Original Research Paper

Biotechnology Letters

, Volume 32, Issue 5, pp 655-659

First online:

Open Access This content is freely available online to anyone, anywhere at any time.

Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system

  • Takeshi FukudaAffiliated withDepartment of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui
  • , Kouta TsuchiyamaAffiliated withDepartment of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui
  • , Hirokazu MakishimaAffiliated withDepartment of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui
  • , Katsumi TakayamaAffiliated withDepartment of Chemistry and Biology Engineering, Fukui National College of Technology
  • , Ashok MulchandaniAffiliated withDepartment of Chemical and Environmental Engineering, University of California
  • , Kouichi KurodaAffiliated withDivision of Applied Life Sciences, Graduate School of Agriculture, Kyoto University
  • , Mitsuyoshi UedaAffiliated withDivision of Applied Life Sciences, Graduate School of Agriculture, Kyoto University
  • , Shin-ichiro SuyeAffiliated withDepartment of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui Email author 

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.

Keywords

Cell surface engineering Flo1p anchor system GPI anchor system Organophosphorus hydrolase