Generation of a mutagenized organophosphorus hydrolase for the biodegradation of the organophosphate pesticides malathion and demeton-S.

Journal of Applied Microbiology
D A Schofield, A A DiNovo

Abstract

The bacterial organophosphorus hydrolase (OPH) enzyme hydrolyses and detoxifies a broad range of toxic organophosphate pesticides and warfare nerve agents by cleaving the various phosphorus-ester bonds (P-O, P-F, P-CN, P-S); however, OPH hydrolyses these bonds with varying efficiencies. The aim of this study was to generate a variant OPH enzyme with improved hydrolytic efficiency against the poorly hydrolysed P-S class of organophosphates. The gene encoding OPH was sequentially mutated at specific codons by saturation mutagenesis and screened for improved activity against the P-S substrates demeton-S methyl and malathion. Escherichia coli lysates harbouring the variants displayed up to 177- and 1800-fold improvement in specific activity against demeton-S methyl and malathion, respectively, compared to the wild-type lysates. The specificity constants of the purified variant proteins were improved up to 25-fold for demeton-S methyl and malathion compared to the wild-type. Activity was associated with organophosphate detoxification as the hydrolysed substrate lost the ability to inhibit acetylcholinesterase. The improved hydrolytic efficiency against demeton-S translated to the improved ability to hydrolyse the warfare agent VX. O...Continue Reading

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Citations

May 16, 2014·Biochemical and Biophysical Research Communications·Young-Su JeongHak-Sung Kim
Feb 3, 2016·Environmental Science and Pollution Research International·Pauline JacquetEric Chabrière
Dec 15, 2015·Chemical Reviews·Yoon Jeong JangDavid G Churchill
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May 9, 2012·Biochimica Et Biophysica Acta·Andrew N Bigley, Frank M Raushel
Oct 27, 2018·Journal of Pesticide Science·Karla Alejo-GonzálezRafael Vazquez-Duhalt
Aug 20, 2020·Annals of the New York Academy of Sciences·Priya KatyalJin Kim Montclare
Sep 18, 2013·ACS Chemical Biology·Izhack ChernyDan S Tawfik
Jun 25, 2013·Journal of the American Chemical Society·Andrew N BigleyFrank M Raushel

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