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Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system |
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| Authors: | Takeshi Fukuda Kouta Tsuchiyama Hirokazu Makishima Katsumi Takayama Ashok Mulchandani Kouichi Kuroda Mitsuyoshi Ueda Shin-ichiro Suye |
| Institution: | (1) Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, 3-9-1, Bunkyo, Fukui 910-8507, Japan;(2) Department of Chemistry and Biology Engineering, Fukui National College of Technology, Geshi-cho, Sabae, Fukui 916-8507, Japan;(3) Department of Chemical and Environmental Engineering, University of California, Riverside, CA 92521, USA;(4) Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa, Sakyo-ku, Kyoto 606-8502, Japan; |
| Abstract: | Organophosphorus hydrolase (OPH) hydrolyzes organophosphorus esters. We constructed the yeast-displayed OPH using Flo1p anchor system. In this system, the N-terminal region of the protein was fused to Flo1p and the fusion protein was displayed on the cell surface. Hydrolytic reactions with paraoxon were carried out during 24 h of incubation of OPH-displaying cells at 30°C. p-Nitrophenol produced in the reaction mixture was detected by HPLC. The strain with highest activity showed 8-fold greater OPH activity compared with cells engineered using glycosylphosphatidylinositol anchor system, and showed 20-fold greater activity than Escherichia coli using the ice nucleation protein anchor system. These results indicate that Flo1p anchor system is suitable for display of OPH in the cell surface-expression systems. |
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