Cell surface display of organophosphorus hydrolase in Pseudomonas putida using an ice-nucleation protein anchor.
A surface anchor system derived from the ice-nucleation protein (INP) from Pseudomonas syringe was used to localize organophosphorus hydrolase (OPH) onto the surface of Pseudomonas putida KT2440. Cells harboring the shuttle vector pPNCO33 coding for the INP-OPH fusion were capable of targeting OPH onto the cell surface as demonstrated by whole cell ELISA. The whole cell activity of P. putida KT2440 was shown to be 10 times higher than those of previous efforts expressing the same fusion protein in Escherichia coli. The capability of expressing enzymes on the surface of a robust and environmentally benign P. putida KT2440 should open up new avenues for a wide range of applications such as in situ bioremediation.
Shimazu
Mark
M
Department of Chemical and Environmental Engineering, University of California, Riverside, California 92521, USA.
Nguyen
Anne
A
Mulchandani
Ashok
A
Chen
Wilfred
W
D043303:Aryldialkylphosphatase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D001673:Biodegradation, Environmental; D002462:Cell Membrane / Q000201:enzymology*; D004789:Enzyme Activation; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D010575:Pesticides / Q000493:pharmacokinetics; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D016958:Pseudomonas putida / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012989:Soil Pollutants / Q000493:pharmacokinetics
D001425:Bacterial Outer Membrane Proteins; D008565:Membrane Proteins; D010575:Pesticides; D011993:Recombinant Fusion Proteins; D012989:Soil Pollutants; C063113:ice nucleation protein; D043303:Aryldialkylphosphatase