四种选定蛋白及其水解物中降胆固醇作用之研究.doc

四種選定蛋白及其水解物中降膽固醇作用之研究 江文德＊　劉宜洵　盧錫祺 東海大學食品科學系 （收件日期：95年5月11日；接受日期：95年7月5日） 摘　　要 　　本研究運用CaCo-2細胞模擬小腸上皮細胞，以探討蛋白質及其水解物對抑制細胞膽固醇吸收量的影響，若CaCo-2細胞對膽固醇的吸收量愈低，表示受測物的降膽固醇作用愈強。首先比較四種蛋白質原料包括分離大豆蛋白（ISP）、小麥蛋白、乳清蛋白與卵蛋白對CaCo-2細胞膽固醇吸收的影響，其中以ISP的降膽固醇作用最強。進一步將 ISP分別以三種腸內酵素進行水解，包括胰蛋白酉每、胰凝乳酉每與胃蛋白酉每等，以探討ISP在酵素水解前後對CaCo-2細胞相對膽固醇吸收率（RCU, %）的影響，結果顯示經酵素水解所得之ISP水解物與未水解的 ISP比較均可顯著（p 0.05）降低其CaCo-2細胞的RCU至20%以下。當ISP分別以此三種酵素水解並控制適當的水解率（DH, %）時，每一酵素均可獲得具有最低膽固醇吸收量的水解物，其中以胃蛋白酉每水解ISP的DH控制在12.1%時所得之水解物，其膽固醇吸收量最低可達1.61 pmol/well。其次，依據胃蛋白酉每水解 ISP期間所得水解物之分子量分布與其DH的關係中發現，水解物中 10,000 daltons的胜月太，為抑制CaCo-2細胞對膽固醇吸收的主要成份。 關鍵詞：分離大豆蛋白、蛋白水解物、CaCo-2細胞、降膽固醇作用 Studies of Cholesterol-lowering Actions of Four Given Proteins and Their Hydrolysates Wen-Dee Chiang＊, I-Hsun Liu and Hsi-Chi Lu Graduate Institute of Food Science, Tunghai University, Taichung, （Date Received: May 11, 2006; Date Accept: July 5, 2006） Abstract The measurement on the amount of cholesterol uptake in CaCo-2 cells was employed to evaluate the ability of proteins and their hydrolysates to inhibit cholesterol uptake of cells. The lower the cholesterol uptake represents the higher the cholesterol-lowering action. Among four given proteins, the isolated soy protein (ISP) showed the lowest cholesterol uptake in CaCo-2 cells. Furthermore, the enzymatically hydrolyzed ISP was studied for their effect on the cholesterol-lowering action. Three different proteolytic enzymes including trypsin, chymotrypsin and pepsin were used to hydrolyze ISP. Each reaction solution was centrifuged to obtain supernatant to produce the hydrolysate. The relative cholesterol uptake (RCU, %) was determined by comparing the cholesterol-lowering effects of the hydrolysates against that of intact ISP. The results showed that the RCU values of all hydrolysates were significantly (p0.05) lower than that of intact ISP. By controlling optimum degree of hydrolysis (DH, %), the products of three enzymes all can significantly lower the cholesterol uptake in CaCa-2 cells. Never