State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Key Laboratory of Industrial Biotechnology, College of Life Sciences, Hubei University, Wuhan, China.
Mol Biotechnol. 2019 Nov;61(11):816-825. doi: 10.1007/s12033-019-00204-3.
Methyl parathion hydrolase (MPH) plays an important role in degrading a range of organophosphorus compounds. In order to display MPH on the cell surface of Escherichia coli strain RosettaBlue™, the Flavin-based fluorescent protein EcFbFP was severed as an auto-anchoring matrix. With net negative charges of EcFbFP supplying the driving forces, fusion protein MPH-EcFbFP through a two-step auto-surface display process was finally verified by (a) inner membrane translocation and (b) anchoring at outer membrane. Cells with surface-displayed MPH obtained activity of 0.12 U/OD600 against substrate methyl parathion. MPH when fused with engineered EcFbFP containing 20 net negative charges exhibited fivefold higher anchoring efficiency and tenfold higher enzymatic catalytic activity of 1.10 U/OD600. The above result showed that MPH was successfully displayed on cell surface and can be used for biodegradation of methyl parathion.
甲基对氧磷水解酶（MPH）在降解多种有机磷化合物方面发挥着重要作用。为了在大肠杆菌 RosettaBlue™菌株的细胞表面展示 MPH，黄素基荧光蛋白 EcFbFP 被用作自动锚定基质。带有净负电荷的 EcFbFP 提供驱动力，融合蛋白 MPH-EcFbFP 通过两步自动表面展示过程最终得到验证（a）内膜易位和（b）在外膜上的锚定。获得表面展示 MPH 的细胞对底物甲基对氧磷的活性为 0.12 U/OD600。当与含有 20 个净负电荷的工程 EcFbFP 融合时，MPH 的锚定效率提高了五倍，酶催化活性提高了十倍，达到 1.10 U/OD600。上述结果表明 MPH 已成功在细胞表面展示，并可用于甲基对氧磷的生物降解。
