THE PURIFICATION AND PROPERTIES OF PHOSPHONOACETATE HYDROLASE, A NOVEL CARBON-PHOSPHORUS BOND-CLEAVAGE ENZYME FROM PSEUDOMONAS-FLUORESCENS 23F

A novel, inducible, carbon-phosphorus bond-cleavage enzyme, phosphonoa cetate hydrolase, was purified from cells of Pseudomonas fluorescens 2 3F grown phosphonoacetate. The native enzyme had a molecular mass of a pproximately 80 kDa and, upon SDS/PAGE, yielded a homogenous protein b and with an apparent molecular mass of about 38 kDa. Activity of purif ied phosphonoacetate hydrolase was Zn2+ dependent and showed pH and te mperature optima of approximately 7.8 and 37 degrees C, respectively. The purified enzyme had an apparent K-m of 1.25 mM for its sole substr ate phosphonoacetate, and was inhibited by the structural analogues 3- phosphonopropionate and phosphonoformate. The NH2-terminal sequence of the first 19 amino acids displayed no significant similarity to other databank sequences.