Applied Biochemistry and Biotechnology

, Volume 31, Issue 1, pp 59–73

Detoxification of organophosphate pesticides using a nylon based immobilized phosphotriesterase fromPseudomonas diminuta

  • Steven R. Caldwell
  • Frank M. Raushel
Article

Abstract

A partially purified phophostriesterase was successfully immobilized onto nylon 6 and 66 membranes, nylon 11 powder, and nylon tubing. Up to 9000 U of enzyme activity was immobilized onto 2000 cm2 of a nylon 6 membrane where 1 U is the amount of enzyme necessary to catalyze the hydrolysis of 1.0 μmol of paraoxon/min at 25°C. The nylon 66 membrane-bound phosphotriesterase was characterized kinetically where the apparentKm value for the immobilized enzyme was 0.35 mM. This is 5-6 times higher than that observed for the soluble enzyme. However, nylon immobilization limited the maximum rate of paraoxon hydrolysis to less than 10% of the value measured for the soluble enzyme. The addition of the cosolvent, methanol, resulted in an increase in the apparentKm value for paraoxon hydrolysis but concentrations up to 40% had no negative effect on the catalytic effectiveness with the soluble or immobilized phosphotriesterase. Based on the kinetic analysis, methanol appears to be a competitive inhibitor for both forms of enzyme. The nylon powder immobilized enzyme was shown to be stable for at least 20 mo. The immobilization of the phosphotriesterase onto nylon provides a practical method for the detoxification of organophosphate pesticides.

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Copyright information

© Humana Press Inc. 1991

Authors and Affiliations

  • Steven R. Caldwell
    • 1
  • Frank M. Raushel
    • 2
  1. 1.Departments of BiochemistryTexas A&M University, College Station
  2. 2.Departments of ChemistryTexas A&M University, College Station

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