145 results about "Carbonyl Reductase" patented technology

The invention discloses a carbonyl reductase mutant and a method for catalytically synthesizing high-concentration glassine by using the carbonyl reductase mutant, and belongs to the field of biological catalysis. According to the invention, a one-pot method is adopted to catalyze beta-acetone xyloside to synthesize high-concentration boine, carbonyl reductase CnMDR coded enzyme and rational design of a substrate pocket key are combined with multiple sites, beneficial mutations are combined, dominant mutant enzyme recombinant bacteria are used as a catalyst, beta-acetone xyloside is used as a substrate, and formate dehydrogenase variants and NAD + kinase are introduced at the same time, so that the high-concentration boine is synthesized. The NADP < + >-NADPH coenzyme is driven to circulate, the yield of the synthesized boine is 200g / L, and the conversion rate is gt; the ee value of the product is gt; the invention provides a one-pot efficient low-cost method for synthesizing the boine, only a trace amount (0.3 mM) of coenzyme needs to be added, byproducts in a reaction system are easy to separate, and great convenience is provided for industrial large-scale production.

The invention provides a carbonyl reductase mutant. The carbonyl reductase mutant is obtained by performing single-point mutation or multi-point combined mutation on the 17th site, the 40th site and the 64th site of an amino acid sequence as shown in SEQ ID NO.1. The invention also provides a coding gene, a recombinant vector containing the coding gene, a co-expression engineering bacterium and an application of the co-expression engineering bacterium. Compared with a wild type enzyme, the carbonyl reductase mutant has high activity and high stereoselectivity, the enzyme activity can reach more than two times that of the wild type enzyme, the catalytic efficiency on a substrate precursor ketone is remarkably improved, the yield of a statin drug intermediate and chiral alcohol synthesized by an enzyme method of the carbonyl reductase mutant is remarkably improved, and the yield of a rosuvastatin intermediate (3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R) is remarkably improved. The highest yields of the (3R, 5R)-6-chloro-3, 5-dihydroxyhexanoic acid tert-butyl ester and the atorvastatin intermediate (3R, 5R)-6-cyano-3, 5-dihydroxyhexanoic acid tert-butyl ester reach 99% and 97.6% respectively, e.e. Is larger than 99%, and the method has high industrial application value.

The invention discloses a carbonyl reductase, a mutant and an application of the carbonyl reductase in synthesis of a chiral alcohol compound. The carbonyl reductase mutant is obtained by performing single-site or multi-site mutation on the 96th site, the 145th site, the 202nd site or the 206th site of the amino acid sequence of the carbonyl reductase LbCR derived from lactobacillus brevis as shown in SEQ ID No.2. The activity and stability of the carbonyl reductase LbCR mutant are obviously improved, and the carbonyl reductase LbCR mutant has a wide substrate spectrum. Compared with a wild type, the mutant LbCRM4 has the advantages that the enzyme activity is improved by 86.3 times, the substrate conversion rate is greater than 99%, the e.e.p value of the product is always kept at 99.5% or above, the space time yield reaches 1288.9 g / L / d, and the half-life period at 40 DEG C is 101.4 h and is improved by 83.5 times. The LbCR mutant has an industrial application prospect.

The invention discloses a high-performance carbonyl reductase mutant and application thereof in synthesis of a series of chiral alcohols. The high-performance carbonyl reductase mutant is obtained by performing the following mutations on an amino acid sequence as shown in SEQ ID NO.2: the 69th alanine mutates into any one of lysine, leucine, aspartic acid or asparagine, and the 69th alanine mutates into any one of lysine, leucine, aspartic acid or asparagine; and / or the 100th glycine is mutated into any one of glutamic acid, serine, lysine or asparagine. Compared with a wild type enzyme, the thermal stability and the catalytic activity of the obtained mutant are remarkably improved, particularly, single point mutants Mut-A69N, Mut-G100E and Mut-G100N and a combined mutant Mut-A69N-G100N show excellent relative enzyme activity, thermal stability and substrate tolerance. The mutant has high catalytic activity and stability to various chiral alcohols, and the contradiction that the traditional carbonyl reductase is easy to inactivate at high temperature and the catalytic efficiency is difficult to achieve at the same time is solved. The invention provides an efficient and stable enzyme catalysis tool for green biological preparation of chiral drug intermediates.

The invention discloses a carbonyl reductase mutant as well as a preparation method and application thereof, and belongs to the technical field of enzyme engineering. According to the invention, on the basis of carbonyl reductase BcKRED from Bacillus cereus, a series of dominant mutants are obtained through a method of directed evolution in the field of enzyme engineering. The carbonyl reductase dominant mutant obtained by the invention has high catalytic activity, thermal stability and chiral selectivity reversed with wild-type carbonyl reductase. A green and efficient way is provided for industrial production of chiral alcohol by taking carbonyl reductase as a biocatalyst, and the method has a wide application prospect.

The invention relates to the technical field of biology, and discloses a carbonyl reductase mutant and application thereof in enzymatic synthesis of chiral alcohol. The amino acid sequence as shown in SEQ ID NO.2 is subjected to mutation that 36th lysine is mutated into any one of aspartic acid, threonine and valine, or / and 125th histidine is mutated into any one of lysine, serine and leucine, or 170th histidine is mutated into any one of lysine, cysteine and leucine. The carbonyl reductase mutants, especially single-site mutants Mut-T75K, Mut-T75D and Mut-E216M and a combined mutant Mut-T75K-E216M, with activity improved to different extents compared with that of a wild type are obtained, have excellent relative enzyme activity, thermal stability and substrate tolerance, have relatively high activity and stability for various chiral alcohols, and can be used for preparing the carbonyl reductase mutants. The catalyst has the advantages of high catalytic activity, high thermal stability, strong substrate tolerance, easiness in fermentation and the like.

The invention provides a carbonyl reductase mutant and application thereof in preparation of chiral alcohol compounds, and belongs to the technical field of gene engineering and enzyme catalysis. According to the invention, on the basis of the wild carbonyl reductase, the mutant with activity obviously superior to that of the wild carbonyl reductase is obtained through specific mutation, the selectivity and stability of the enzyme are improved, and the mutant has a wide application prospect in catalytic synthesis of chiral alcohol compounds.

The invention belongs to the technical field of perfume synthesis, and particularly relates to a method for efficiently preparing chiral oxo-ionol by an enzymic method, which comprises the following steps: putting a substrate containing oxo-ionone and an auxiliary substrate into a buffer solution, and stirring at room temperature until complete dissolution to obtain a substrate solution; the method comprises the following steps: dispersing carbonyl reductase ChKRED20 or a mutant thereof and coenzyme into a substrate solution, and carrying out heat preservation and stirring; and after the reaction is finished, adding water to dilute, extracting with ethyl acetate, washing with water, drying, and concentrating the ethyl acetate to obtain the chiral oxo-ionol. The enzyme catalysis method disclosed by the invention is mild in reaction condition and high in selectivity, and a target product can be obtained at high yield at room temperature and normal pressure. The coenzyme is regenerated by adopting a substrate coupling method, the use amount of the coenzyme is reduced, and a substrate coupling strategy that an auxiliary substrate is used as a substrate is adopted, so that a reaction system is simple, products are easy to separate, and the cost is reduced. According to the technical scheme, the conversion rate is greater than 99%, the ee value of the product reaches 99% or above, and the industrial application value is achieved.

The invention provides a carbonyl reductase mutant and an application of the carbonyl reductase mutant in synthesis of S-glassine. The mutant protein is a non-natural protein, and the mutant protein can significantly improve the activity of catalyzing the reduction of carbonyl in 1-C-(beta-D-xylopyranosyl)-acetone to generate S-configuration alcohol. When the activity-improved carbonyl reductase mutant strain is applied to the production process of S-configuration glassine, the carbonyl reduction capacity of the S-configuration glassine can be improved, 100-240 g / L of substrate can be completely converted, and the S-configuration glassine is generated.

The invention belongs to the technical field of biocatalysts, and discloses an alkene reductase mutant and application of the alkene reductase mutant in catalytic preparation of a brivaracetam synthesis intermediate. The amino acid sequence of the alkene reductase mutant is as shown in SEQ ID NO.1, a compound enzyme composed of the mutant, carbonyl reductase K1 and glucose dehydrogenase can take a compound 5-hydroxy-4-n-propyl-2-furanone as a substrate, double bonds are reduced, hydroxyl is removed, a compound (R)-4-propyl dihydrofuran-2 (3H)-ketone is obtained, and the compound (R)-4-propyl dihydrofuran-2 (3H)-ketone can be used for preparing the alkene reductase. The ee value of the (R)-4-propyl dihydrofuran-2 (3H)-ketone obtained through catalysis reaches 93.0%, the (R)-4-propyl dihydrofuran-2 (3H)-ketone with the ee value larger than 99.8% can be further obtained through diastereoisomer crystallization, and the method can be used for efficient and green preparation of the brivaracetam key intermediate (R)-4-propyl dihydrofuran-2 (3H)-ketone.

The invention discloses a carbonyl reductase mutant as well as a preparation method and application thereof, and belongs to the technical field of gene engineering and enzyme engineering. According to the invention, wild-type carbonyl reductase of Stachybotrys chlorohallonata is taken as an evolution template, and a plurality of mutants with catalytic activity, thermal stability and substrate spectrum obviously superior to those of the wild-type carbonyl reductase are obtained through an enzyme engineering technology. The carbonyl reductase mutant can catalyze various prochiral carbonyl compounds to obtain chiral alcohol with high stereoselectivity (ee is greater than 99%). The carbonyl reductase mutant provides a novel, efficient and environment-friendly technical route for industrial production of chiral alcohol products, and has very high industrial application potential.

The invention discloses a high-stability carbonyl reductase mutant and application of the high-stability carbonyl reductase mutant in chiral alcohol synthesis. An amino acid sequence as shown in SEQ ID NO.2 is mutated as follows: 20th leucine is replaced by any one of lysine, threonine, aspartic acid or isoleucine, and / or 55th leucine is replaced by any one of methionine, serine, lysine or isoleucine, and / or 55th leucine is replaced by any one of methionine, serine, lysine or isoleucine. Compared with a wild type enzyme, the stability of the obtained mutant is remarkably improved, particularly, single-point mutants Mut-L20I and Mut-L55I and a combined mutant Mut-L20I-L55I show excellent relative enzyme activity, thermal stability and substrate tolerance. The mutant has high catalytic activity and stability to various chiral alcohols, has the advantages of high catalytic efficiency, high thermal stability, strong substrate tolerance, easiness in fermentation production and the like, and is suitable for industrial application of enzymatic synthesis of chiral alcohols.

The invention relates to the technical field of NADPH regeneration and the technical field of antidepressant drug synthesis, and discloses a method for catalytically synthesizing (S)-3-chloro-1-phenyl-1-propanol by driving a WO3-based photoenzyme catalyst through infrared light, and the method comprises the following steps: step S1, preparing an oxygen-deficient WO3 nanosheet; s2, preparing a NaCBR carbonyl reductase (NaCBR carbonyl reductase); s3, mixing the oxygen-deficient WO3 nanosheet and NaCBR carbonyl reductase according to the mass ratio of (1: 0.5)-(1: 2), and stirring for 3-6 hours to obtain the photo-enzyme catalyst (WO3 / NaCBR). According to the method, artificial photosynthesis, namely a method for driving NADPH regeneration, is utilized, other materials are not additionally used as hydrogen transfer and electron transfer media, and the (S)-3-chloro-1-phenylpropanol is catalytically synthesized by utilizing infrared light to drive a photo-enzyme catalyst (WO3 / NaCBR). The method can greatly reduce the cost, is green and environment-friendly, has high catalytic efficiency and mild reaction conditions, is simple to operate, is easy to recover and recycle, and is suitable for large-scale industrial production and application.

The invention relates to the technical field of biology, and discloses a recombinant carbonyl reductase mutant and application thereof in synthesis of chiral alcohol. According to the invention, the 36th-site lysine is mutated into any one of aspartic acid, threonine and valine, or / and the 125th-site histidine is mutated into any one of lysine, serine and leucine, or / and the 170th-site histidine is mutated into any one of lysine, cysteine and leucine; the carbonyl reductase mutants, especially single-site mutants Mut-K36D and Mut-H125K and a combined mutant Mut-K36D-H125K, with activity improved to different extents compared with that of a wild type are obtained, have excellent thermal stability and relative enzyme activity, have relatively high activity and stability aiming at various chiral alcohols, and can be used for preparing the carbonyl reductase. The method has the advantages of high catalytic activity, high enzyme expression quantity, easiness in fermentation and the like.

The invention relates to a preparation method of D-panthenol. The preparation method comprises the following steps: (1) carrying out mixed reaction on DL-pantoic acid lactone and D-pantoic acid lactone hydrolase, extracting, and separating a water phase to obtain D-pantoic acid; (2) mixing the D-pantoic acid with a compound enzyme auxiliary agent for reaction to obtain D-pantoic acid lactone; (3) mixing the D-pantoic acid lactone with beta aminopropanol for reaction to obtain D-panthenol crude product slurry; (4) purifying the D-panthenol crude product slurry to obtain D-panthenol; the compound enzyme auxiliary agent comprises esterase, carbonyl reductase, alcohol dehydrogenase and alcohol nitrile enzyme. According to the method disclosed by the invention, the esterase, the carbonyl reductase, the alcohol dehydrogenase and the alcohol nitrile enzyme act synergistically to catalyze an esterification reaction in D-pantoic acid molecules and promote the D-pantoic acid molecules to form a lactone structure, so that side reactions are reduced, the efficiency of the esterification reaction of the D-pantoic acid is improved, and finally, the yield and the purity of the D-panthenol are improved.

The invention relates to the technical field of medical intermediates, in particular to a biosynthesis method of carbonyl reductase and a remegapam intermediate. The amino acid sequence of the carbonyl reductase is shown as SEQ ID NO: 1, the carbonyl reductase can be used for synthesizing a remegapam intermediate, and the specific method is as follows: a compound II is taken as a substrate, and in the presence of the carbonyl reductase, a coenzyme, an enzyme circulating hydrogen donor, a coenzyme circulating enzyme, a cosolvent and a buffer solution, a biological catalytic reaction is performed to generate the remegapam intermediate, namely a compound I; the synthesis method disclosed by the invention is mild in reaction condition, simple and convenient to operate, environment-friendly, low in cost, few in by-products and high in ee value of the product, and efficient synthesis of the remegapam intermediate is realized.

The invention relates to a carbonyl reductase mutant based on cosubstrate catalytic ability optimization and application thereof, the mutant is constructed on the basis of a carbonyl reductase BaSDRX amino acid sequence as shown in SEQ ID No.2, the nucleotide sequence of the carbonyl reductase is as shown in SEQ ID No.1 in a sequence table, and the nucleotide sequence of the carbonyl reductase is as shown in SEQ ID No.2 in the sequence table. The mutant is single-point mutation or multi-point combined mutation on the basis of an amino acid sequence of carbonyl reductase BaSDRX as shown in SEQ ID No.2: threonine Thr133 at the 133 site, arginine Arg137 at the 137 site and glutamic acid Glu142 at the 142 site; when the mutant enzyme or a recombinant cell containing the mutant enzyme is applied to synthesis of (S)-1-(2, 6-difluorophenyl) ethanol through asymmetric reduction of 2, 6-difluoroacetophenone, high catalytic activity and high stereoselectivity are achieved, and high-optical-purity (S)-1-(2, 6-difluorophenyl) ethanol (eet; therefore, the method has a good industrial application and development prospect.

The invention relates to the technical field of biology, in particular to a carbonyl reductase mutant, a gene, an engineering bacterium and application. The amino acid sequence of the carbonyl reductase mutant is shown as SEQ ID NO: 1, the catalytic efficiency of asymmetric reductive hydrogenation of prochiral ketone compounds is improved, the stereoselectivity of chiral hydroxyl compounds formed through catalysis is improved, and the synthesis efficiency of (S)-1-(5-fluoro-2-iodophenyl) ethanol is improved.

The invention discloses a carbonyl reductase mutant for synthesizing a chiral alcohol sulfone compound and application of the carbonyl reductase mutant, a mutant M4A1 of carbonyl reductase ChKRED20 is used as a female parent, serine at the 188th site is mutated into any one of asparagine, glycine or threonine, and the carbonyl reductase mutant for synthesizing the chiral alcohol sulfone compound is obtained. Specifically, a mutant M4A1 of ChKRED20 is taken as a female parent, serine at the 188th site is mutated into asparagine, and a mutant ChKRED20-M4C4 is obtained. The mutant M4A1 of ChKRED20 is used as a female parent, serine at the 188th site is mutated into glycine, and the mutant ChKRED20-M4C3 is obtained. The mutant M4A1 of ChKRED20 is used as a female parent, serine at the 188th site is mutated into threonine, and the mutant ChKRED20-M4C6 is obtained. The three mutants can be used as catalysts to synthesize chiral alcohol sulfone compounds, and especially have great advantages in asymmetric synthesis of alpha-substituted beta-alcohol sulfone.

The invention discloses a reductive aminase mutant and application of the reductive aminase mutant in synthesis of chiral alkylated benzyl propylamine. The invention relates to a mutant protein of carbonyl reductase, which is characterized in that the mutant protein has one or more alternative mutations in sites 85, 87, 128, 134, 162, 197, 219, 234 and 235 on the basis of an amino acid sequence as shown in SEQ ID NO: 1. On the basis of analysis of the crystal structure of reductive aminase M5 in the early stage, stereoselective transformation is carried out through site-specific mutagenesis, combinatorial mutation and the like, an efficient mutant for synthesizing an alkylated benzyl propylamine S-enantiomer is obtained, and a series of chiral amines with the selectivity larger than 99% S are synthesized with different amine donors such as benzyl acetone and benzylamine as substrates. And the method has important significance on industrial green synthesis of the chiral amine.

The invention discloses a method for preparing an immobilized enzyme by using nano composite hydrogel and application of the immobilized enzyme, the composite hydrogel is adopted to immobilize the enzyme, negatively charged zymoprotein is enriched on a positively charged nanoparticle carrier through the charge effect, the physical acting force is increased, and a three-dimensional network structure is formed through an in-situ polymerization method; the carbonyl reductase or lipase is immobilized in the hydrogel, so that the pore volume, the specific surface area, the average pore size and the pore width of the immobilized enzyme are improved, the loading capacity of the enzyme is further improved, the maximum loading rate is 3422mg / g, and the problem that the immobilized enzyme is easy to leak by an embedding method is solved. The method is mild in reaction condition, and the stability and catalytic activity of the immobilized enzyme are improved.

The invention provides a method for preparing a (beta, S)-configuration glassine through enzyme catalysis and an engineering bacterium, and belongs to the field of biosynthesis, the engineering bacterium takes Escherichia coli BL21 (DE3) as an original strain, and an improved carbonyl reductase coding gene shown as SEQ ID No.1 and an improved formate dehydrogenase coding gene shown as SEQ ID No.2 are introduced. The enzyme produced by the strain is used for preparing carbonyl reductase LsADH and formate dehydrogenase PsFDHmut co-immobilized enzyme to carry out enzyme catalytic synthesis of (beta, S) configuration bose, the catalytic reaction is high in selectivity and can be carried out in a normal-temperature and normal-pressure neutral environment, the co-immobilized enzyme can be repeatedly used, and continuous production and cost reduction are facilitated. The conversion rate of the adopted catalytic reaction is greater than 99%, the used substrates beta-acetone xyloside and sodium formate are high in safety, and the synthesis efficiency of the product (beta, S) bose is remarkably improved.

The application discloses a carbonyl reductase mutant and application thereof in preparation of (R)-8-chloro-6-hydroxyoctanoate compounds. The application relates to a mutant of carbonyl reductase with improved catalytic performance and stereoselectivity after rational molecular modification, a recombinant expression plasmid containing the mutant gene of the enzyme, a recombinant expression transformant, a preparation method of the recombinant enzyme, and application of the recombinant enzyme or the recombinant cell as a catalyst in asymmetric reduction of 8-chloro-6-carbonyloctanoate compounds (I) to prepare (R)-8-chloro-6-hydroxyoctanoate compounds (II), in particular, in an intermediate (R)-8-chloro-6-hydroxyoctanoate methyl ester of thioctic acid. The disclosed carbonyl reductase SsCR mutant has the advantages of high reaction efficiency and high catalytic activity, and exhibits the advantages of high substrate concentration, low catalyst consumption and short reaction time in the process of catalytic preparation of the compound (II).

The present invention discloses a carbonyl reductase mutant and its application. The mutation sites of the carbonyl reductase mutant of the present invention include the 88th, 142nd, 190th and 193rd positions of the amino acid sequence shown in SEQ ID NO:1. The carbonyl reductase mutant of the present invention has higher enzyme activity than the wild-type carbonyl reductase. The enzyme activity of some carbonyl reductase mutants is 50 times that of the wild-type carbonyl. The carbonyl reductase mutant of the present invention can make the compound shown in formula I undergo a reduction reaction as shown in the following formula in the presence of a coenzyme in a liquid reaction system, and prepare the compound shown in formula ⅠⅠ with a conversion rate of >99%, a chiral ee value of >99%, and a chiral de value of >99%. #imgabs0#

The invention discloses a carbonyl reductase mutant, an immobilized carbonyl reductase and its application, and relates to the field of genetic engineering and enzyme engineering technology. A carbonyl reductase mutant, whose amino acid sequence is shown in SEQ ID NO.1; a carbonyl reductase mutant crude enzyme solution is mixed with chitosan microspheres, a glutaraldehyde solution is added dropwise, oscillation cross-linked, and washed with water to obtain an immobilized carbonyl reductase. The enzyme activity of the immobilized carbonyl reductase prepared in the present application is increased by 5-10 times compared with that of the wild type. When eslicarbazepine is prepared by asymmetric reduction catalyzed by immobilized carbonyl reductase, the substrate concentration reaches 80-150 g / L, the chiral purity value is greater than 99%, and the conversion rate is greater than 99%. The immobilized enzyme of the present invention can be recovered by simple filtration, can be repeatedly used more than 5 times after recovery, and the chiral purity and conversion rate of the reaction used for the first time are substantially the same when reused, and has a high potential for industrial application.

The invention relates to the technical field of biological pharmacy, in particular to a carbonyl reductase mutant and an enzyme catalytic synthesis method of a montelukast sodium intermediate, and the amino acid sequence of the carbonyl reductase mutant is shown as any one of SEQ ID NO: 6-8. According to the enzymatic synthesis method of the montelukast intermediate, a compound II is taken as a substrate, and in the presence of a carbonyl reductase mutant, a coenzyme, a coenzyme recycle hydrogen donor, a cosolvent and a buffer solution, a biological catalytic reaction is carried out to generate the montelukast intermediate, namely a compound I; the specific synthesis route is as follows: # imgabs0 #. The synthesis method is easy and convenient to operate and environmentally friendly, and efficient synthesis of the chiral intermediate is achieved.

The invention relates to an alkene reductase mutant and application of the alkene reductase mutant in synthesis of chiral aromatic polysubstituted gamma-butyrolactone. The alkene reductase mutant is a protein composed of a new amino acid sequence formed by replacing tryptophan at the 100th site of the amino acid sequence as shown in SEQ ID No.2 with alanine. The invention provides a novel alkene reductase mutant SsER-A314Y / W100A, the alkene reductase mutant SsER-A314Y / W100A and a mutant SsCR-V3 (V126A / N210H / M215G) of carbonyl reductase SsCR construct a double-enzyme cascade catalytic system, and the double-enzyme cascade catalytic system is used for efficient stereoselective synthesis of various aromatic chiral gamma-butyrolactone. Compared with the prior art, the aromatic chiral gamma-butyrolactone double-enzyme cascade system provided by the invention has the remarkable advantages of simple process, high atom economy, high optical purity of the product and environmental friendliness, and has a very good industrial application prospect.

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 present invention discloses a carbonyl reductase mutant and a method for catalyzing the synthesis of high-concentration bosogenin, belonging to the field of biocatalysis. The present invention adopts a one-pot method to catalyze β-acetone xyloside to synthesize high-concentration bosogenin, uses an enzyme encoded by carbonyl reductase CnMDR, rationally designs multiple key binding sites of the substrate pocket, combines beneficial mutations, uses a recombinant bacterium with an advantageous mutant enzyme as a catalyst, uses β-acetone xyloside as a substrate, and simultaneously introduces formate dehydrogenase variants and NAD + Kinase, drives NADP + ‑NADPH coenzyme cycle, the yield of synthesized bosone is 200g / L, the conversion rate is>99%, and the product ee value is>99%, which are all the highest levels publicly reported so far. The present invention provides a one-pot method for synthesizing bosone with high efficiency and low cost, which only requires the addition of a trace amount (0.3mM) of coenzyme, and the by-products in the reaction system are easy to separate, which provides great convenience for industrial scale-up 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.