Purification and characterization of a serine protease (CESP) from mature coconut endosperm

BMC Research Notes

Table 2 Hydrolysis of fluorogenic peptide substrate by the purified CESP

Fluorogenic Substrates	Concentration used (mM)	Relative Fluorescence Intensity
Boc-Leu-Arg-Arg-MCA	0.05	698.00
Boc-Val-Pro-Arg-MCA	0.05	583.50
Suc-Ala-Ala-Phe-MCA	0.05	120.70
Suc-Leu-Leu-Val-Tyr-MCA	0.05	0.060
Boc-Phe-Ser-Arg-MCA	0.05	625.85
Bz-Phe-Val-Arg-βNA	0.05	422.90
Bz-Leu-Leu-Glu-βNA	0.05	0.020

100 μl of reaction mixture containing 10 μl of purified enzyme, 0.05 mM of the substrates and 100 mM Tris HCl pH 8.5 was incubated at 37°C for 30 min and the reaction was stopped by the addition of 1.9 ml of ethanol. The product of hydrolysis was measured spectroflourimetrically (for AMC-Exc-380 nm, Em-460 nm and for βNA-Exc-335 nm, Em-410 nm).

ISSN: 1756-0500