The purification and properties of phosphonoacetate hydrolase, a novel carbon-phosphorus bond-cleavage enzyme from Pseudomonas fluorescens 23F

Eur J Biochem. 1995 Nov 15;234(1):225-30. doi: 10.1111/j.1432-1033.1995.225_c.x.

Abstract

A novel, inducible, carbon-phosphorus bond-cleavage enzyme, phosphonoacetate hydrolase, was purified from cells of Pseudomonas fluorescens 23F grown on phosphonoacetate. The native enzyme had a molecular mass of approximately 80 kDa and, upon SDS/PAGE, yielded a homogenous protein band with an apparent molecular mass of about 38 kDa. Activity of purified phosphonoacetate hydrolase was Zn2+ dependent and showed pH and temperature optima of approximately 7.8 and 37 degrees C, respectively. The purified enzyme had an apparent Km of 1.25 mM for its sole substrate 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase
  • Amino Acid Sequence
  • Carbon / metabolism*
  • Enzyme Activation
  • Enzyme Stability
  • Hydrogen-Ion Concentration
  • Hydrolysis
  • Metals / pharmacology
  • Molecular Sequence Data
  • Phosphoric Monoester Hydrolases / antagonists & inhibitors
  • Phosphoric Monoester Hydrolases / isolation & purification*
  • Phosphoric Monoester Hydrolases / metabolism
  • Phosphorus / metabolism*
  • Pseudomonas fluorescens / enzymology*
  • Substrate Specificity
  • Temperature

Substances

  • Metals
  • Phosphorus
  • Carbon
  • Alkaline Phosphatase
  • Phosphoric Monoester Hydrolases
  • phosphonoacetate hydrolase