Abstract:
The activity of enzyme hydrolyzing colloidal chitin has been measured on extracts from a number of higher plants belonging to eleven families. Chitinase activity could be detected in dry seeds which increased during germination. Its relationship to different vegetative plant tissues was investigated. Sugar beet leaves (Beta vulgaris) provided the best active chitinase at optimum extracting conditions (extracted with water, at 16°C for 90 min).
Chitinase from leaves of sugar beet was purified by (NH4)2SO4 precipitation (20-60%) and fractionated on Sephadex G-120 followed by Sephadex G-200 column chromatography. A 18.9 fold purification of the enzyme with 14.0 ImU/mg specific activity was achieved. The yield of the purified chitinase was 43.4 mg protein (608 ImU) from 100g dry leaves tissues. The prepared enzyme was showing a single protein band on agarose gel electrophoresis. The purified enzyme had been shown to have a M, 64×103 Dalton on the basis of gel filtration on Sephadex G-200 column. Optimum chitinase activity on chitin was recorded in 0.1M acetate buffer, pH 4.5 at 40°C.