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Purification and characterization of X-prolyl dipeptidyl peptidase fromLactobacillus casei subsp.casei LLG


X-Prolyl dipeptidyl peptidase, which hydrolysed X-Pro-Y almost specifically, has been purified to homogeneity from crude cell-free extracts ofLactobacillus casei subsp.casei LLG using fast protein liquid chromatography equipped with preparative and analytical anion exchange columns. The enzyme was purified to 274-fold by ammonium sulphate fractionation, and by two successive ion-exchange chromatographies with a recovery of 34%. The purified enzyme appeared as a single band on both native-polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulphate (SDS)-PAGE and had a molecular mass of 79 kDa. The pH and the temperature optima by the purified enzyme were 7.0 and 50°C, respectively. X-PDP was a serine-dependent enzyme, as both diisopropylfluorophosphate and phenylmethylsulphonylfluoride caused complete inhibition of the enzyme activity. The Michaelis constant (K m ) and maximum reaction velocity (V max ) values were 0.2 mm and 43 mm per milligram, respectively.

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Correspondence to B. H. Lee.

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Habibi-Najafi, M.B., Lee, B.H. Purification and characterization of X-prolyl dipeptidyl peptidase fromLactobacillus casei subsp.casei LLG. Appl Microbiol Biotechnol 42, 280–286 (1994).

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  • Sodium Dodecyl Sulphate
  • Anion Exchange
  • Ammonium Sulphate
  • Sodium Dodecyl Sulphate
  • Reaction Velocity