Cloning and Bioinformatics Analysis of Epoxide Hydrolase Gene from Aspergillus usamii

Abstract:

Epoxide hydrolase can be effectively used in the resolution of epoxides for producting optically active epoxides and vicinal diols. A gene encoding a novel epoxide hydrolase from Aspergillus usamii E001 was cloned by using reverse transcription PCR and newly constructed Thairpin structure-mediated PCR amplification(THSO-PCR) techniques. The cloned gene(named Aueh2) is 2,481 bp in length,harboring 5′ and 3′ flanking regulatory regions and the encoded cDNA sequence interrupted by six introns. The open reading frame of Aueh2 encodes a protein of 395-aa(designated AuEH2) with the calculated molecular weight of 44.6 kD. The primary structure analysis of AuEH2 demonstrated that it is a hydrophilic protein,and belongs to the α/β hydrolase fold family. The structure of AuEH2 displays that the catalytic center is situated between the α/β core domain and the lid domain with a catalytic triad consisting of Asp191,His369,and Glu343. The results will lay a foundation for further research of the AuEH2 and its application in depth.