Biotechnology and Bioengineering · 2001 · 154 citations · 36 references
Life SciencesBiotechnologyMicrobiologyEnzymologyMicrobial MetabolismSoil MicrobiologyEnzyme KineticsEnzyme Surface EngineeringMicrobial PesticidesPesticide BiotransformationPesticide ResiduesSoil EnzymologyHydrolysisMicrobial Degradation Of PesticidesMicrobial Pesticide DegradationOrganophosphorus PesticidesPesticide BiodegradationShuttle Vector ConstructionShuttle VectorsSubstrate Uptake
Moraxella sp., a native soil organism that grows on p-nitrophenol (PNP), was genetically engineered for the simultaneous degradation of organophosphorus (OP) pesticides and p-nitrophenol (PNP). The truncated ice nucleation protein (INPNC) anchor was used to target the pesticide-hydrolyzing enzyme, organophosphorus hydrolase (OPH), onto the surface of Moraxella sp., alleviating the potential substrate uptake limitation. A shuttle vector, pPNCO33, coding for INPNC-OPH was constructed and the translocation, surface display, and functionality of OPH were demonstrated in both E. coli and Moraxella sp. However, whole cell activity was 70-fold higher in Moraxella sp. than E. coli. The resulting Moraxella sp. degraded organophosphates as well as PNP rapidly, all within 10 h. The initial hydrolysis rate was 0.6 micromol/h/mg dry weight, 1.5 micromol/h/mg dry weight, and 9.0 micromol/h/mg dry weight for methyl parathion, parathion, and paraoxon, respectively. The possibility of rapidly degrading OP pesticides and their byproducts should open up new opportunities for improved remediation of OP nerve agents in the future.
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