title: Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system.
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 degrees 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.
authors.affiliation: Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, 3-9-1, Bunkyo, Fukui, 910-8507, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Fukuda
authors.forename: Kouta
authors.initials: K
authors.lastname: Tsuchiyama
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Makishima
authors.forename: Katsumi
authors.initials: K
authors.lastname: Takayama
authors.forename: Ashok
authors.initials: A
authors.lastname: Mulchandani
authors.forename: Kouichi
authors.initials: K
authors.lastname: Kuroda
authors.forename: Mitsuyoshi
authors.initials: M
authors.lastname: Ueda
authors.forename: Shin-ichiro
authors.initials: S
authors.lastname: Suye
mesh_terms: D043303:Aryldialkylphosphatase / Q000235:genetics / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009596:Nitrophenols / Q000032:analysis; D010261:Paraoxon / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D013696:Temperature; D013997:Time Factors
chemical_list: D008565:Membrane Proteins; D009596:Nitrophenols; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D043303:Aryldialkylphosphatase; D010261:Paraoxon; C024836:4-nitrophenol