title: Improved degradation of organophosphorus nerve agents and p-nitrophenol by Pseudomonas putida JS444 with surface-expressed organophosphorus hydrolase.
abstract: Pseudomonas putida JS444, isolated from p-nitrophenol (PNP) contaminated waste sites, was genetically engineered to simultaneously degrade organophosphorus pesticides (OP) and PNP. A surface anchor system derived from the ice-nucleation protein (INP) from Pseudomonas syringae was used to target the organophosphorus hydrolase (OPH) onto the surface of Pseudomonas putida JS444, reducing the potential substrate uptake limitation. Engineered cells were capable of targeting OPH onto the cell surface as demonstrated by western blotting, cell fractionation, and immunofluorescence microscopy. The engineered P. putida JS444 degraded organophosphates as well as PNP rapidly without instability problems associated with the engineered Moraxella sp. The initial hydrolysis rate was 7.90, 3.54, and 1.53 micromol/h/mg dry weight for paraoxon, parathion, and methyl parathion, respectively. The excellent stability in combination with the rapid degradation rate for organophosphates and PNP make this engineered strain an ideal biocatalyst for complete mineralization of organophosphates.
authors.affiliation: Department of Chemical and Environmental Engineeirng, University of California, Riverside, California 92521, USA.
authors.forename: Yu
authors.initials: Y
authors.lastname: Lei
authors.forename: Ashok
authors.initials: A
authors.lastname: Mulchandani
authors.forename: Wilfred
authors.initials: W
authors.lastname: Chen
mesh_terms: D043303:Aryldialkylphosphatase / Q000235:genetics / Q000378:metabolism*; D001673:Biodegradation, Environmental; D002619:Chemical Warfare Agents / Q000493:pharmacokinetics; D004787:Environmental Pollution / Q000517:prevention & control; D024861:Genetic Enhancement / Q000379:methods*; D007306:Insecticides / Q000493:pharmacokinetics*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009596:Nitrophenols / Q000493:pharmacokinetics*; D009943:Organophosphorus Compounds / Q000493:pharmacokinetics*; D015202:Protein Engineering / Q000379:methods; D016958:Pseudomonas putida / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D002619:Chemical Warfare Agents; D007306:Insecticides; D008565:Membrane Proteins; D009596:Nitrophenols; D009943:Organophosphorus Compounds; D011994:Recombinant Proteins; D043303:Aryldialkylphosphatase; C024836:4-nitrophenol