Purification and characterization of chymotrypsin-like serine protease from Euphorbia rigida latex
Current Topics in Peptide and Protein Research, vol.18, pp.35-40, 2017 (Scopus)
- Publication Type: Article / Article
- Volume: 18
- Publication Date: 2017
- Journal Name: Current Topics in Peptide and Protein Research
- Journal Indexes: Scopus
- Page Numbers: pp.35-40
- Keywords: Characterization, Chymotrypsin-like serine protease, Euphorbia rigida, Latex, Purification
- Bursa Uludag University Affiliated: No
Abstract
A chymotrypsin-like serine protease was purified from Euphorbia rigida plant latex using 50% acetone precipitation. Enzyme activity was determined using protease substrates and inhibitors. The purified enzyme preparate showed the highest activity for the chromogenic substrate SucAAPFpNA, indicating a chymotrypsin-like activity. The partially purified enzyme was observed as a single band on sodium dodecyl sulfate poliacrylamide gel electrophoresis (SDS-PAGE) and its relative molecular weight was estimated as 66.4 kDa. The pH optima of the enzyme was found to be at pH 8 and the enzyme was stable over a wide range of pH. The thermal optima of the enzyme was found to be at 60°C and the enzyme was stable up to 60 °C after 1 hour of preincubation. Proteolytic activity was strongly inhibited by a serine protease-specific inhibitor PMSF. HgCl2 inhibited 51% of chymotrypsin activity. Overall, this enzyme may be considered as a strong candidate for various applications in the food and biotechnological industries due to its stability under harsh conditions.