Cloning, expression and enantioselective hydrolytic catalysis of a microsomal epoxide hydrolase from a marine fish, Mugil cephalus.
The cDNA of a marine fish microsomal epoxide hydrolase (mEH) gene from Mugil cephalus was cloned by rapid amplification of cDNA ends (RACE) techniques. The homology model for the mEH of M. cephalus showed a characteristic structure of alpha/beta-hydrolase-fold main domain with a lid domain over the active site. The characteristic catalytic triad, consisting of Asp(238), His(444), and Glu(417), was highly conserved. The cloned mEH gene was expressed in Escherichia coli and the recombinant mEH exhibited (R)-preferred hydrolysis activity toward racemic styrene oxide. We obtained enantiopure (S)-styrene oxide with a high enantiopurity of more than 99% enantiomeric excess and yield of 15.4% by batch kinetic resolution of 20 mM racemic styrene oxide.
Lee
Soo Jung
SJ
Department of Food Science and Biotechnology, Kyungsung University, Busan, Korea.
Kim
Hee Sook
HS
Kim
Sang Jin
SJ
Park
Sunghoon
S
Kim
Beum Jun
BJ
Shuler
Michael L
ML
Lee
Eun Yeol
EY
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D015153:Blotting, Western; D002384:Catalysis; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004851:Epoxide Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism; D005399:Fishes; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D006868:Hydrolysis; D008969:Molecular Sequence Data; D015394:Molecular Structure; D017433:Protein Structure, Secondary; D012623:Seawater; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013237:Stereoisomerism; D013379:Substrate Specificity
D018076:DNA, Complementary; D029941:Fish Proteins; D004851:Epoxide Hydrolases