Protein Engineering Design and Selection · 2010 · 17 citations · 12 references
Life SciencesBiochemistryMicrobiologyBiocatalysisEnzymologyProtein EngineeringEnzyme KineticsEnzyme ScreeningMicrobial PesticidesPesticide BiotransformationPesticide ChemistryCell LysisDiffusion-limited GrowthMicrobial Degradation Of PesticidesMicrobial Pesticide DegradationPesticide Degradation KineticsSample Lysis
The bacterial phosphotriesterase (PTE) catalyses the hydrolysis of the man-made pesticide paraoxon with a diffusion-limited efficiency. Here we describe the selection and characterisation of PTE variant SS0.2 that possesses the highest paraoxonase turnover number so far described (k(cat) = 31,000 s⁻¹). The PTE-SS0.2 was selected from a library of binding-site mutants using a novel screening method that combines partial lysis of bacterial colonies and fluorogenic probes.
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