Directed Evolution of Methyl Parathion Hydrolase Based on the Multi-dimensional Features：Molecular Structure and Bioinformatics

doi:10.13560/j.cnki.biotech.bull.1985.2018-0221

Abstract

Abstract: As an environmental-friendly green catalyst,methyl parathion hydrolase can degrade organophosphorus pesticides such as methyl parathion remained in soil such,thus improving the hydrolysis efficiency of methyl parathion hydrolase becomes a well-concerned issue. Based on the multi-dimensional features such as structural and bioinformatics,9 key sites around the catalytic active center were determined. A screening library containing 1 500 mutants were constructed for directed evolution. After two rounds of screening,2 mutants with the improved catalytic efficiency,L258V and L273M were identified. Their catalytic efficiency constants k_cat/K_m values were 134.1（mmol/L）^-1·s^-1 and 166.5（mmol/L）^-1·s^-1,respectively,which increased by 27.3% and 58.1% compared with the wild type. This method of constructing a directed evolution library by semi-rational design not only improves the screening efficiency of methyl parathion hydrolase,but also provides a new idea to improve properties of enzyme.

Key words: methyl parathion hydrolase, directed evolution based on structure and bioinformation, active centre, evolutionary conservation, catalytic efficiency

REN Tian-lei, YANG Hai-quan, XU Fei. Directed Evolution of Methyl Parathion Hydrolase Based on the Multi-dimensional Features：Molecular Structure and Bioinformatics[J]. Biotechnology Bulletin, 2018, 34(10): 194-200.

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