126 results about "Chiral amine" patented technology

The invention discloses an omega-transaminase progenitor enzyme mutant for efficient catalytic synthesis of (R)-chiral amine compounds and application, and relates to the technical field of molecular biology. The omega-transaminase mutant is obtained by mutating an ancestor enzyme Ancaa-101 of omega-transaminase from Aspergillus terreus, and the amino acid sequence of the omega-transaminase mutant is as shown in SEQ ID NO. 6. The invention also discloses a preparation method of the omega-transaminase mutant. Compared with an ancestor enzyme, the activity of the omega-transaminase ancestor enzyme mutant on a non-natural substrate, namely 1-acetyl naphthalene, N-tert-butyloxycarboryl-3-piperidone or 3-oxopyrrolidine-1-carboxylic acid tert-butyl ester, is remarkably improved.

The invention discloses a chiral amine reagent and a preparation method thereof as well as a method for analyzing and splitting a chiral drug, the chiral amine reagent is APMA # imgabs0 # or d5-APMA # imgabs1 #, and the configuration of a carbon atom connected with an amino group on a piperidine ring in the structural formula is an S configuration. The APMA and d5-APMA provided by the invention can be used for analysis and detection of chiral drugs, 3-aminocyclohexyl-phenyl methane ketone (APMA) and the d5-APMA in a stable isotope form thereof are independently or jointly used as a pair of isotope labeling reagents and are subjected to condensation reaction with the chiral drugs to obtain derivatives, enantiomers of the chiral drugs can be effectively split, and the chiral drugs can be analyzed and detected. And an easy-to-ionize group is introduced, so that the mass spectrum response in an ESI positive ion mode is greatly improved.

The present application relates to imine reductases useful in the synthesis of high optical purity amines, and specifically provides engineered polypeptides having improved imine reductase activity and / or stereoselectivity compared to wild-type sequences, and further, to the synthesis of high optical purity amines. The invention provides a fusion protein of the engineered polypeptide, a polynucleotide, a nucleic acid construct, an expression vector, a host cell, a composition and a method for catalytically synthesizing chiral amine by using the engineered polypeptide.

The invention belongs to the technical field of biological catalysis, and particularly relates to application of R-transaminase and a mutant thereof in asymmetric synthesis of chiral amine compounds. The amino acid sequence of the R-transaminase is as shown in SEQ ID NO. 1. The amino acid sequences of the R-transaminase mutants are respectively shown as SEQ ID NO: 2 (G66L), 3 (F127A), 4 (F127M), 5 (G66L / F127M), 6 (G66L / F127A), 7 (G66L / F127M / L160I) and 8 (G66L / H67N / F127M / L160I). The recombinant transaminase mutant obtained by the invention has the advantages of high catalytic activity, strict stereoselectivity, mild reaction conditions and the like for various cyclic ketone and N-heterocyclic ketone compounds.

The invention provides a transaminase mutant and application of the transaminase mutant in chiral amine synthesis. According to the transaminase derived from Mycobacterium sp. Provided by the invention, in a 1 mL reaction system for catalyzing N-Boc-3-piperidone to generate N-Boc-3-aminopiperidine, the 24-hour conversion rate is 95%, and the e.e. Value of the product is greater than 99%. On the basis, the invention further provides a series of mutants of the transaminase, the relative enzyme activity of the mutants reaches 153%-293%, and the catalytic activity of the transaminase is further improved. Wherein the 34-hour conversion rate of the optimal transaminase mutant is increased to 90% or above under the condition of 120g / L substrate concentration in a 10mL reaction system, and the e.e. Value of the product is greater than 99%. The production technology has the advantages of high catalytic activity, easiness in fermentation, environment friendliness and the like, and has good development and application values.

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.

A method for recovering 4-BMA from a 4-BMA mother liquor reclaimed material belongs to the field of synthesis of medical intermediates, and comprises the following steps: 1) adding the 4-BMA mother liquor reclaimed material into a solvent, heating and dissolving; 2) adding a chiral amine reagent to precipitate 4-BMA in the recycled material, and filtering to obtain 4-BMA salt; 3) adding alkali into the 4-BMA salt for dissociation, and extracting the dissociated chiral amine reagent; and 4) adding acid to separate out 4-BMA, filtering and drying to obtain qualified 4-BMA. According to the method, the chiral amine reagent and 4-BMA are used for selective salification and crystallization, and isomer impurities are not salified and are continuously dissolved in the solvent, so that the product is purified, and the method has the characteristics of high recovery rate, good product quality and simple method, is obvious in reliability and economical efficiency, has obvious cost advantage and is suitable for industrial production.

The invention relates to an organic crystalline piezoelectric material based on chiral 4-bromophenylethylamine hydrochloride, which is synthesized by using hydrochloric acid as an anion and protonated (R / S)-4-bromophenylethylamine as a cation according to the stoichiometric ratio of (R / S)-4-bromophenylethylamine to hydrochloric acid of 1: 1. The invention provides an organic crystalline piezoelectric material based on chiral 4-bromophenylethylamine hydrochloride and a preparation method, and provides a two-dimensional hydrogen bond network construction strategy based on protonated chiral amine: protonated (R / S)-4-bromophenylethylamine cations are used as structural elements, chloride ions are used as charge balance bodies, and the chiral 4-bromophenylethylamine hydrochloride is prepared by precisely regulating the interaction of intermolecular hydrogen bonds. Constructing a two-dimensional hydrogen bond network with high orientation; by enhancing ordered arrangement of molecular dipoles and optimizing a lattice strain-polarization response collaborative mode, breakthrough improvement of piezoelectric performance is successfully realized, and a new normal form is provided for developing high-performance organic piezoelectric materials.

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 present invention discloses a transaminase and its use in preparing optically pure chiral amines. The amino acid sequence of the transaminase of the present invention has one or more mutations at positions 65, 318, 399, 419, 426, 465, and 466 compared to SEQ ID NO: 1, wherein the mutation is an addition, deletion, or substitution of an amino acid residue. The present invention also discloses a nucleic acid molecule encoding the transaminase, a nucleic acid construct comprising the nucleic acid molecule, a recombinant vector, and a host cell. The present invention also discloses an enzyme preparation comprising the transaminase of the present invention. The present invention also discloses a method for preparing chiral amines using the transaminase. In the industrial production process of optically pure chiral amines, the enzyme provided by the present invention has advantages such as substrate specificity, enantiomeric selectivity, and high conversion rate. The method for preparing optically pure chiral amines provided by the present invention has high reaction efficiency, stereoselectivity, and yield.

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.

Provided are a transaminase mutant and use thereof. The transaminase mutant is obtained by mutation occurred in an amino acid sequence as shown in SEQ ID NO:1, and the mutation includes at least T291A; or an amino acid sequence of the transaminase mutant has the mutation in the mutated amino acid sequence, and has more than 80% of identity with the mutated amino acid sequence, preferably has 85% of identity, and more preferably has 90% of identity. The transaminase mutant expands a substrate spectrum of enzymes, improves stereoselectivity, and improves the catalytic capacity of a large steric hindrance substrate; and at the same time, the stability of the enzymes is improved, and a catalytic reaction may be performed to produce chiral amines at high solvent concentrations and high temperatures.

The embodiment of the invention discloses an imine reductase mutant as well as a coding gene and application thereof, and belongs to the technical field of genetic engineering and biosynthesis, and the similarity between the amino acid sequence of the imine reductase mutant and the amino acid sequence of wild type imine reductase is 90%-99.9%. Wherein the amino acid sequence of the wild type imine reductase is as shown in SEQ ID NO. 1. According to the imine reductase mutant provided by the invention, the catalytic activity, optical selectivity and stability of the imine reductase mutant during synthesis of chiral amine compounds can be improved.

A Pseudomonas putida ω-aminotransferase gene with broad substrate spectrum and an application thereof are provided. According to the disclosure, codon optimization is carried out on an ω-transaminase gene, and the nucleotide sequence of the ω-transaminase gene is shown in SEQ ID NO: 1. The crude enzyme solution, crude enzyme powder and pure enzyme of ω-aminotransferase prepared from ω-aminotransferase gene can be used as an enzyme preparation to catalyze the transamination reaction of substrates to prepare chiral amino alcohols and chiral amines. The substrate spectrum of the recombinant ω-transaminase obtained by the disclosure is broad, and 15 kinds of chiral amino alcohols and chiral amines with high added value are asymmetrically catalyzed and synthesized; in particular, 4-hydroxy-2-butanone is catalyzed to produce pharmaceutical intermediate (R)-3-amino-1-butanol.

The present disclosure provides engineered transaminase polypeptides useful for the synthesis of chiral amine compounds under industrially relevant conditions. The disclosure also provides polynucleotides encoding the engineered transaminase polypeptides, host cells capable of expressing the engineered transaminases, and methods of using the engineered transaminases for the production of chiral amine compounds.

The present invention provides a method for synthesizing chiral amines. The method for synthesizing chiral amines provided by the present invention comprises: subjecting a carbonyl substrate and an amine donor to a reductive amination reaction under the catalytic action of a catalyst to obtain a chiral amine, wherein the catalyst is an imine reductase or a reductive aminase. In the present invention, imine reductase or a reductive aminase is used as a catalyst for the synthesis of chiral amines, and a non-keto acid ketone compound substrate and an amino donor can be synthesized into a chiral amine under mild reaction conditions, which not only has a wide substrate range, but also is simple and economical to operate, and can directly synthesize a variety of chiral amine compounds, and can be suitable for industrial production.

The present application provides a method for preparing a captopril intermediate D-3-acetylmercapto-2-methylpropionic acid, and relates to the pharmaceutical field. The method for preparing the captopril intermediate D-3-acetylmercapto-2-methylpropionic acid comprises: reacting a chiral amine with racemic 3-acetylmercapto-2-methylpropionic acid to obtain a chiral amine salt, wherein the chiral amine salt is subjected to alkaline hydrolysis to obtain sodium D-3-acetylmercapto-2-methylpropionic acid, and then acidified to obtain D-3-acetylmercapto-2-methylpropionic acid; wherein the chiral amine comprises one or more of R(+)-alpha-methylbenzylamine, (R)-(+)-1-amphetamine, and R(+)-alpha-tert-butylbenzylamine. The method for preparing the captopril intermediate D-3-acetylmercapto-2-methylpropionic acid provided in the present application comprises splitting 3-acetylmercapto-2-methylpropionic acid with a chiral amine, and the method is controllable and suitable for large-scale production.

The present application relates to a method of preparing ethyl (R)-4-((1-(2-cyanoacetyl)piperidin-3-yl)amino)-1H-pyrrolo[2,3-b]pyridine-5-carboxylate (Compound 1). The process involves the addition of an ethyl carboxylate group, displacement of a halogen with a chiral amine, deprotection of the chiral, cyclic amine, and a subsequent coupling reaction with cyanoacetic acid. The displacement may be conducted in an autoclave at elevated pressure.

Provided are a method for preparing (R)-ketorolac and use thereof. The method includes the following steps: step 1, preparing ketorolac; and step 2, subjecting the ketorolac to resolution to obtain the (R)-ketorolac, the resolution being selected from the group consisting of chiral amine resolution and enzyme resolution.

The invention discloses an amine dehydrogenase catalytic system, which comprises a deep eutectic solvent and a buffer solution, the deep eutectic solvent takes choline chloride or betaine as a hydrogen bond acceptor and carboxylic acid ammonium salt as a hydrogen bond donor; the catalytic system is applied to a reaction for synthesizing corresponding chiral amine from ketone, especially beta-position heteroaryl ketone, so that the conversion rate and the yield are remarkably improved. The eutecticevaporate solvent is non-toxic, harmless, easy to operate and easy to degrade, the enzyme activity and stability of amine dehydrogenase and glucose dehydrogenase in the ketoreductive amination process can be successfully improved, optimization of an amine dehydrogenase catalytic system is achieved, and development and application of a green, convenient and low-cost ketoreductive amination reaction method are facilitated.

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.

The invention discloses an imine reductase mutant and an application of the imine reductase mutant in synthesis of vibeglone. The invention discloses an imine reductase mutant, which is characterized in that on the basis of an imine reductase mutant IR-13-M5, the imine reductase mutant has the following mutations: M203P-F269W-I149G-S241Q-L200T-G268Q, M203P-F269W-I149G-L200T, M203P-F269W-I149G-S241Q-L200T, M203P-F269W-I149G-S241Q-L200T or M203P-F269W-I149G-L200T-G268Q, and the imine reductase mutant has the following mutations: M203P-F269W-I149G The mutant can be used for asymmetric catalytic synthesis of an S-selective chiral amine product vibeglone, the conversion rate reaches 78%, the stereoselectivity reaches up to 99%, and the mutant has a good industrial application prospect.

The invention provides mutant monoamine oxidase and application thereof in synthesis of chiral amine compounds. Specifically, the invention provides monoamine oxidase or a mutant thereof capable of efficiently catalyzing a desymmetric oxidation reaction of 3-azabicyclo [3.3. 0] octane. The monoamine oxidase or the mutant of the monoamine oxidase can be used for efficiently catalyzing 3-azabicyclo [3.3. 0] octane to be subjected to desymmetric oxidation to form (3S, 7R)-3-azabicyclo [3.3. 0] octyl-2-ene, so that the monoamine oxidase or the mutant of the monoamine oxidase can be used for producing chiral amines such as telaprevir intermediates or other chiral amine precursors.

The present invention discloses a rapid synthesis method for axially chiral aromatic amine compounds using metal-free catalysis. 1-substituted tetrahydro-2-naphthalenone and a cheap and readily available chiral amine are used as raw materials. Axial chiral aromatic amines are prepared by reaction under the action of a catalyst, and a wide range of substrates can be selected. Compared with traditional synthesis methods, the method of the present invention uses metal-free catalysis, has the advantages of simple conditions, convenient operation, relatively high yield, low environmental pollution, and ease of industrial production. The synthesis method of the present invention can simultaneously obtain two axially chiral aromatic amine compounds, and the yield range is 20%-70%.

The invention provides an improved engineered phenylalanine ammonia lyase which can be used for preparing a chiral amine compound, in particular for asymmetrically synthesizing L-2-chloro-phenylalanine. Compared with a wild-type enzyme corresponding to SEQ ID NO: 2, the engineered phenylalanine ammonia lyase polypeptide has better activity and / or stability, can be used for asymmetrically preparing chiral amine compounds with extremely high stereoselectivity, particularly can be used for more efficiently catalyzing o-chloro-cinnamic acid to prepare L-2-chloro-phenylalanine, can accept various ammonia donors, and can be used for preparing the chiral amine compounds. Such as ammonium carbonate, ammonia water and ammonium chloride.

The invention discloses a universal covalent organic framework material with spiral chiral morphology and a synthesis method thereof. The covalent organic framework material with spiral chiral morphology is generated by introducing a chiral amine precursor and multilateral aromatic aldehyde molecules to realize imine condensation under an acid catalysis condition. By changing the molecular chirality of the used chiral amine precursor, the regulation and control of the spiral direction of the chiral morphology of the finally obtained covalent organic framework material can be realized. According to the method, the covalent organic framework material with high crystallinity, large specific surface area and uniform spiral morphology can be synthesized under the catalysis of various solvents and various acids. The method has good synthesis universality, and the synthesized covalent organic framework material with the spiral chiral morphology has good chiral optical properties.

This invention provides a method for preparing chiral N-tert-butylsulfonamide compounds. The method uses N-tert-butylsulfonylimide as a starting material, employs Mn(CO)₅Br and a chiral PNN ligand as catalysts, and an alcohol as a solvent. A hydrogenation reaction occurs in this homogeneous catalytic system, yielding axially chiral diaryl nitrophenol compounds in high yield (99%) and with high ee values ​​(>99%). This reaction has the advantages of high efficiency, mild conditions, and environmental friendliness. The obtained chiral N-tert-butylsulfonamide can be deprotected with a sulfonyl protecting group to obtain the corresponding chiral primary amine.

The present invention relates to proteins having improved lipase activity, nucleic acid molecules encoding the corresponding proteins having improved lipase activity, and methods for the stereoselective synthesis of chiral amines or the increase of chiral amine isomers in enantiomer mixtures.

The present invention discloses an imine reductase mutant, a recombinant genetic engineering bacterium and their application in the synthesis of (R)-2-methylpyrrolidine. The imine reductase mutant uses the wild-type imine reductase IR23 with the amino acid sequence shown in SEQ ID NO.1 as a template, and the mutant V249T obtained by mutating valine at position 249, and the mutant V249T / W224F obtained by mutating valine at position 249 and tryptophan at position 224. Through the molecular modification method of semi-rational design, the present invention significantly improves the ability of imine reductase to catalyze the formation of the corresponding cyclic amine from the cyclic imine 2-methylpyrroline. When the mutant V249T / W224F catalyzes 2-methylpyrroline, the substrate conversion rate is 93.3%, which is increased by 49.8% compared with the wild type. The present invention effectively improves the ability of imine reductase to asymmetrically synthesize chiral amines, which is of great significance for the green catalytic synthesis of chiral amine compounds.

The invention discloses a circular polarization organic laser material and a preparation method and application thereof, and the preparation method of the circular polarization organic laser material comprises the following steps: taking chiral amine, rare earth chloride and hydrochloric acid as raw materials, placing the raw materials in a solvent, and heating for reaction to obtain a precursor solution; after drying the precursor solution, dissolving again to obtain a transparent solution; and putting the open container filled with the transparent solution into a closed container filled with an anti-solvent, and growing the zero-dimensional perovskite structure luminescent material A4BX7 single crystal by utilizing the diffusion of the anti-solvent. The chiral ligand is introduced into the hybrid perovskite, a chiral environment is successfully introduced into an inorganic perovskite system, chiral transfer is realized, and the prepared circular polarization organic laser material has good circular polarization organic laser properties and high fluorescence quantum yield.