Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system.
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.
Fukuda
Takeshi
T
Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, 3-9-1, Bunkyo, Fukui, 910-8507, Japan.
Tsuchiyama
Kouta
K
Makishima
Hirokazu
H
Takayama
Katsumi
K
Mulchandani
Ashok
A
Kuroda
Kouichi
K
Ueda
Mitsuyoshi
M
Suye
Shin-ichiro
S
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
D008565:Membrane Proteins; D009596:Nitrophenols; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D043303:Aryldialkylphosphatase; D010261:Paraoxon; C024836:4-nitrophenol