Affinity Purification And Immobilization Of Chitinase From Bacillus Sp.R2

Bacillus sp. R2 chitinase was purified to homogeneity using ammonium sulphate precipitation at 80% saturation, affinity chromatography on swollen crab shell chitin column, and followed by gel filtration chromatography on Sephadex G-100; the enzyme was purified to 14.89-fold and yield of 14.77%, respectively.Furthermore the purified chitinase was subjected to three immobilization techniques on ten various solid carriers;1-physical adsorption immobilization on activated charcoal and silica gel carriers; 2-covalent binding on crab shell chitin and chitosan; 3Ionic binding on CM-Sepharose, Q-Sepharose, DEAE Cellulose, Amberlite IRC 50, Amberlite CG-120 (NA) and Amberlite CG -4B (OH)respectively Immobilization results revealed that covalent and ionic binding were the best techniques whereas chitosan, Amberlite IRC50 and chitin gave the highest immobilization yields 72.9, 70.26 and 63.53% with the highest activity yields 63.36, 58.26 and 53.03% respectively.These findings improve the effectiveness of swollen crab shell chitin as matrix for chitinase affinity purification whereas chitin and chitosan as active natural biopolymers for chitinase continuous production through immobilization. (C) 2015 Published by Elsevier Ltd.