胰蛋白酶的固定化及其水解酪蛋白的研究

生物工程論文編號:SW131  論文字數:10553,頁數:23

目 錄
中文摘要
英文摘要…………………………………………………………………….…..…Ⅱ
目錄…………………………………………………………………………..…….Ⅲ
緒論………………………………………………………………………..……1
 1.1 胰蛋白酶的研究概述……………………………………………….……1
 1.1.3 胰蛋白酶的制備方法………………………………………………2
 1.2 固定化…………………………………………………………………….4
 1.3 胰蛋白酶的固定化研究現狀………………………………………….…6
 1.4  本文的研究思路和目標……………………………………………...…8
2. 實驗部分……………………………………………………………………...…9
 2.1 用樹脂作介質固定化…...……………………………………………..…9
 2.2 以海藻酸鈣為載體固定化胰蛋白酶…………………………………..14
3. 結果與討論………………………………………….……………………..….18
 3.1實驗結果………………………………………...……………...……….18
 3.2實驗討論……………………………………………………...…………18
4. 總結與展望……………………….………………………………………...…19
致謝………………………………….…………………………………………….20
參考文獻……………………………………….………………………………….21

摘要：胰蛋白酶是一種動物來源的蛋白水解酶，分子量為23.8 kD，在醫藥、食品、工業等領域有廣泛的應用。本文研究了胰蛋白酶的固定化方法以及利用固定化胰蛋白水解酪蛋白的過程和相關工藝。論文在建立測定胰蛋白酶活力方法的基礎上，采用吸附法篩選出適合固定化胰蛋白酶的大孔弱陰離子交換樹脂Duolite A7，優化的樹脂添加量為0.5g/10mg 蛋白，固定化催化5批次反應后仍有30％以上的酶活力； pH、溫度、包埋量等條件對海藻酸鈣固定化胰蛋白酶活力的影響進行了考察，海藻酸鈉的濃度為15 g/L，CaCl2的濃度為20 g/L，胰蛋白酶的包埋量為60mg/10mL海藻酸鈉，反應最適的溫度和pH分別為60℃和7.4。
關鍵詞：胰蛋白酶；固定化；海藻酸鈣；Duolite A7樹脂
 

Abstract: The trypsin that the molecular weight is 23.8 kD is the hydrolytic enzyme which comes from animal. It has the extensive application in medicine, food, industry and so on. This article has studied the trypsin immobilization method as well as the use of trypsin immobilization for casein hydrolysis. Based on determination of trypsin enzyme activity, a macropore weak anion exchange resin Duolite A7 was screened for trypsin immobilization, the optimized resin addition amount is 0.5g/10mg protein, and enzyme activity still reached more than 30% after five-time recycling of the immobilized enzyme. Conditions on pH, temperature, entrapment amount on effect of trypsin activity were investigated by Ca-alginate gel beads, the sodium alginate concentration is 15 g/L, CaCl2 20 g/L, the trypsin entrapment amount is 60mg/10mL sodium alginate; the optimal temperature and pH are 60℃ and 7.4 respectively.
Keywords: Trypsin; Immobilizition; Ca-alginate; Duolite A7 resin