The Bioinformatic Analysis,Heterologous Expression and Catalytic Activity of Epoxide Hydrolase from Bacillus sp.
Epoxide hydrolases are a class of enzymes that can catalyze the hydrolysis of epoxides to produce corre-sponding ortho-diols.The reactions catalyzed by epoxide hydrolases do not require coenzymes and metal ions with substrate specificity.It has a wide application values in the preparation of pharmaceutical intermediates.Epoxide hydrolases from Bacillus sp.V3-13 contains 383 amino residues with a theoretical molecular weight of 44 119.61 Da and an isoelectric point of 5.33.It also has an instability index of 39.25 and a grand average hydropathicity of-0.493.Therefore,it is stable and hydrophilic.The enzyme has no signal peptide,disulfide bonds and two conserva-tive domains named EHN(pfam06441)and MenH(COG0596).Using pET-28a(+)as vector and E.coli BL21(DE3)as host,the enzyme is successfully expressed at 30 ℃,0.4 mmol·L-1 IPTG and the induction time of 10 h.Recombiant EH with electrophoretical purity is obtained after purification by nickel column affinity chromato-graphy.The hydrolysis rates ofbenzyl glycidyl ether,3-(1-naphthoxy)-1,2-propylene oxide and styrene oxide cata-lyzed by the recombinant EH within 20 min is 14.40％,11.05％and 8.17％,respectively.
epoxide hydrolaseexpression and purificationepoxidebioinformatic analysis
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