16 results about "Carbonyl Reductase" patented technology

The present invention provides a carbonyl reductase mutant, preparation method and use thereof, and a preparation method of ethyl (R)-6-hydroxy-8-chlorooctanoate. The carbonyl reductase mutant is a carbonyl reductase with amino acid mutation; the carbonyl reductase comprises an amino acid sequence as set forth in SEQ ID NO: 2; the amino acid mutation includes E101V, F214R or E101V / F214R. In the present invention, by introducing mutations on the basis of the original carbonyl reductase sequence, the enzyme activity is improved, the stereoselectivity is improved, and ethyl (R)-6-hydroxy-8-chlorooctanoate can be obtained with high yield and high purity under relatively mild conditions, which reduces the production cost and is suitable for industrial production.

The application discloses a carbonyl reductase mutant, a combined enzyme and application of the combined enzyme in synthesis of S-configuration boselike, and belongs to the technical field of biological catalysis and pharmacy. The amino acid sequence of the wild-type carbonyl reductase mutant provided by the application is shown as SEQ ID NO:1, and the mutant is obtained by mutation of the wild-type carbonyl reductase, and the mutation mode is that F at the 147th position is mutated into L, V at the 163rd position is mutated into Y, L at the 176th position is mutated into E, and P at the 200th position is mutated into R. By using the mutant, a catalytic system is formed by combining the mutant with formic acid dehydrogenase (FDH), and optical pure S-configuration boselike can be efficiently and highly selectively prepared under high substrate concentration of 250-320 g / L, separation and purification problems caused by generation of a large amount of soluble by-products in catalysis of other combined enzymes are avoided, a post-treatment process is obviously simplified, and production cost and environmental burden are reduced.

The invention discloses a carbonyl reductase immobilization process and application of the carbonyl reductase immobilization process in synthesis of an ezetimibe intermediate, immobilized enzyme particles are prepared through activation of epoxy resin and loading of carbonyl reductase and glucose dehydrogenase, and then ZT-4 is catalyzed by the immobilized enzyme particles to chirally synthesize a ZT-5 product. The method disclosed by the invention has the beneficial effects that the enzyme can be repeatedly utilized through an enzyme immobilization process, so that the use cost of the enzyme is effectively reduced, and the use amount of expensive coenzyme is reduced through a glucose regeneration method, so that the production cost is effectively saved; according to the method, the treatment step of free enzyme is omitted by separating the immobilized enzyme, so that the reaction short batch interval can be effectively shortened, the generation of waste gas in the reaction post-treatment is reduced, the environmental protection property of the reaction is ensured, the production requirement of green industrialization is met, and the method is also suitable for large-scale production.

This invention discloses a chemical-biological fusion method for cholesterol synthesis. Using plant-derived 21-hydroxy-20-methylpregn-4-en-3-one, also known as bis(2-hydroxy-2-yl)-3-one (BA), as a raw material, cholesterol is synthesized through a four-step reaction involving sulfonation, acetylation, enzymatic hydrolysis and reduction, and Grignard coupling. Addressing the drawbacks of traditional animal-derived cholesterol, which carries unpredictable risks of pathogenic bacteria and viruses, this invention uses plant-derived steroidal raw material BA to synthesize cholesterol. This method not only offers high safety, avoiding the risk of pathogenic bacteria and viral infections, but also utilizes the tandem catalysis of esterase and carbonyl reductase during synthesis, along with coenzyme regeneration, to obtain key intermediates with high stereoselectivity and in a safe and clean manner. This significantly shortens the cholesterol synthesis route, increases efficiency, simplifies operation, minimizes solvent recycling losses, reduces waste, and allows for mild reaction conditions. This method is safe, economical, and suitable for industrial production.

The invention discloses a method for separating and preparing (R)-citronellal and co-producing (S)-citronellal, which takes raw materials containing (R)-citronellal and (S)-citronellal as substrates and carbonyl reductase and formate dehydrogenase as catalysts. According to the invention, co-production of (R)-citronellal and (S)-citronellal can be realized, and the (R)-citronellal and (S)-citronellal can be applied to the field of spices,

The invention belongs to the technical field of biological catalysis, and particularly relates to a carbonyl reductase mutant and application of the carbonyl reductase mutant in synthesis of an R-lipoic acid chiral intermediate (R)-6-hydroxy-8-chloro ethyl caprylate. According to the method, an unseparated and purified 6-oxy-8-chloro ethyl caprylate stock solution is used as a substrate, carbonyl reductase mutant coupled glucose dehydrogenase BtGDH is used as a catalyst, and the (R)-6-hydroxy-8-chloro ethyl caprylate is asymmetrically synthesized under mild conditions (20-40 DEG C, pH is 6-9). The (R)-6-hydroxy-8-chloro ethyl caprylate enzyme can be used for completely converting 500 g / L of 6-oxy-8-chloro ethyl caprylate stock solution within 8 hours to synthesize optically pure (R)-6-hydroxy-8-chloro ethyl caprylate. According to the method, the substrate does not need to be pretreated (solvent removal), the synthesis yield can reach 87%, and the method has the advantages of being high in space time yield, simple in process, green, environmentally friendly and the like.

The invention relates to an enzyme immobilization carrier and application thereof, and belongs to the technical field of enzyme immobilization, the enzyme immobilization carrier comprises a magnetic Fe3O4-SiO2 nano core with a particle size of 20-50nm; the magnetic core is coated with the thermo-sensitive poly (N-isopropylacrylamide) hydrogel layer; the aminated mesoporous silica is grafted on the surface of the hydrogel layer, the aperture is 5-10nm, and the specific surface area is greater than or equal to 800m < 2 > / g; a carbonyl reductase GJ3401 covalently immobilized on the mesoporous silica; the coenzyme NADP and the N-isopropylacrylamide hydrogel layer are crosslinked on the surface of the carrier through a PEG chain, sodium alginate is added into the hydrogel layer, the mass ratio of the sodium alginate is 1-3%, and the overall particle size of the carrier is 150-200 [mu] m; the method integrally has the advantages of co-immobilization of the carbonyl reductase and the coenzyme, high enzyme activity stability, efficient cyclic utilization of the coenzyme and easiness in separation and recovery.

The invention relates to the technical field of synthesis of (R)-lipoic acid precursors, in particular to an immobilized enzyme carrier PVA-DMC-coated Al2O3, an immobilized enzyme, a preparation method of the immobilized enzyme carrier PVA-DMC-coated Al2O3 and the immobilized enzyme and a preparation method of (R)-8-chloro-6-hydroxy ethyl caprylate. The immobilized enzyme comprises a carrier as well as carbonyl reductase and glucose dehydrogenase which are co-immobilized on the carrier, and the carrier is an immobilized enzyme carrier PVA-DMC coated Al2O3; the immobilized enzyme carrier PVA-DMC coated Al2O3 comprises polyvinyl alcohol and a reinforcing agent loaded on the polyvinyl alcohol, and the reinforcing agent contains aluminum oxide and dimethyl carbonate. When the immobilized enzyme provided by the invention is used in the process of catalytically preparing (R)-8-chloro-6-hydroxy ethyl caprylate, high-concentration conversion of a substrate can be realized, the conversion rate can be increased, meanwhile, the swelling phenomenon of a carrier can be greatly reduced, the cycle use frequency of the carrier is increased, the cycle use frequency is not less than 24 times, and the conversion rate is not less than 95%.

The present invention belongs to the technical field of biocatalysts. Disclosed are an alkene reductase mutant and the use thereof in the catalytic preparation of an intermediate for the synthesis of brivaracetam. The amino acid sequence of the alkene reductase mutant of the present invention is as shown in SEQ ID NO. 1. A compound enzyme composed of the mutant, carbonyl reductase K1 and glucose dehydrogenase can, with the compound 5-hydroxy-4-n-propyl-2-furanone as a substrate, reduce double bonds and remove hydroxyl, so as to obtain the compound (R)-4-propyl dihydrofuran-2(3H)-one; and the ee value of the (R)-4-propyl dihydrofuran-2(3H)-one obtained by means of catalysis reaches 93.0%, and (R)-4-propyl dihydrofuran-2(3H)-one having an ee value of greater than 99.8% can be further obtained by means of the crystallization of diastereoisomers.

The present application relates to the technical field of biology, and particularly relates to a complex enzyme, a co-immobilized enzyme and application thereof. The present application provides a complex enzyme, a co-immobilized enzyme and application thereof, and a preparation method of Bose factor, wherein the preparation method takes 1-C-(beta-D-pyranosyl) acetone as an initial substrate, selectively synthesizes high-purity S-Bose factor in one step under the action of a carbonyl reductase, introduces formic acid dehydrogenase to regenerate an NADH system, takes NAD and sodium formate as substrates, cyclically generates NADH and carbon dioxide, reduces the accumulation of intermediate products, and finally stably realizes the generation of 380 g / L products, the reaction conversion rate is greater than 99%, the ee value is 100%, the immobilized enzyme is used for 15 times, the enzyme activity retention rate is above 85%, and compared with the free enzyme, the cost is reduced by more than 5 times.

The invention discloses an immobilized enzyme for preparing a ticagrelor intermediate and a preparation method of the immobilized enzyme. The immobilized enzyme comprises a carbonyl reductase mutant and an immobilized carrier, and the amino acid sequence of the carbonyl reductase mutant is as shown in SEQ ID NO: 1; the immobilized carrier is selected from the group consisting of LX1000EA, LX1000IDA, LX1000HA and LX1000ME, and the immobilized carrier is selected from the group consisting of LX1000EA, LX1000IDA and LX1000ME. The immobilized enzyme provided by the invention is used as a biocatalyst for biocatalytic preparation of a ticagrelor intermediate (S)-2-chloro-1-(3, 4-difluorophenyl) ethanol, the product yield and purity are high, the repeated utilization rate of the enzyme is high, and the separation and purification process is simple.

The application discloses a continuous enzymatic preparation method and a continuous production process of ursodeoxycholic acid, and belongs to the technical field of biological catalysis and drug synthesis. The core of the method is as follows: a carrier with an isoelectric point greater than 9.5 is used to covalently fix a carbonyl reductase and a coenzyme regeneration enzyme, the surface of the carrier is positively charged under the reaction condition, the large-molecule coenzyme with negative electricity is locally concentrated near the enzyme activity center through electrostatic adsorption, the oil-in-water microemulsion reaction system is constructed to efficiently solubilize the substrate, the dynamic pH gradient regulation is implemented in the continuous flow reaction bed to cooperatively optimize the double-enzyme activity, the online membrane separation and extraction unit is integrated to realize the efficient interception and reflux of the unreacted coenzyme and the immediate removal of the product, and the process realizes the efficient and high-selectivity continuous synthesis of the UDCA, the coenzyme utilization efficiency and the production space-time yield are significantly improved, the catalyst operation stability is good, and the process is suitable for the large-scale production of the medical-grade UDCA.

This invention discloses a carbonyl reductase mutant and its application in the synthesis of S-configuration Bosein, belonging to the field of biocatalytic synthesis technology. The carbonyl reductase mutant is based on the carbonyl reductase with the amino acid sequence shown in SEQ ID No. 1, including mutation types F132A, V162A, F132A-N9E, F132A-K36R, and F132A-N9E-K36R. This invention also discloses the nucleic acid molecule encoding the mutant, the expression vector, the genetically engineered strain, and a method for synthesizing S-configuration Bosein using this mutant in combination with isopropanol dehydrogenase. This carbonyl reductase mutant exhibits significantly enhanced enzyme activity and can be adapted to low-cost coenzymes. When synergistically catalyzed with isopropanol dehydrogenase, it can efficiently reduce β-pyruvate xyloside to S-configuration Bosein, with high conversion rate, short reaction time, and high product purity, significantly reducing the production cost of S-configuration Bosein and showing promising industrial application prospects.

The invention discloses a carbonyl reductase mutant and application thereof in preparation of chiral alcohol, and relates to the technical field of biological pharmacy and biochemical engineering. The carbonyl reductase mutant is obtained by mutating at least one site in SEQ ID NO: 2: the 119th site, the 180th site, the 183rd site, the 185th site, the 219th site, the 227th site and the 236th site. The invention further provides application of the carbonyl reductase mutant in catalyzing asymmetric reduction reaction of carbonyl compounds to prepare chiral alcohol and a preparation method of (R)-4-chromogen alkanol compounds. The carbonyl reductase mutant is constructed, the (R)-4-chromogen alkanol compound is prepared through a biological preparation way by adopting the carbonyl reductase mutant as a catalyst, and the method is environment-friendly, simple and convenient to operate, easy to industrially amplify, high in substrate concentration, wide in substrate universality and high in product yield, and has a very good industrial application prospect.

The application discloses a carbonyl reductase mutant, a preparation method and application thereof, and belongs to the technical field of enzyme engineering. A series of advantageous mutants are obtained by the method of directional evolution in the field of enzyme engineering on the basis of the carbonyl reductase BcKRED from Bacillus cereus. The obtained carbonyl reductase advantageous mutant has high catalytic activity, thermal stability and reversed chiral selectivity compared with the wild-type carbonyl reductase. The application provides a green and efficient way for industrial production of chiral alcohol by using the carbonyl reductase as a biological catalyst, and has a wide application prospect.

The invention discloses a carbonyl reductase mutant with high organic solvent tolerance and application thereof, the carbonyl reductase ZmCR mutant constructed by the invention has significantly improved temperature and organic dissolution tolerance, and significantly improved concentration and efficiency of a catalytic substrate: the catalytic efficiency of the mutant ZmCR-M3 is significantly improved; when the strain acts on 1000 mM of 1-C-(beta-D-xylopyranosyl)-acetone, the conversion rate of (S)-1-C-(beta-D-xylopyranosyl)-propanol generated within 24 hours is far higher than that of ZmCR (Zinc-CR). According to the carbonyl reductase mutant with improved organic solvent tolerance, the tolerance to solvents isopropanol and acetone is improved, the industrial application range is expanded, and the carbonyl reductase mutant has a great application prospect.