Enzymatic Hydrolysis of Organophosphates: Cloning and Expression of a Parathion Hydrolase Gene from Pseudomonas diminuta

摘要: Pseudomonas diminuta strain MG hydrolyzes parathion to diethylthiophosphoric acid and p-nitrophenol. The esterase responsible for this reaction is encoded by a gene located on plasmid termed pCMS1. was cloned into pBR322 the broad host range cloning vector pKT230. Enzyme activity detected in Escherichia coli strains that contained recombinant plasmids. A 1.5 kilobase BamHI fragment with single restriction sites SalI, PstI XhoI shown direct synthesis of enzyme. inserted high expression pUC7, resulting recombinant, pCMS40, used construct pKT230 derivatives containing hydrolase gene. Subsequent transfer plasmids cured derivative P. p-nitrophenol-utilizing resulted isolation transconjugants exhibited activity. highest enzyme observed MG.