Figure 3
From: Purification and characterization of a serine protease (CESP) from mature coconut endosperm
Profile of BAPNA hydrolysis by the purified enzyme. (A) 50 μl of purified enzyme was assayed at pH 8.5 for BAPNA hydrolysis for different time periods up to 5 hrs using 50 mM BAPNA substrate (B) BAPNA hydrolysis was carried out for 1 hour with increasing volume of purified enzyme. (C) pH optimum of BAPNA hydrolysis by the purified CESP was studied in 100 mM buffers of varying pH. The buffers used were: Na Ac (5.1), MES (5.5, 6.0), PIPES (6.5, 7), Tris HCl (7.5 to 9) and CAPS (9.7, 10). (D) Measurement of Michaelis-Menten constant (Km): p-nitroaniline liberated was measured by varying substrate concentrations. Conditions of reactions are same as mentioned earlier. (E) SDS/PAGE zymogram analysis of the purified CESP, 2 μg (1st lane) & 3 μg (2nd lane) of the purified CESP after storage for one week.