is simple. The method comprises the following steps: integrating an HMG1 gene for coding HMG-CoA reductase, an IDI1 gene for coding isopentenyl pyrophosphate isomerase, an SsGGPPS gene for coding geranyl-geranyl pyrophosphate synthase, an SsLPPS gene for coding lybaidan diglycol diphosphate synthase, and an SsGGPPS gene for coding lybaidan diglycol diphosphate synthase; a recombinant genetic engineering strain obtained by the nucleic acid molecule for coding the sclareol synthase mutant can produce sclareol at high yield by taking glucose as a substrate, and has a remarkable industrial application value.

The invention discloses an oxidoreductase mutant and application thereof, and belongs to the field of enzyme engineering. The invention provides a recombinant oxidoreductase HbKR mutant which is used for catalytically producing (R)-decalactone. The oxidoreductase HbKR mutant disclosed by the invention realizes soluble expression in Escherichia coli BL21 (DE3), and the oxidation activity of the oxidoreductase HbKR mutant is improved by 4.61 times compared with that of a parent enzyme. An oxidoreductase HbKR mutant and lactone hydrolase Rpa3624 derived from Rhodopseudomonas palustris are combined for use, reaction is performed for 0.5 h under the conditions that the temperature is 35 DEG C and the pH value is 10.5, compared with a wild type, the yield of (R)-delta-decalactone is increased to 48.2% from 42.3%, and the stereoselectivity is increased to 98.7% from 94.2%.

The invention provides a method for preparing (R)-4-chloro-3-hydroxybutyric acid ethyl ester ((R)-CHBE) through enzyme catalysis, which comprises the following steps: isopropanol is used as a hydrogen donor, a ketoreductase mutant is used for carrying out catalytic reduction reaction of COBE to prepare (R)-CHBE, and the ketoreductase mutant has amino acid mutation at the 83rd site on the basis of an amino acid sequence shown as SEQ ID NO.1. The invention also provides a preparation method of (R)-4-chloro-3-hydroxybutyric acid ethyl ester ((R)-CHBE). The thermal stability, namely heat resistance, of the ketoreductase mutant is improved, and by combining with the use of a rectifying tower, the method disclosed by the invention improves the utilization rate of isopropanol and can discharge a byproduct acetone in time, so that the substrate conversion rate and the product concentration are improved. The invention also provides an enzyme catalysis device used in the method.

The invention relates to the field of gene engineering, in particular to a daidzein reductase mutant and application of the daidzein reductase mutant in preparation of equol, and the daidzein reductase mutant is prepared by performing one or more of the following mutations on the basis of a wild type: V263F, P107A, H351R, G35L, H171A, A267Q, H351Q, M355Q, H351P, G31V, V176A, H351Y, N64K, V137A, H351W, D63A, W257A and C174N. The catalytic activity of the mutant obtained through precise mutation is remarkably improved under the feedback inhibition condition, especially when high-concentration equol exists, the mutant can still keep high relative activity, and continuity and high efficiency of the biosynthesis process are ensured. When the mutant is applied to biosynthesis of equol, the production titer is remarkably improved, the yield can be improved by about 1.2 times compared with that of a wild type, and the economic benefit and the yield of biosynthesis are directly improved.