10 results about "Halohydrin dehalogenase" patented technology

The invention discloses a halohydrin dehalogenase mutant and application of the halohydrin dehalogenase mutant in preparation of (R)-4-phenyl-2-oxazolidinone, and belongs to the technical field of enzyme engineering and chemical engineering, amino acid residues of wild type halohydrin dehalogenase are mutated, and the halohydrin dehalogenase mutant with high enzyme activity and high stereoselectivity is screened out. The halohydrin dehalogenase mutant provided by the invention has high enzyme activity and high stereoselectivity in the application of catalyzing (S)-2-phenyl ethylene oxide to be converted into (R)-4-phenyl-2-oxazolidinone, the catalytic efficiency of the mutant T126L / F170M can reach 1.7 times of that of wild halohydrin dehalogenase, and the halohydrin dehalogenase mutant has extremely high industrial application value.

The invention discloses a halohydrin dehalogenase mutant and its application in preparing (R)-4-phenyl-2-oxazolidinone, belongs to the field of enzyme engineering and chemical engineering technology, the amino acid residues of wild-type halohydrin dehalogenase are mutated, and halohydrin dehalogenase mutants with high enzyme activity and high stereoselectivity are screened out. The halohydrin dehalogenase mutant provided by the present invention has high enzyme activity and high stereoselectivity in the application of catalyzing (S)-2-phenyloxirane to convert (R)-4-phenyl-2-oxazolidinone, and the catalytic efficiency of mutant T126L / F170M can reach 1.7 times that of wild-type halohydrin dehalogenase, with extremely high industrial application value.

The invention belongs to the field of biotechnology, and discloses a halohydrin dehalogenase mutant and encoding gene, recombinant vector, recombinant strain, enzyme preparation and their application in synthesizing chiral (S)-5-phenoxymethyl-2-oxazolidinone compounds. The halohydrin dehalogenase mutant is a combination mutation of an enzyme with the amino acid sequence shown in SEQ ID NO.1 at positions 15, 137 and 179. The mutant can significantly improve the ee value of catalyzing phenyl glycidyl ether to synthesize chiral (S)-5-phenoxymethyl-2-oxazolidinone compounds, and the purity of catalytic synthesis and splitting is higher, which is conducive to industrial production.

The invention discloses a method for improving the biosynthesis efficiency of epichlorohydrin, which utilizes immobilized halohydrin dehalogenase to catalytically synthesize epichlorohydrin, and utilizes modified strong base anion exchange resin to adsorb chloride ions generated in the catalytic synthesis of epichlorohydrin, thereby solving the problem of reaction balance caused by chloride ion accumulation and improving the biosynthesis efficiency of epichlorohydrin. The synthesis efficiency of a target product is improved.

The invention discloses an epoxide hydrolase mutant and a construction method and application of a co-immobilized enzyme of the epoxide hydrolase mutant. The inventor firstly constructs a recombinant expression vector of epoxide hydrolysis MdEH, then constructs a recombinant expression vector of point mutation MdEH-K71I on the basis, then constructs an MdEH-K71I recombinant expression vector suitable for being used in bacillus subtilis, and then replaces a promoter and a signal peptide in the expression vector. The modified recombinant expression vector is used for transforming bacillus subtilis, so that efficient secretory expression of the epoxide hydrolase in a bacillus subtilis expression system is realized. The process conditions of co-immobilization of epoxide hydrolase and halohydrin dehalogenase are further explored, ECR8285 resin is selected as an immobilization carrier, and the conversion rate of glycerin generated by hydrolysis finally reaches 80.29%.

The present application discloses a halohydrin dehalogenase mutant, its encoding gene, a recombinant plasmid containing the encoding gene, a genetically engineered bacterium containing the recombinant plasmid, and its application, belonging to the field of genetic engineering. The halohydrin dehalogenase mutant of the present application is based on the amino acid sequence of the original halohydrin dehalogenase shown in SEQ ID NO.1, and its amino acid sequence is subjected to the following combined mutations: asparagine at position 179 is mutated to leucine; threonine at position 197 is mutated to alanine; leucine at position 198 is mutated to tyrosine; serine at position 224 is mutated to alanine; and lysine at position 227 is mutated to arginine. The combined mutations of the present application can produce targeted changes in the amino acid sequence, improve the catalytic activity and stereoselectivity of the halohydrin dehalogenase mutant, and at the same time streamline the number of mutation sites and omit specific transformation operations, saving time, labor, and cost, and facilitating industrial production.

The invention relates to a directional transformation method for improving enzyme activity of halohydrin dehalogenase, and belongs to the technical field of bioengineering. According to a halohydrin dehalogenase mutant, single-point mutation or combined mutation is carried out on 60th alanine and 179th histidine in a sequence shown in SEQ ID NO.1 to obtain the mutant. Compared with wild halohydrin dehalogenase, the mutant obtained in the invention has obviously improved enzyme activity and dehalogenation efficiency in catalytic preparation of 1-chloro-2-propanol and 2-chloro-1-ethanol, the enzyme activity of the mutant is improved by 1.78 times, the dehalogenation efficiency is improved by 77.8%, and the mutant has a good industrial application prospect.

This invention relates to the fields of enzyme engineering and biocatalysis, disclosing a halohydrin dehalogenase mutant and its catalytic enantioselective dehalogenation hydroxylation reaction of a series of racemic ε-halohydrins, preparing corresponding chiral ε-diols and chiral ε-halohydrins. The invention utilizes a fungicide derived from *Agrobacterium radiodurans* (…). Agrobacterium tumefaciens The wild-type haloalcohol dehalogenase HheC (amino acid sequence SEQ ID NO:2) of AD1 was used as the parent enzyme, and was obtained by combining 2 to 6 amino acid site mutations at L142, W249, P84, T134, N176, and F186. The mutant can be used to catalyze the kinetic resolution and dehalogenation reactions of racemic ε-haloalcohols, synthesizing a series of high-optical-purity chiral ε-diols and chiral ε-haloalcohols. These compounds are important chiral building blocks for the preparation of various pharmaceutical chemicals and have broad industrial application value.

This application discloses a method for targeted modification to improve the activity of halohydrin dehalogenase, belonging to the field of bioengineering technology. The halohydrin dehalogenase mutant obtained by this application is produced by single-point mutation or combination mutation of alanine at position 60 and histidine at position 179 in the sequence shown in SEQ ID NO.1. Compared with the wild-type halohydrin dehalogenase, the mutant obtained in this application shows significantly improved enzyme activity and dehalogenation efficiency in the catalytic preparation of 1-chloro-2-propanol and 2-chloro-1-ethanol. The mutant enzyme activity is increased by 1.78 times, and the dehalogenation efficiency is increased by 77.8%, showing good prospects for industrial application.

The invention relates to a halohydrin dehalogenase curing method, which comprises the following steps: dissolving halohydrin dehalogenase in a polyvinylpyrrolidone aqueous solution, and adding urea and formaldehyde to carry out curing reaction to obtain immobilized halohydrin dehalogenase. According to the method, the immobilized enzyme mass ratio of the halohydrin dehalogenase can be increased, the enzyme titer per unit mass can be increased, the enzymatic reaction yield can be increased, the immobilized product of the halohydrin dehalogenase is stable, the enzymatic reaction process is high in controllability, and the yield and purity of ATS-5 enzyme reaction can be increased.