Biotechnology Letters · 2010 · 28 citations · 9 references
Life SciencesBiochemistryBiotechnologyBiocatalysisEnzymologyProtein EngineeringBiocatalyst EngineeringEnzyme FunctionEnzyme ProductionProtein Expression EngineeringYeast DisplayYeast Surface DisplayCell Surface DisplayEnzyme Expression In YeastEscherichia ColiGlycosylphosphatidylinositol AnchorsHydrolytic EnzymesYeast Strain EngineeringYeast Strain ImprovementYeast-substrate Interactions
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.
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Takeshi Matsumoto, Hideki Fukuda, Mitsuyoshi Ueda et al. · Applied and Environmental Microbiology · 2002 · 169 citations · Full text
Primary structure of the <i>Saccharomyces cerevisiae GAL7</i> gene
Tajima Masahiro, Yasuhisa Nogi, Toshio Fukasawa · Yeast · 1985 · 145 citations