Open AccessOPTIMIZATION, ISOLATION AND PARTIAL CHARACTERIZATION OF PROTEASES FROM UNDERUTILIZED AND COMMON FOOD LEGUMES
Author(s) -
. Anupama,
Malliga Marimuthu,
Uma Sundaram,
P. Gurumoorthi
Publication year - 2013
Publication title -
international research journal of pharmacy
Language(s) - English
Resource type - Journals
ISSN - 2230-8407
DOI - 10.7897/2230-8407.04722
Subject(s) - isolation (microbiology) , proteases , biology , characterization (materials science) , microbiology and biotechnology , food science , biochemistry , nanotechnology , materials science , enzyme
The present study aims at the isolation and partial characterization of proteases from six legume seeds, horse gram, jack bean, lima bean and velvet bean (white and black seeds). The extraction of these seeds was optimized using response surface methodology. The best buffer for extraction was seen to be 10 mM Tris HCl at pH 8 with 25 mg/ml-45 mg/ml protein release. The ideal extraction parameters exhibiting highest protease release (91.9112 U) were pH 4.86, 10-15˙C with 70 hours incubation time. The optimized extracts were subjected to (NH4)2SO4 precipitation to obtain crude enzyme solution. The proteolytic activity of the precipitated enzyme was again checked. The effect of pH and temperature of protease activity in the crude enzyme solution were also determined. The pH profile of proteases showed proteolytic activity at pH 7.5 to 9.0. The paper concludes that leguminous seeds can be source of proteases for industrial purposes