MRC Centre for Protein Engineering, Hills Road, Cambridge CB20QH, UK.
Protein Eng Des Sel. 2011 Jan;24(1-2):209-11. doi: 10.1093/protein/gzq046. Epub 2010 Jul 22.
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.
细菌磷酸三酯酶 (PTE) 以扩散限制的效率催化人工合成农药对氧磷的水解。在这里,我们描述了 PTE 变体 SS0.2 的选择和表征,该变体具有迄今为止描述的最高对氧磷酶周转率(kcat = 31,000 s-1)。PTE-SS0.2 是从一个结合位点突变体文库中通过一种新的筛选方法选择出来的,该方法结合了细菌菌落的部分裂解和荧光探针。