Purification and characterization of chymotrypsin-like serine protease from Euphorbia rigida latex
Makaleler > Tam Makale · 2017
Yazarlar: Ebru KOCADAĞ KOCAZORBAZ, Mehmet Ergüven, Mehmet Ergüven, Seda Suzan Memecan, Esra Menfaatli, Hasan YILDIRIM, Hasan YILDIRIM, Figen ZİHNİOĞLU
CURRENT TOPICS IN PEPTIDE AND PROTEIN RESEARCH, SCI Expanded İndekslerine Giren Dergi, Uluslararası, Scopus, 2017, cilt 18, ss 35-40
Özet
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.