Overexpression of parathion hydrolase in Escherichia coli stimulates the synthesis of outer membrane porin OmpF

Abstract

Parathion hydrolase (PH), also known as organophosphorus acid anhydrase, hydrolyses the triester linkage found in organophosphates including organophosphate pesticides and the nerve gas sarin. The enzyme is reported to be membrane-associated and the immature protein has a signal sequence of 29 amino acids. In experiments designed to examine the post-translational processing of the enzyme and to assess the distribution of the precursor and mature forms of the protein, we induced expression of the Flavobacterium balustinum PH structural gene, opd, in Escherichia coli strain BL21. Western blotting revealed that the induced PH was predominantly membrane-associated in E. coli but a protein band equivalent in size to mature PH was also found to be induced specifically in periplasmic fractions. This periplasmic protein was not PH, as it did not cross-react in Western blots, and N-terminal sequencing of the induced protein showed it to have 100% homology to the outer membrane protein OmpF.