title: Functional expression and magnetic nanoparticle-based Immobilization of a protein-engineered marine fish epoxide hydrolase of Mugil cephalus for enantioselective hydrolysis of racemic styrene oxide.
abstract: A triple-point mutated fish microsomal epoxide hydrolase (mEH) gene from Mugil cephalus was expressed in Escherichia coli in the presence of various chaperones to prevent protein aggregations. The enantioselective hydrolytic activity was more than doubled by co-expressing the EH mutant gene with pGro7 plasmid. The highly active EH mutant with a his-tag was immobilized onto magnetic silica assembled with NiO nanoparticles. The immobilized mEH mutant was re-used more than 10 times with less than 10% activity loss. (S)-Styrene oxide with 98% enantiopurity was repeatedly obtained with over 50% of the theoretical yield by the magnetically separable high-performance mEH mutant.
authors.affiliation: Department of Food Science and Biotechnology, Kyungsung University, Busan 608-736, South Korea.
authors.forename: Sung Hee
authors.initials: SH
authors.lastname: Choi
authors.forename: Hee Sook
authors.initials: HS
authors.lastname: Kim
authors.forename: In Su
authors.initials: IS
authors.lastname: Lee
authors.forename: Eun Yeol
authors.initials: EY
authors.lastname: Lee
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity; D004800:Enzymes, Immobilized / Q000378:metabolism; D004851:Epoxide Hydrolases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D004852:Epoxy Compounds / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006868:Hydrolysis; D058185:Magnetite Nanoparticles; D008861:Microsomes / Q000201:enzymology*; D050505:Mutant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D010957:Plasmids; D017354:Point Mutation; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D023701:Smegmamorpha
chemical_list: D004800:Enzymes, Immobilized; D004852:Epoxy Compounds; D058185:Magnetite Nanoparticles; D050505:Mutant Proteins; D011993:Recombinant Fusion Proteins; C013690:styrene oxide; D004851:Epoxide Hydrolases