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Volume 148, Issue 9

Cloning and expression of the phosphotriesterase gene from C11

bThe GenBank accession number for the gene is AF469117.

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Abstract

The cloning of a gene encoding the novel phosphotriesterase from C11, which enabled it to use the organophosphate (OP) coroxon as its sole phosphorus source, is described. The gene, called (ydrolysis f oroxon) consists of 501 bp and encodes a protein of 19 kDa. This protein had no sequence similarity to any proteins in the SWISS-PROT/GenBank databases. When a spectinomycin-resistance cassette was placed in this gene, phosphotriesterase activity was abolished and C11 could no longer grow with coroxon as the sole phosphorus source. Overexpression and purification of HocA as a maltose-binding protein fusion produced a protein having a broad substrate specificity across oxon and thion OPs. Michaelis–Menten kinetics were observed with the oxon OPs, but not with the thion OPs. End-product inhibition was observed for coroxon-hydrolytic activity. Increased expression of was observed from an integrative fusion when cultures were grown in the absence of phosphate, suggesting that it might be part of the Pho regulon, but the phosphate-regulated promoter was not cloned in this study. This is believed to be the first study in which a gene required for an organism to grow with OP pesticides as a phosphorus source has been isolated.

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Keyword(s): coroxon , MBP, maltose-binding protein , OP, organophosphate and organophosphate hydrolase
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2002-09-01
2024-04-16
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Cloning and expression of the phosphotriesterase gene hocA from Pseudomonas monteilii C11bbThe GenBank accession number for the hocA gene is AF469117.
Microbiology 148, 2687 (2002); https://doi.org/10.1099/00221287-148-9-2687
/content/journal/micro/10.1099/00221287-148-9-2687
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