37 results about "L-2-Aminobutyric Acid" patented technology

The invention discloses a method for producing L-2-aminobutyric acid, and the method comprises the following step: catalyzing L-threonine utilized as a raw material through an enzyme catalysis system consisting of threonine deaminase, L-amino acid dehydrogenase and coenzyme regeneration systems, thus producing the L-2-aminobutyric acid. The method for producing theL-2-aminobutyric acid has the advantages that the raw material is low in price, the property is stable, and the production cost of the L-2-aminobutyric acid can be greatly lowered, the conversion rate and product concentricity are high, no influence caused by byproducts exists, and the method is suitable for industrialization application.

The invention relates to a vector, an engineering strain and a method for producing L(+)-2-aminobutyric acid. The vector is a recombinant vector containing a threonine dehydratase coding gene, an L-amino acid dehydrogenase gene and appropriate vector segments. The engineering strain is obtained by transforming host bacteria by the recombinant vector. The L(+)-2-aminobutyric acid is prepared by fermentation culture of the engineering strain. The engineering strain is fermentation-cultured by glucose as a raw material to produce L(+)-2-aminobutyric acid. The method has the advantages of low cost, high product concentration, no by-product, easy product purification, and good industrial application feasibility.

The invention establishes aminopherase through genetic engineering. Compared with ochrobactrum anthropi based wild w-aminopherase coming from ochrobactrum anthropi, the enzyme activity of aminopherase is remarkably improved, and aminopherase can be used for industrially producing L-2-aminobutyric acid.

The invention discloses a construction and application of a recombinant strain converting L-threonine to L-2-aminobutyric acid, belonging to the field of bioengineering technology. The production method of the invention utilizes a recombinant bacterium expressing two plasmids to simultaneously realize the high-efficient expression of three enzymes, and comprises the steps of: conversion of L-threonine to L-2-aminobutyric acid, coupled with a coenzyme regeneration system, converts NAD+ into NADH, so that the concentration of NADH in the system is relatively stable, and the conversion can be carried out efficiently. Furthermore, the CO<2> converted from ammonium formate can be dissolved in ammonia water, which has little environmental pollution and industrial application value. The method has the advantages of mild conversion condition, strong specificity, low cost and short conversion time. The L-2-aminobutyric acid is prepared adopting the method, 40g/L L-threonine is added, The concentration of the obtained product L-2-aminobutyric acid is 43.3 g/L, and the conversion ratio was over 99.9%.

The invention provides a production method of S-(+)-2-aminobutanamide hydrochloride which is a key intermediate necessary for synthesizing anti-epileptic chiral drug, namely levetiracetam. The method comprises the following steps of: using L-threonine as an initial raw material, preparing L-2-aminobutyric acid by using a biotransformation method, and then performing esterification and ammonolysis reaction to obtain a target compound. According to the method provided by the invention, biotransformation and chemical synthesis are combined, and the good optical selectivity of the biotransformation is utilized for generating the L-2-aminobutyric acid, the reaction condition is mild and simple, the raw material cost is low, an organic solvent is not necessary, the process is environment-friendly, the purity of an isomer is high, and both the conversion rate and the yield are high; and therefore, the S-(+)-2-aminobutanamide hydrochloride is suitable for industrial production.

The invention relates to a method for producing L-2-aminobutyric acid by fermentation, comprising the following steps: (1), inserting the strain into a seed tank to obtain a mature strain; (2), adding the mature strain Connect it into a fermenter for fermentation and cultivation; (3) After the fermentation is over, the fermentation liquid is first sterilized by a ceramic membrane, then passed through an ultrafiltration membrane with a molecular weight of 2-4Kd, and decolorized by activated carbon, and the 732 ion exchange resin is used Adsorbed and eluted with ammonia water, evaporated and concentrated to obtain the crude product of L-2-aminobutyric acid. The method for producing L-2-aminobutyric acid of the present invention, through the improvement of fermentation formula and process control, microbial enzyme conversion scheme and control process, extraction scheme and control process, makes the extraction process simple and the product purity can reach more than 95%, The production capacity and efficiency of L-2-aminobutyric acid are greatly improved, which can meet the large-scale industrial production of L-2-aminobutyric acid, and have higher social and economic benefits.

The invention discloses a method for synthesizing L-2-aminobutyric acid by an enzymatic method. According to the method, 2-ketobutyric acid is catalyzed by the aid of alanine dehydrogenase and formate dehydrogenase to produce the L-2-aminobutyric acid. The method particularly includes the steps: uniformly mixing 0.5-1.5mol of 2-ketobutyric acid, 0.5-1.5mol of ammonium formate, o-0.5g/L of NAD (nicotinamide adenine dinucleotide) and 20g/L of formate dehydrogenase and alanine dehydrogenase co-expression wet cells (or 10-30g/L of formate dehydrogenase wet cells, 10-30g/L of alanine dehydrogenase wet cells) in a rector; adjusting a pH (potential of hydrogen) value to be 7.0-9.0; performing catalytic reaction for 16-20h at the temperature ranging from 30 DEG C to 37 DEG C to obtain the L-2-aminobutyric acid. The method is rapid in reaction, reverse oxidation deamination activity is low, and product loss is greatly reduced.

The invention relates to a method for preparing L-2-aminobutyric acid through whole-cell bioconversion. The method disclosed by the invention comprises the following steps of: carrying out tandem expression on threonine deaminase and leucine dehydrogenase, and cloning on a chlorampenicol resistant expression vector so as to realize co-expression of the two enzymes in the same bacterium; cloning formilase on a kalamycin resistant pET-28a vector to realize expression, and carrying out tandem expression with a pcnB gene in a rate-limiting step of coenzyme I metabolic pathways in escherichia coli so as to realize high expression of the two enzymes; converting the expression vector having two resistances into the same bacterium to carry out co-expression so as to realize expression of four enzymes in the same host bacterium; and carrying out fermentation expression of whole-cell for escherichia coli containing the four enzymes by using escherichia coli so as to realize biological reduction conversion from threonine to L-2-aminobutyric acid. According to the invention, high-efficiency conversion of 2-ketobutyric acid can be realized without addition of coenzyme; the cost is low; and the method is simple.

The invention discloses a threonine deaminase mutant as well as a preparation method and application thereof. The mutant is prepared by carrying out single-point mutation or multi-point mutation on amino acids on a site 14, a site 323, a site 344, a site 449 and a site 510 in an amino acid sequence represented by SEQ ID NO.1. Compared with wild threonine deaminase, threonine deaminase provided bythe invention has the advantages that the stability is extremely high, and the service life of threonine deaminase in the industrial production can be greatly prolonged. A G323D/F510L/T344A mutant isapplied to the production of L-2-aminobutyric acid, the conversion rate of L-threonine can reach 99%, and the addition amount of NAD<+> can be decreased to 0.04g/L, so that a good technical support isprovided for the large-scale production of L-2-aminobutyric acid.

The invention discloses a method for preparing L-2-aminobutyric acid by an asymmetric conversion method. The method comprises the following steps: mixing DL-2-aminobutyric acid with D-tartaric acid; in the presence of aromatic aldehyde as catalyst, reacting in acid solvent; cooling, filtering, and re-crystallizing to obtain a reaction intermediate product; mixing the reaction intermediate product with an aminolysis agent to be subjected to aminolysis reaction; and then cooling, filtering and washing to obtain the L-2-aminobutyric acid. The L-2-aminobutyric acid prepared by the method has the advantages of high yield, higher optical purity and chemical purity, lower cost and suitability for large-batch industrial production.

The invention provides a method for preparing L-2-aminobutyric acid through biocatalysis. The method comprises the following steps: establishing genetically engineered bacteria for simultaneously expressing FDH (Formate Dehydrogenase) and LDH (Leucine Dehydrogenase) to obtain an FDH/LDH co-expression crude enzyme solution containing the FDH and the LDH through fermentation; preparing a reaction system in a buffer solution, and performing biocatalytic reaction to prepare L-2-aminobutyric acid, wherein the reaction system comprises threonine, ammonium formate, a TD enzyme solution, the FDH/LDH co-expression crude enzyme solution, NAD<+> and pyridoxal phosphate. Through the adoption of the method, the preparation of L-2-aminobutyric acid can be simple, efficient and low in cost.

The invention discloses a recombination co-expression system for preparing L-2-aminobutyric acid and application thereof and belongs to the technical field of genetic engineering. The recombination co-expression system is a single-plasmid double-gene expression carrier or a double-plasmid co-expression system, wherein the single-plasmid double-gene expression carrier takes pCDFDuet-1 or pETDuet-1as a foundation and a formate dehydrogenase gene and a leucine dehydrogenase gene are inserted into a polycloning site; the double-plasmid co-expression system takes any two of the pCDFDuet-1, the pETDuet-1 and pET-28b as foundations and comprises the leucine dehydrogenase gene and the formate dehydrogenase gene. The double-gene co-expression system is constructed and is transformed to obtain recombination genetic engineering bacteria; fermented and cultured recombination genetic engineering bacterium living cells are used as a catalyst and are applied to a catalytic reaction system for preparing the L-2-aminobutyric acid; the production efficiency is high, a coenzyme does not need to be added in a reaction process and the cost is saved.

A yeast extract containing 0.2% or more of γ-Glu-Abu based on dry weight of the yeast extract is produced by culturing a yeast, such as Saccharomyces cervisiae or Candida utilis, in a medium containing a compound selected from Abu (L-2-aminobutyric acid) and γ-Glu-Abu (L-γ-glutamyl-L-2-aminobutyric acid), and preparing a yeast extract from the obtained cells.

The invention relates to a processing technique of L-2-amino butyramide hydrochloride. The processing technique includes: using 2-chlorobutyric acid as a raw material and hexamethylenetetramine as a catalyst to prepare 2-aminobutyric acid; using L-tartaric acid to resolve 2-aminobutyric acid to obtain L-2-aminobutyric acid, acrylating L-2-aminobutyric acid to obtain L-2-aminobutyryl compound, and obtaining L-2-amino butyramide hydrochloride under the condition of ammonia water. The processing technique has the advantages that by the processing technique, reaction yield is increased, and byproducts are few. In addition, the processing technique is mild in reaction condition, easy in reaction control, low in cost, high in yield, high in product purity, low in equipment requirement and suitable for industrial production, and technique safety is improved greatly.

The invention discloses a method for preparing D-2-aminobutyric acid or L-2-aminobutyric acid by a chemical resolution method, comprising the following steps: carrying out reaction between the DL-2-aminobutyric acid and the L-dihydroxysuccinic acid or D-dihydroxysuccinic acid in inorganic acid, cooling, precipitating the solid intermediate compound, recrystallizing the compound with water, carrying out ammonolysis reaction on the recrystallized intermediate compound and the ammonolysis agent, cooling, filtering, and washing to obtain D-2-aminobutyric acid or L-2-aminobutyric acid. The D-2-aminobutyric acid or L-2-aminobutyric acid prepared by the method in the invention has high purity and high yield, and the cost is substantially reduced, thus being applicable to industrial scale production.

The invention discloses a preparation method of an idelalisib intermediate, the idelalisib intermediate (S)-N-(2-Boc-aminobutyryl)-2-fluoro-6-N-phenyl benzamide is prepared from aniline, N-Boc-L-2-aminobutyric acid, DCC, triethylamine and 2-fluoro-5-nitrobenzoyl chloride as raw materials by a three-step reaction of condensation reaction, amidation reaction and ring closure reaction for one-pot synthesis, and the condensation reaction and the amidation reaction do not require separation. The preparation method is a preparation method with the advantages of high yield, low cost, less waste, easyoperation and suitable industrialization.