Acetylsalicylate hydrolase of rabbit gastric mucosa. Isolation and purification

Academic Article

Abstract

  • The mechanism of hydrolysis of acetylsalicylate during absorption from the gastrointestinal tract has been investigated by identification, quantitation, and purification of a hydrolase from gastric mucosal homogenates. The hydrolase was found to be a soluble, cytosolic enzyme with a pH optimum in the slightly alkaline range, pH 8.6. Acetylsalicylate hydrolase activity was purified from the 100,000 g supernatant fraction by differential (NH4)2SO4 fractionation followed by DEAE-cellulose ion-exchange chromatography and Sephadex or Sephacryl gel filtration. The activity could also be fractionated on hydroxylapatite. The Sephadex-purified fraction containing peak enzyme activity gave a single protein band on SDS-polyacrylamide gel electrophoresis. The molecular weight of the acetylsalicylate hydrolase was 66,400 based on SDS-polyacrylamide gel electrophoresis of the Sephadex-purified enzyme and 59,000 based on gel filtration. By use of the technique described, acetylsalicylate hydrolase can be purified over 100-fold to a specific activity of 10.6 ╬╝mol x mg-1 x min-1.
  • Authors

    Published In

    Author List

  • Spenney JG; Nowell RM
  • Start Page

  • 215
  • End Page

  • 219
  • Volume

  • 7
  • Issue

  • 4