Isolation and Characterization of Protease Keratinolytic Enzyme in Water Monitor Lizard Intestine.

Abstract

Protease enzyme isolated from mucosa layer in intestine water monitor lizard (Varanus salvator) showed proteolytic activity. Therefore, the purpose of this research was to isolate and characterize of protease produced by water monitor lizard intestine. The intestine mucous layer of water monitor lizard (2%) was dissolved in 50mM buffer phosphate pH 7,0, then centrifuged at 6000 G in 4 ºC temperature for 10 minute. The centrifuged extract was used to analyze the maximum enzyme activity by heating the extract at 45–65 ºC range temperature and 5–20 minute range time. Maximum enzyme activity analysis obtained at 55 ºC for 5 minute for the sample duodenum, jejunum, ileum, colon respectively were 4, 3482 U/ml, 3,9247 U/ml, 3,9176 U/ml, and 3,0353 U/ml. Protease from extract enzyme has seven bands with molecular weight 66,54 kDa, 57,61 kDa, 48,94 kDa, 48,47 kDa, 36,80 kDa, 24,28 kDa, and 12,80 kDa. The enzyme has optimum temperature at 55 ºC and optimum at pH 9, and also inhibited by ion Ca2+ and Mg2+ . The in vitro analysis used chicken powder showed no keratinase activity.