caa a22 4500 606212981 CHVBK 20210128101045.0 cr unu---uuuuu 210128e20150501xx s 000 0 eng 10.1007/s10295-015-1604-y doi (NATIONALLICENCE)springer-10.1007/s10295-015-1604-y Expression of a novel epoxide hydrolase of Aspergillus usamii E001 in Escherichia coli and its performance in resolution of racemic styrene oxide [Elektronische Daten] [Die Hu, Cun-Duo Tang, Biao Yang, Jia-Chi Liu, Tao Yu, Chao Deng, Min-Chen Wu] 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 1.44U/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 Hg2+, Sn2+, Cu2+, Fe3+ and Zn2+. The K m and V max of re-AuEH2 were 5.90mM and 20.1U/mg towards (R)-SO, while 7.66mM and 3.19U/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. Society for Industrial Microbiology and Biotechnology, 2015 Aspergillus usamii nationallicence Epoxide hydrolase nationallicence Styrene oxide nationallicence Enantioselectivity nationallicence Molecular docking simulation nationallicence Hu Die Key Laboratory of Carbohydrate Chemistry and Biotechnology, School of Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, Jiangsu, People's Republic of China aut Tang Cun-Duo China-UK-NYNU-RRes Joint Laboratory of Insect Biology, Nanyang Normal University, 1638 Wolong Road, 473061, Nanyang, Henan, People's Republic of China aut Yang Biao Wuxi Medical School, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, Jiangsu, People's Republic of China aut Liu Jia-Chi Wuxi Medical School, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, Jiangsu, People's Republic of China aut Yu Tao School of Pharmaceutical Science, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, Jiangsu, People's Republic of China aut Deng Chao Wuxi Medical School, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, Jiangsu, People's Republic of China aut Wu Min-Chen Wuxi Medical School, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, Jiangsu, People's Republic of China aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/5(2015-05-01), 671-680 1367-5435 42:5<671 2015 42 10295 https://doi.org/10.1007/s10295-015-1604-y text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10295-015-1604-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hu Die Key Laboratory of Carbohydrate Chemistry and Biotechnology, School of Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, Jiangsu, People's Republic of China aut NATIONALLICENCE 700 1- Tang Cun-Duo China-UK-NYNU-RRes Joint Laboratory of Insect Biology, Nanyang Normal University, 1638 Wolong Road, 473061, Nanyang, Henan, People's Republic of China aut NATIONALLICENCE 700 1- Yang Biao Wuxi Medical School, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, Jiangsu, People's Republic of China aut NATIONALLICENCE 700 1- Liu Jia-Chi Wuxi Medical School, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, Jiangsu, People's Republic of China aut NATIONALLICENCE 700 1- Yu Tao School of Pharmaceutical Science, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, Jiangsu, People's Republic of China aut NATIONALLICENCE 700 1- Deng Chao Wuxi Medical School, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, Jiangsu, People's Republic of China aut NATIONALLICENCE 700 1- Wu Min-Chen Wuxi Medical School, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, Jiangsu, People's Republic of China aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/5(2015-05-01), 671-680 1367-5435 42:5<671 2015 42 10295