Expression of a novel epoxide hydrolase of Aspergillus usamii E001 in Escherichia coli and its performance in resolution of racemic styrene oxide

Journal of Industrial Microbiology & Biotechnology
Die HuMinchen Wu

Abstract

The full-length cDNA sequence of Aueh2, a gene encoding an epoxide hydrolase of Aspergillus usamii E001 (abbreviated to AuEH2), was amplified from the total RNA. Synchronously, the complete DNA sequence containing 5', 3' flanking regions, eight exons and seven introns was cloned from the genomic DNA. In addition, a cDNA fragment of Aueh2 encoding a 395-aa AuEH2 was expressed in Escherichia coli. The catalytic activity of recombinant AuEH2 (re-AuEH2) was 1.44 U/ml using racemic styrene oxide (SO) as the substrate. The purified re-AuEH2 displayed the maximum activity at pH 7.0 and 35 °C. It was highly stable at a pH range of 5.0-7.5, and at 40 °C or below. Its activity was not obviously influenced by β-mercaptoethanol, EDTA and most of metal ions tested, but was inhibited by Hg(2+), Sn(2+), Cu(2+), Fe(3+) and Zn(2+). The K m and V max of re-AuEH2 were 5.90 mM and 20.1 U/mg towards (R)-SO, while 7.66 mM and 3.19 U/mg towards (S)-SO. Its enantiomeric ratio (E) for resolution of racemic SO was 24.2 at 10 °C. The experimental result of re-AuEH2 biasing towards (R)-SO was consistent with the analytical one by molecular docking (MD) simulation.

References

Mar 20, 2002·Accounts of Chemical Research·Thomas C Bruice
Oct 9, 2002·Current Protein & Peptide Science·M Holmquist
Dec 14, 2005·Biochimica Et Biophysica Acta·Michael Kotik, Pavel Kyslík
Dec 17, 2008·Applied Microbiology and Biotechnology·Jung-Hee WooSang-Jin Kim
Jul 11, 2009·Applied Microbiology and Biotechnology·Won Jae Choi
Dec 7, 2011·Enzyme and Microbial Technology·Deniz YildirimOzlem Alptekin

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Citations

May 15, 2016·International Journal of Biological Macromolecules·Gabriel Stephani de OliveiraFelipe S Chambergo

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