61 results about "Epoxide Hydrolases" patented technology

The invention discloses methods of using cis-epoxyeicosantrienoic acids (“EETs”), inhibitors of soluble epoxide hydrolase (“sEH”), or a combination of an EET and an inhibitor of sEH, to reduce or prevent niacin-induced cutaneous vasodilation (“flushing”) in subjects suffering from this undesirable side effect of receiving therapeutic amounts of niacin.

The invention discloses a kidney bean epoxide hydrolase mutant with improved catalytic activity and enantioconvergent property, belonging to the fields of genetic engineering and protein expression. A strain of epoxide hydrolase mutant PvEH1 <L105I/ M160A/ M175I> with improved catalytic activity and enantio-convergent property can be obtained by reforming the molecular structure of epoxide hydrolase (EH) by means of a site-directed mutation biotechnology on the basis of EH with certain enantio-convergent catalysis characteristic. The mutant has the advantages of high enantio-selectivity and high catalytic activity, thus having greater application potential and also laying a theoretical foundation for the study of the EH.

The invention discloses an epoxide hydrolase sourced from Phaseolus vulgaris and an application thereof and belongs to the technical field of biology. The invention discloses an epoxide hydrolase gene (pveh2) sourced from a leguminous plant, Phaseolus vulgaris, as well as a corresponding amino sequence. The invention provides a recombinant plasmid pET28a(+)-pveh2 and a genetic engineering bacteria E.coli BL21(DE3)/pET28a(+)-pveh2, which comprise the gene. A recombinant epoxide hydrolase is prepared through expression by the bacterial strain. The invention provides a method of biocatalysis through fermentation with an enzyme produced by the recombinant bacterial strain constructed in the invention, thus producing enantiomerically pure R-type styrene oxide (ee > 99%); in addition, final yield can reach 49.3%, which is approximate to a theoretical value, 50%.

The invention discloses an epoxide hydrolase mutant as well as a gene and an application of the epoxide hydrolase mutant. The epoxide hydrolase mutant is protein which is obtained by substituting one amino acid at one site or multiple sites of the 123th, 128th, 144th 145th, 168th, 219th and 221st amino acid of the protein with an amino acid sequence as shown in SEQ ID No: 1 in a sequence table and deriving from the protein with the amino acid sequence as shown in SEQ ID No: 1 and has epoxide hydrolase activity. Compared with wild type epoxide hydrolase, the epoxide hydrolase mutant provided by the invention can greatly improve hydrolytic activity and enantioselectivity of the racemization epoxide, and is particularly suitable for splitting and catalyzing the racemization naphthyl glycidyl ether to prepare (S)- naphthyl glycidyl ether and further synthesize racemic medicament (S)-propranolol. The hydrolase mutant has good industrial application prospect.

The invention provides compositions comprising an alkene epoxidase and a selective epoxide hydrolase, such as a recombinant microorganism comprising a first heterologous nucleic acid encoding an alkene epoxidase and a second heterologous nucleic acid encoding a selective epoxide hydrolase. Exemplary alkene epoxidases include StyAB, while exemplary selective epoxide hydrolases include epoxide hydrolases from Sphingomonas, Solanum tuberosum, or Aspergillus. The invention also provides non-toxic methods of making enantiomerically pure vicinal diols or enantiomerically pure alpha-hydroxy carboxylic acids using these compositions and microorganisms.

The invention relates to a preparation method of (R)-propene carbonate, which comprises the following steps of: (1) mixing propylene oxide and epoxide hydrolase ECU1040 with water, carrying out reaction for 10-40 hours at the temperature of 0-5 DEG C, then, stopping the reaction, and distilling (R)-propylene oxide at normal atmosphere; and (2) mixing the obtained (R)-propylene oxide with an organic solvent, adding quinine and organic amine, leading in carbon dioxide gas at the temperature of 10-30 DEG C while stirring until the carbon dioxide gas is not absorbed, carrying out reaction for a total reaction time of 15-30 hours, and then obtaining the (R)-propene carbonate. The preparation method is simple and practical, the sources of the raw materials are wide, the reaction is carried out at the normal pressure, the risk is low, the equipment investment is small, the yield is high, the optical purity of the product reaches over 99% e.e, and thus, the preparation method has an excellentindustrial prospect.

The invention discloses an epoxide hydrolase from rhodosporidium paludigenum and application of the epoxide hydrolase, and belongs to the technical field of biology. The epoxide hydrolase and the application have the advantages that epoxide hydrolase genes (rpeh1) from the rhodosporidium paludigenum R. paludigenum and corresponding amino acid sequences are provided; recombinant plasmids pET28a(+)-rpe1 with the genes and genetically engineered bacteria E. coli BL21 (DE3)/pET28a(+)-rpeh1 are constructed, and recombinant epoxide hydrolases are prepared by the aid of bacterial strains; biologicalcatalysis can be carried out by the constructed genetically engineered bacteria, and accordingly the ee of enantiomerically pure R-type phenyl glycol obtained by means of preparation is higher than 73%.

The invention discloses a method for degrading 1,2,3-TCP (trichloropropane) by engineered strain bacillus subtilis. According to the method, firstly, alkyl halide dehalogenase genes, halogenohydrin dehalogenase genes and epoxide hydrolase genes which can degrade 1,2,3-TCP and intermediate products of the 1,2,3-TCP are synthesized; the gene sequence and the codase amino acid sequence are respectively shown as SEQ ID NO.1 to 6; then, the bacillus subtilis integrating carrier containing the gene is built; the carrier converts the bacillus subtilis; by a gene ectopic integration and resistance gene knock-out method, the gene is integrated into the bacillus subtilis genome; the engineered strain bacillus subtilis capable of secreting and expressing alkyl halide dehalogenase, halogenohydrin dehalogenase and epoxide hydrolase is obtained. The engineered strain bacillus subtilis has a good degradation effect on TCP and TCP intermediate products; the complete TCP degradation can be realized; the characteristics of high degradation efficiency, high TCP tolerance, stable inheritance and the like are realized; the application prospects are good.

The invention belongs to the technical field of new function materials, and particularly relates to magnetic cellulose-dimethyl imidazole zinc salt and a preparing method and application thereof. Cellulose is dissolved in NaOH-Urea aqueous solution, nanometer Fe3O4 is added while stirring, water is added for regeneration, and product separation and washing are conducted to obtain magnetic cellulose; the magnetic cellulose and zinc nitrate solution are mixed and stirred for 5-30 min, then 2-methylimidazole solution is added for reaction lasting 0.5-24 h while stirring, and the obtained solid is subjected to magnetic separation, washing and drying to obtain the magnetic cellulose-dimethyl imidazole zinc salt. The product can be used as a carrier for enzyme immobilization, can immobilize a series of enzymes including lipase, papain, mung bean epoxide hydrolase and peroxidase, and has the potential to be a carrier of protein drugs.

The invention discloses a method for detecting soluble epoxide hydrolase by liquid chromatography-mass spectrometry. The method comprises the following steps: (1) screening a specific polypeptide of the soluble epoxide hydrolase from tissues, and showing the amino acid sequence of the specific polypeptide as SEQ ID NO.1; taking specific polypeptide as a standard substance, carrying out detecting by utilizing a mass spectrum isotope internal standard quantitative method and a liquid chromatography-mass spectrometry method, and establishing a standard curve by taking the concentration of the standard substance as an abscissa and the ratio of a quantitative daughter ion peak area to an isotope internal standard peak area as an ordinate; and (2) determining the concentration of the specific polypeptide in a sample by the liquid chromatography-mass spectrometry method, and then carrying out converting and calculating to obtain the concentration of the soluble epoxide hydrolase in the sample. According to the present invention, a peptide fragment with characteristics of stable enzymolysis and high enzymolysis efficiency is selected from the representative characteristic peptide fragmentwith stable property as the quantitative peptide fragment, such that advantages of accuracy, precision, sensitivity and the like are realized, and the sEH in the rat tissue can be effectively, accurately and quantitatively analyzed.

The invention discloses a method for preparing (R)-(4-chlorophenyl)oxirane from recombinant escherichia coli, and belongs to the technical field of biology. According to the method, tomato-derived epoxide hydrolase is expressed in escherichia coli, and a genetically engineered bacterium E.coli/sleh1 is constructed, and expression of the strain is used for preparing recombinant epoxide hydrolase. The invention provides a method for biological catalysis by using enzyme produced by fermenting the recombinant strain constructed in the method. (R)-(4-chlorophenyl)oxirane (ee is larger than 98%) ofenantiopure is prepared by catalyzing 150 mM racemized (4-chlorophenyl)oxirane; besides, the final yield reaches 47.5% and is close to the theoretical value of 50%.

The invention relates to an epoxide hydrolase mutant and an application thereof in enantiomeric normalizing hydrolysis of epoxides and discloses a series of epoxide hydrolase mutants, which have veryhigh catalytic activity and stereo-selectivity. The invention also discloses the application of the epoxide hydrolase mutant. By using the epoxide hydrolase mutant for catalyzing an enantiomeric normalizing hydrolysis reaction of a racemized styrene oxide and a derivative thereof, enantiomeric normalizing degree is high, reaction conditions are gentle, the process is environment-friendly, and theproduct has high optical purity. The obtained mutation site has universality about other epoxide hydrolases, so that the epoxide hydrolase mutant has extensive application prospect.

The invention provides design and construction of AuEH2G218S and AuEH2S247Y mutant enzymes with high enantio-selectivity. The mutation sites are finally determined by sequence homology analysis, homologous modeling, molecular docking and dynamic simulation. Mutant genes obtained by a megaprimer RPC technology are named Aueh2G218S and Aueh2S247Y. The invention further discloses a method for construction and efficient expression of mutant engineering bacteria; and the prepared mutant enzymes have the characteristics of high enantio-selectivity and catalytic activity, and have relatively large industrial production potential.

The invention discloses a kidney bean epoxide hydrolase mutant with improved catalytic activity and belongs to the fields of gene engineering and protein expression. The mutant mutates valine at the 106th site into isoleucine on the basis of amino acid shown in SEQ ID NO.2. The catalytic activity of mutant enzyme PvEH1V106I obtained by shaking flask induction for 10h, disclosed by the invention, is 234.75U/g, which is 1.49 times that of wild type enzyme (157.15U/g).

The invention discloses an engineering strain for biosynthesizing mogrol by taking glucose as a substrate as well as construction and application of the engineering strain, and belongs to the technical field of biology. A chassis yeast engineering bacterium is constructed by enhancing squalene oxidation module expression, improving an MVA metabolic pathway and knocking out a lanosterol competition pathway, heterologous protein cucurbitadienol synthase, epoxide hydrolase and cell P450 enzyme are expressed on the basis of the chassis bacterium, and the yeast engineering bacterium for biosynthesis of mogrol is constructed. According to the method, mogrol can be synthesized from the beginning, the environment is not polluted, the yield is high, the cost is low, and a basis is provided for industrial production of mogrol.The finally produced mogrol reaches 1.2 g/L, which is the highest level reported at present.

N-benzylbenzamides that act as dual soluble soluble epoxide hydrolase (sEH)/peroxisome proliferator-activated receptor γ (PPARγ) modulators and are useful as medications in the treatment of Metabolic Syndrome (MetS) cluster diseases, including diabetes. Methods of making and using the same are further provided.

The invention discloses a mogroside synthesis regulation gene Sgesph7 and application thereof, and belongs to the field of biotechnology. According to the mogroside synthesis regulation gene Sgesph7 and the application thereof, by means of whole genome sequencing of Siraitia grosvenorii(Swingle)C.Jeffrey and the analysis of the RNA-seq sequence of different tissue, secondary metabolic pathways associated with the tissue specificity and characters are utilized, and therefore the Siraitia grosvenorii(Swingle)C.Jeffrey epoxide hydrolases Sgeph7 gene with Siraitia grosvenorii(Swingle)C.Jeffrey fruits specifically expressed; and the content of the mogroside in the Siraitia grosvenorii(Swingle)C.Jeffrey fruits can be regulated and controlled by adjusting the expression of the gene, and the geneis used for obtaining a new variety or new strain of the Siraitia grosvenorii(Swingle)C.Jeffrey with the high mogroside content and has important theoretical and practical significance in research ofthe taste mechanism of fruits of cucurbitaceae plants and the breeding work of the taste of the fruits of the cucurbitaceae plants.

Methods of using compounds that inhibit soluble epoxide hydrolase (sEH) for the treatment of ocular diseases, and in particular, of wet age-related macular degeneration (AMD), are disclosed herein.