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Cholesterol esterase (CHE; EC 3.1.1.13) hydrolyzes fatty acid esters of cholesterols and is widespread in nature. Mammalian CHEs have been extensively studied, because they have importance in cholesterol absorption and metabolism. Some microbial CHEs are used as diagnostic reagents for measuring total serum cholesterol in blood(1). And also the enzyme
is responsible for the digestion and absorption of lipid nutrients similarly to triglyceride lipase and phospholipase, therefore extensive research has been devoted to the structure-function relationship of this protein as well as its physiological roles in lipid metabolism. Using enzyme
immobilization technology we can extensively utilize the enzyme as a biosensor. A novel CHE genes from Pseudomonas aeruginosa, Vibrio parahaemolyticus, Streptomyces avermitilis are cloned and expressed by E.coli. Using PCR(Polymerase Chain Reaction)technology we cloned
target sequence and also Using PQE30 verctor we transformed it into a competent cell. Protein expression experiment was carried out by IPTG induction and expression level was confirmed by SDS-PAGE. Their cholesterol activity was investigated by Cholesterol/Cholesteryl Ester
Quantitation Kit (BioVision03-100). In further study we will investigate the substrate specificity of the enzyme and immobilization possibility.