141results about How to "High stereoselectivity" patented technology

The invention belongs to the field of biological pharmacy and biotransformation, and particularly relates to a tagatose-4-epimerase mutant and application thereof. The tagatose-4-epimerase mutant disclosed by the invention is obtained by carrying out single mutation or combined mutation on an amino acid residue at a specific position of an amino acid sequence as shown in SEQ ID NO.1. Compared with wild type tagatose-4-epimerase, the mutant has the advantage that the catalytic activity is improved when the D-tagatose is prepared through conversion. The obtained tagatose-4-epimerase mutant has the advantages of being mild in reaction condition, green, environmentally friendly, single in product and easy to separate and purify, and the production cost is greatly reduced.

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

The invention belongs to the field of biological pharmacy and biological conversion, and particularly relates to a lipase mutant and application thereof. The lipase CALB mutant disclosed by the invention is obtained by carrying out single-point mutation or combined mutation on the 40th site, the 134 site, the 154 site, the 189th site, the 278th site and the 281 site of an amino acid sequence as shown in SEQ ID NO.1. Compared with a wild type, the mutant has the advantage that the stereoselectivity is greatly improved when the mutant is used for catalyzing racemization 3-cyclohexene-1-methyl formate to produce (S)-3-cyclohexene-1-formic acid. Compared with a traditional chemical method for preparing (S)-3-cyclohexene-1-formic acid, the lipase CALB mutant provided by the invention is high in stereoselectivity, mild in reaction condition, environment-friendly, low in equipment requirement, capable of greatly reducing the production cost and wide in application prospect in industrial application.

The invention provides an L-threonine aldolase mutant and application of the L-threonine aldolase mutant in catalytic synthesis of p-methylsulfonylphenylserine, the L-threonine aldolase mutant is derived from pseudomonas, threonine and p-methylsulfonylbenzaldehyde can be used as substrates, and the L-threonine aldolase mutant can be synthesized by using a one-step method. According to the present invention, L-threonine aldolase is adopted as a raw material, pyridoxal phosphate and reduced nicotinamide adenine dinucleotide are adopted as coenzymes, the synthesis of florfenicol intermediate product p-methylsulfonyl phenyl serine is obtained through catalysis, and compared with the wild type L-threonine aldolase, the mutant enzyme has high activity, can effectively improve the conversion efficiency, and can shorten the conversion time; the enzyme and the technical method provided by the invention have the advantages of mild reaction conditions and less pollution, and have good industrial prospects in the production of florfenicol synthesis intermediates.

The invention relates to hydantoinase and application thereof in hydrolysis or preparation of chiral monomeric compounds. Compared with an amino acid sequence as shown in SEQ ID NO: 1, the amino acid sequence of the hydantoin enzyme is different in one or more positions selected from the 63 site, the 64 site, the 70 site, the 92 site, the 94 site, the 123 site, the 318 site, the 319 site and the 345 site. The invention also relates to a nucleic acid molecule, a nucleic acid construct, a recombinant vector and a host cell encoding the hydantoin enzyme, and an enzyme preparation containing the hydantoin enzyme. The invention also relates to a method for hydrolyzing or preparing a chiral monomeric compound by using the hydantoinase. The hydantoinase provided by the invention has better selectivity on substrate configuration and higher hydrolase activity, can catalyze various racemate hydantoin analogues, and selectively hydrolyzes R configuration with high reaction efficiency to obtain a compound chiral monomer with S configuration.

The invention discloses cyclohexene formate esterase, a mutant of cyclohexene formate esterase and application of cyclohexene formate esterase in production of (S)-3-cyclohexene-1-formic acid. According to the cyclohexene formate esterase and the mutant disclosed by the invention, 100-400g / L of racemic 3-cyclohexene-1-methyl formate can be split by taking wet thalli subjected to induced expression or crude enzyme liquid obtained by ultrasonically crushing the wet thalli as a catalyst, so that (S)-3-cyclohexene-1-formic acid with chemical purity and optical purity reaching 99% or above is synthesized. The enzyme activity of the cyclohexene formate esterase mutant provided by the invention is further improved, the reaction time and the catalyst dosage are obviously reduced compared with those of a substrate with the same catalysis concentration before mutation, the reaction condition is mild, the operation is relatively simple and convenient, and the cyclohexene formate esterase mutant has a very good industrial application prospect.

The invention relates to a chiral pyridinium salt carbonyl catalyst as well as a preparation method and application thereof, the structural general formula of the catalyst is # imgabs0 #, in the formula, R1 is # imgabs1 #, X1-6 comprises N or C-R6-11; r2, R4 and R6-11 comprise hydrogen, alkyl of C1-24, alkyloxy, # imgabs2 # CO2Rd, halogen, nitryl, cyano or trifluoromethyl; r3 comprises substituted or unsubstituted C1-24 alkyl groups, substituent groups comprise halogen, C1-10 alkyl groups, C3-10 cycloalkyl or aryl groups, C1-8 carbonyl groups, C1-8 sulfonyl or phosphoryl groups, and C1-10 alkoxy or amido groups, and carbonyl groups comprise aldehyde groups, ketone carbonyl groups, ester carbonyl groups, carboxyl groups or amido groups; and R5 has the structural formula of # imgabs3 # X <-> which is an anion. Compared with the prior art, the catalyst disclosed by the invention is used for a bionic aldol reaction of amino acid derivatives to synthesize a series of beta-hydroxyl-alpha-amino acid compounds with extremely high enantioselectivity.

According to the method for synthesizing the 2-amino-3-hydroxy-4-oxobutyric acid derivative through enzyme catalysis, threonine aldolase LTA and / or threonine transaldolase LTTA are / is adopted for catalysis, and the 2-amino-3-hydroxy-4-oxobutyric acid derivative with double chiral centers can be efficiently and selectively synthesized.

The invention discloses a method for preparing high-purity estra-1, 3, 5 (10)-triene-3, 15 alpha, 16 alpha, 17 beta-tetrol (abbreviated as estratrol), which is characterized in that 3-hydroxy-17 beta-hydroxyestra-1, 3, 5 (10), 15-tetraene is used as a raw material, and the estratrol is prepared by three steps of phenolic hydroxyl group protection, asymmetric dihydroxylation and hydrolysis. According to the preparation method, 3-hydroxy-17beta-hydroxyestra-1, 3, 5 (10), 15-tetraene is taken as a raw material, 3-hydroxyl is protected by alkoxy-substituted benzoyl, asymmetric dihydroxyl reaction is carried out through catalytic oxidation of potassium osmate, double alpha-hydroxyl is introduced to 15, 16-positions, chemical reagents and catalysts with high cost are not needed in the preparation method, and the preparation method is simple and easy to implement. The dihydroxylation has the advantages of high stereoselectivity, easiness in purification and separation, mild reaction conditions and simplicity and convenience in operation, and high-efficiency preparation of high-purity estetrol (more than or equal to 99.5%) can be realized.

The invention discloses a dynamic polyene amine compound as well as a preparation method and application thereof. The structural formula of the dynamic polyene amine compound is as shown in formula (I). The dynamic polyenamine compound is obtained by carrying out polymerization reaction on a binary amide activated alkynyl compound and a diamine compound in an organic solvent. The preparation method has the advantages of mild reaction conditions, no need of catalyst, simple and easily available raw materials, high atom utilization rate, high polymer yield, high molecular weight and excellent stereoselectivity. The dynamic polyene amine compound prepared by the invention has the properties of good processability, film-forming property, high thermal stability, non-traditional luminescence and the like, and the dynamic enamine bond contained in the dynamic polyene amine compound prepared by the invention can realize accurate degradation and structural replacement of a polymer under mild conditions.

The invention provides an R-type transaminase mutant and application thereof, and compared with an amino acid sequence of wild-type transaminase as shown in SEQ ID NO.1, the mutant has mutation of a 62nd site, a 152nd site and a 155th site. The mutant has high catalytic activity and excellent stereoselectivity on a heterocyclic ketone substrate, and can be used for efficiently preparing R-configuration heterocyclic chiral amine.

The present invention discloses a transaminase mutant SEQ ID NO: 3, which can efficiently convert (2Z)-4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7-(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-one into sitagliptin.

The application belongs to the field of biopharmacy and bioconversion, and particularly relates to a lipase mutant and application thereof. The lipase CALB mutant of the application is obtained by adopting single-point mutation or combined mutation on positions 40, 134, 154, 189, 278 and 281 of the amino acid sequence shown in SEQ ID NO. 1. The mutant greatly improves the stereoselectivity in catalyzing the production of (S)-3-cyclohexene-1-methyl formate from racemic 3-cyclohexene-1-methyl formate. Compared with the traditional chemical method for preparing (S)-3-cyclohexene-1-methyl formate, the lipase CALB mutant provided by the application has high stereoselectivity, mild reaction conditions, is friendly to the environment, has reduced requirements on equipment, greatly reduces the production cost, and shows a wide application prospect in industrial application.

The invention provides a method for preparing chiral 3-acetoxymethyl-2,2-dimethylcyclopropanecarboxylate, and belongs to the field of organic synthesis. The invention relates to a method for synthesizing chiral 3-acetoxymethyl-2,2-dimethylcyclopropanecarboxylate from prenyl acetate and diazoacetate through an asymmetric cyclopropanation reaction. The chiral copper catalyst used is generated in situ by copper salt and chiral tridentate P,N,N-ligand in various polar and non-polar solvents. The invention can conveniently synthesize chiral 3-acetoxymethyl-2,2-dimethylcyclopropanecarboxylate, and has the characteristics of simple operation, easy availability of raw materials, wide application range of substrates, high enantioselectivity, etc.

The application relates to the technical field of biotechnology, in particular to a dopamine reuptake inhibitor intermediate synthesis enzyme mutant and application, and amino acid sequences are shown in SEQ ID NO: 7-SEQ ID NO: 12. The stereoselectivity of asymmetric reduction catalysis of the dopamine reuptake inhibitor intermediate precursor is improved, the R-type dopamine reuptake inhibitor intermediate with high chirality purity is obtained, and the synthesis efficiency of the dopamine reuptake inhibitor intermediate is improved.

The invention discloses a planar chiral [2.2] paracyclophane alkane monophosphine ligand pEPCP as well as a preparation method and application thereof, and relates to the technical field of chemical catalysis, the planar chiral [2.2] paracyclophane alkane monophosphine ligand pEPCP is characterized by comprising a racemate or an enantiomer, and the structural formula I of the monophosphine ligand pEPCP is as follows: # imgabs0 #. Different types of ether substituents are introduced into the structure of a unique large-steric-hindrance electron-rich rigid skeleton formed by connecting two benzene rings at para-positions through two ethylidene groups, so that the electron cloud density and steric hindrance of aromatic rings directly connected with phosphine ligands are changed. In addition, the ether substituent can also be used as a weak coordination group and participates in adjusting a dihedral angle of ligand chelation. The metal, the ligand and the substrate form the most appropriate coordination environment in the catalysis process, so that the corresponding catalytic reaction has the advantages of high reaction activity, good enantioselectivity, wide substrate universal application range and the like, is particularly suitable for asymmetric coupling reaction catalyzed by transition metal, and has economic practicability and industrial application prospects.

The invention relates to a pharmaceutical synthesis process of Tovorafenib (Tovorafenib). According to the synthesis process and the related preparation method of the chiral alpha-miscellaneous amine, prochiral ketone is directly reduced and aminated into optical pure chiral alpha-miscellaneous amine by a one-step method under a mild condition by utilizing a biocatalyst amine dehydrogenase, the defects of the existing catalysis technology can be overcome, the process route is simple, the reaction condition is mild, the yield is high, and the method is suitable for industrial production. The method has green synthesis advantages of step economy, atom economy and the like, and has the advantages of good stereoselectivity (ee value is as high as 99%) and the like. In the synthesis route, a method of condensing amino and carboxyl in an HATU and alkaline environment is mostly adopted, the operation is simple, the reaction yield is high, and the method is suitable for batch production.

The present invention discloses a method for preparing an abrocitinib intermediate, and belongs to the technical field of organic synthesis. N 2 The reaction configuration was flipped to prepare cis compound 3. Compound 2 has sufficient stability to avoid S N 1 reaction, solves the technical difficulty of constructing the chiral center, avoids the problem of using rearrangement reaction to generate isomers and urea-type byproducts that are difficult to remove. In addition, compound 3 is deprotected from the p-nitrobenzenesulfonyl protecting group under sodium thioglycolate and potassium carbonate conditions to obtain compound 4; the removal conditions are mild, and there is no need to use strong acid, strong base and heating conditions, avoiding the generation of de-Boc and other byproducts, with good chemical selectivity and high yield. The route of the present invention has high stereoselectivity and chemical selectivity, the product is easy to purify and has a high yield, and the reaction reagents are safe and easy to obtain; the reaction of each step is completed within 12 hours; the total yield and total cost have obvious advantages, and it is suitable for commercial production.

Provided are a lipase mutant and an application thereof. Specifically, one or more mutations selected from A262H, A338V, V364I, A158P / V, and I159N are generated on the basis of an amino acid sequence as shown in SEQ ID NO: 1; and compared with a parental lipase, a change in the structure and function of a protein occurs in the lipase mutant, and the stereoselectivity is improved, such that a usage amount of an enzyme is decreased to a certain extent, the post-treatment difficulty is reduced, and the lipase mutant is suitable for industrial production.

The invention relates to the technical field of biology, in particular to a biological preparation method of a remegapam intermediate, which comprises the following steps: by taking remegapam intermediate precursor ketone with a certain concentration as a substrate, adding recombinant engineering bacteria, reacting in a reaction system with the pH value of 8.0-9.0 at the temperature of 30-40 DEG C, and obtaining a product after the reaction is completed, wherein the recombinant engineering bacterium is an engineering bacterium containing a ketoreductase mutant coding gene, and the amino acid sequence of the ketoreductase mutant is as shown in SEQ ID NO: 4, SEQ ID NO: 19 or SEQ ID NO: 39. According to the biological preparation method of the remegapam intermediate disclosed by the embodiment of the invention, the stereoselectivity of asymmetric reduction catalysis of the used ketoreductase mutant on precursor ketone of the remegapam intermediate is improved, the R-type remegapam intermediate with high chiral purity is obtained, and the synthesis efficiency of the remegapam intermediate is favorably improved.

The application provides a ketoreductase mutant and application thereof in synthesis of a key chiral intermediate of tigogaral, and provides a ketoreductase mutant with an amino acid sequence as shown in SEQ ID NO: 4, 6, 8, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40 or 42. The ketoreductase mutant has higher carbonyl catalytic activity and chiral selectivity than a wild type, and is used for preparation of a key intermediate R-5,7-difluorobenzodihydropyran-4-ol of tigogaral.

The invention provides a method for preparing chiral amine by catalyzing asymmetric hydrogenation of aromatic imine through a manganese / chiral ferrocene P, N, N ligand. The manganese precursor and ligand adopted by the method are easy to obtain, the reaction condition is mild, the operation is simple, the target chiral amine compound can be obtained with high yield and high stereoselectivity, and continuous large-scale preparation can be realized. The catalyst system is high in activity, the molar ratio of imine to the catalyst can reach 100000: 1, and the catalyst has a very good industrial application prospect.

This invention provides an imine reductase mutant, its preparation method, and its application in the catalytic preparation of dextromethorphan intermediates. The imine reductase mutant is an imine reductase with an amino acid mutation, comprising the amino acid sequence shown in SEQ ID NO:1. The amino acid mutation type includes any one or a combination of at least two of I122T, D212A, or G228R. By introducing a mutation into the imine reductase, this invention significantly improves the enzyme's activity and stereoselectivity, enabling the high-yield synthesis of the dextromethorphan intermediate (S)-1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline under mild conditions, greatly reducing production costs and making it suitable for industrial production.

This invention discloses a chiral monophosphine ligand, its preparation method, and its application in asymmetric synthesis. The ligand preparation method involves mixing an N,N',1,2-tetrasubstituted ethane-1,2-diamine compound, phosphorus trichloride, triethylamine, and solvent under a nitrogen or argon atmosphere at -80 to -70°C, and reacting at 60 to 80°C for 6 to 8 hours. The resulting reaction mixture is then cooled to room temperature, and an alcohol, phenol, or Grignard reagent is added dropwise, reacting at 60 to 80°C for 8 to 12 hours. The resulting mixture is then post-treated to obtain the chiral monophosphine ligand. The application involves stirring a nickel catalyst, a chiral monophosphine ligand, a solvent, a diene, and an aldehyde under an argon or nitrogen atmosphere at a specified temperature for 12 to 15 hours. The mixture is then post-treated to obtain the target product. This method utilizes widely available and inexpensive raw materials, and the operation is simple, achieving high stereoselectivity and enantioselectivity in the synthesis of alcohol compounds containing chiral conjugated dienes through a one-step reaction.

The invention provides a dianhydroidurol-based diamino compound and a preparation method thereof, and the preparation method comprises the following steps: obtaining a dianhydroidurol-containing dinitro compound based on dianhydromannitol or dianhydrosorbitol, dissolving the dianhydroidurol-containing dinitro compound in ethyl acetate, and reacting to obtain the dianhydroidurol-based diamino compound. And carrying out a reduction reaction in a hydrogen atmosphere to obtain the diamino compound based on the dianhydroidurol. The invention further provides a PI film prepared from the diamino compound based on the dianhydroidurol and a preparation method of the PI film, and the PI film is obtained by doping the diamino compound based on the dianhydroidurol with 3, 4 '-ODA and then adding dianhydride for polycondensation. By introducing the dianhydroidurol, destroying the conjugated structure of the chain segment, weakening the interaction between molecular chains and increasing the distance and free volume between the molecular chains, the polarizability of molecules is reduced, and finally the purposes of enhancing the light transmission and reducing the dielectric constant are achieved.

The invention relates to the technical field of organic synthesis, in particular to a synthesis method of N-Boc protected L-beta-homopiperidine acid methyl ester. According to the invention, cheap and easily available L-2-amino adipic acid is used as an initial raw material, and the target product N-Boc protected L-beta-homopiperidine acid methyl ester is finally prepared through conversion of five steps. The synthetic route is convenient to operate, good in stereoselectivity, mild in reaction condition, simple in separation and purification, relatively high in total yield and capable of realizing large-scale preparation. The adopted raw materials and reagents are non-toxic, the production process is pollution-free and environment-friendly, and good conditions are created for industrial large-scale production and commercialization of the product.

This invention discloses a mutant amine dehydrogenase and a method for synthesizing non-α-amino acids, belonging to the field of biocatalysis engineering. This invention selects keto esters as substrates and utilizes phenylalanine amine dehydrogenase to complete the asymmetric reductive amination of these esters with different substitution positions. A strategy combining substrate engineering and enzyme evolution is employed to enhance substrate-enzyme compatibility, increasing the activity of β-, γ-, δ-, and ε-keto esters by 13-fold, 8-fold, 9-fold, and 40-fold, respectively. Results show that substrate engineering and enzyme evolution promote nonpolar interactions between the enzyme and substrate, improving substrate binding affinity. Using superior mutants, various non-α-amino acids were synthesized with high conversion rates and enantioselectivity exceeding 99%. The synthesized amino acids include β- and γ-amino acids, as well as δ- and ε-amino acids with more distant substitution positions. Therefore, an eco-friendly enzyme chemistry cascade has been successfully established for the synthesis of non-α-amino acids with different substitution patterns.

The invention discloses a method for preparing a chiral geminal diaryl methylene silane compound, using N-aryl sulfonyl hydrazone compounds, aryl bromides and (dimethylphenylsilyl) boric acid pinacol ester as raw materials, and enantioselective three-component coupling is carried out under the co-catalysis of palladium and copper to prepare chiral geminal diaryl methylene silane. The above preparation method has mild reaction conditions, easy to obtain reactants and good functional group tolerance, and the synthetic product has high stereoselectivity and high yield, and can quickly and efficiently prepare the enantiomer of the target compound. In addition, the method has a wide range of substrates, can be used to prepare chiral silanes that cannot be obtained by carbene Si-H insertion method, and provides new ideas for designing and synthesizing novel chiral organosilane compounds.

The invention discloses a carbonyl reductase mutant and an application of the carbonyl reductase mutant in preparation of a (2R, 3R)-2-fluoro-3-hydroxyl ester compound. In particular to a preparation method for preparing the (2R, 3R)-2-fluoro-3-hydroxy ester compound by using carbonyl reductase derived from Zygosaccharomyces bayer and free enzyme or recombinant expression transformant of a mutant of which the enzyme activity and the stability are remarkably improved as a catalyst through asymmetric reduction by using the carbonyl reductase derived from Zygosaccharomyces bayer and the free enzyme or the recombinant expression transformant of the mutant as the catalyst. Compared with other methods for preparing (2R, 3R)-2-fluoro-3-hydroxy ester compounds, the enzyme and the technical method thereof disclosed by the invention have the advantages that the theoretical yield is up to 100%, the reaction condition is mild, the enzyme is environment-friendly, the operation is simple and convenient, the industrial amplification is easy, the substrate concentration is high, and the like, and the enzyme and the technical method thereof can be used for preparing (2R, 3R)-2-fluoro-3-hydroxy ester compounds. The compound has a good application prospect in the (3R, 3R)-2-fluoro-3-hydroxy ester compound.

The invention relates to the technical field of catalytic organic synthesis, in particular to calix [4] arene cyclohexanediamine derivatives and a method for green catalysis of asymmetric Aldol reaction by the calix [4] arene cyclohexanediamine derivatives. The method comprises the following steps: by taking aromatic aldehyde and cyclic ketone as raw materials, calix [4] arene cyclohexanediamine derivative as a phase transfer catalyst, p-nitrobenzoic acid as an additive and water as a solvent, reacting at 25 DEG C for 24-48 hours, adding dichloromethane after the reaction is finished, extracting and separating to obtain an organic phase, drying by using anhydrous Na2SO4, and separating and purifying by column chromatography to obtain an Aldol product. The calix [4] arene cyclohexanediamine derivative is mild in synthesis process condition and high in catalytic reaction efficiency, asymmetric Aldol reaction catalyzed by the calix [4] arene cyclohexanediamine derivative takes water as a solvent, the calix [4] arene cyclohexanediamine derivative is green and environment-friendly, good dr value and ee value can be obtained through catalytic reaction at room temperature, and the calix [4] arene cyclohexanediamine derivative has a wide application prospect.