Purification, characterization and the use of recombinant prolyl oligopeptidase from Myxococcus xanthus for gluten hydrolysis


PROTEIN EXPRESSION AND PURIFICATION, cilt.129, ss.101-107, 2017 (SCI İndekslerine Giren Dergi) identifier identifier identifier

  • Cilt numarası: 129
  • Basım Tarihi: 2017
  • Doi Numarası: 10.1016/j.pep.2016.09.016
  • Sayfa Sayıları: ss.101-107


Prolyl oligopeptidase (POP, EC is a cytosolic serine protease that hydrolyses proline containing small peptides. The members of prolyl oligopeptidase family play important roles in many physiological processes such as neurodegenerative diseases, maturation and degradation of peptide hormones. Thus the enzyme has been purified and characterized from various sources to elucidate the potential use as therapeutics. In this study recombinant Myxococcus xanthus prolyl oligopeptidase expressed in E. colt was purified 60.3 fold, using metal-chelate affinity and gel permeation chromatography. The recombinant enzyme had a monomeric molecular weight of 70 kDa. Isoelectric point of the enzyme was found to be approximately 6.3 by two-dimensional polyacrylamide gel electrophoresis. The optimum pH and temperature was estimated as 7.5 and 37 degrees C, respectively. The purified enzyme was stable in a pH range of 6.0 -8.5 and thermally stable up to 37 degrees C. The K-m and V-max values were 0.2 mM and 3.42 mu mol/min/mg. The proteolytic activity was inhibited by active-site inhibitors of serine protease, Z-Pro-Prolinal, PMSF, and metal ions, Cd2+, and Hg2+. Furthermore, the hydrolysis efficiency of the recombinant prolyl oligopeptidase was investigated with wheat gluten. (C) 2016 Elsevier Inc. All rights reserved.