幾丁質(zhì)酶的分離純化及其降解性質(zhì)的性質(zhì)研究

生物工程論文編號:SW162  論文字?jǐn)?shù):11543,頁數(shù):27

目錄
中文摘要 I
英文摘要 II
目錄 III
1.緒論 1
 1.1 殼寡糖在醫(yī)藥衛(wèi)生方面的應(yīng)用 2
 1.2 殼寡糖在農(nóng)業(yè)中的應(yīng)用 3
 1.3 殼寡糖在食品工業(yè)的應(yīng)用 3
 1.4 殼寡糖在化妝品方面的應(yīng)用 3
 1.5 殼寡糖的制備方法 3
2.實(shí)驗(yàn)部分 7
 2.1實(shí)驗(yàn)材料和儀器設(shè)備 7
 2.2 幾丁質(zhì)粗酶的制取 9
  2.3 幾丁質(zhì)粗酶的純化 10
 2.4 薄層層析分析酶解產(chǎn)物 11
3.結(jié)果與討論 13
 3.1 濃縮酶液最佳鹽析濃度 13
 3.2 DEAE-Sepharose色譜圖 14
 3.3 Sephacryl S-400色譜圖 15
 3.4 酶純度鑒定和分子量測定 16
 3.5 薄層層析分析酶解產(chǎn)物結(jié)果 17
4.總結(jié)與展望 22
致謝 23
參考文獻(xiàn) 24

 摘要：幾丁質(zhì)是廣泛存在于自然界的一種含氮多糖類生物高分子，其降解產(chǎn)物殼寡糖具有許多優(yōu)異的性能，已被應(yīng)用于生產(chǎn)生活等各方面。幾丁質(zhì)酶是一種降解幾丁質(zhì)的專一性降解酶，降解產(chǎn)物為小分子片段的殼寡糖。本文主要講述從嗜熱芽孢桿菌HD1發(fā)酵產(chǎn)生的粗酶液出發(fā)，通過(NH4)2SO4沉淀、DEAE陰離子交換樹脂層析和Sephacryl S-400凝膠柱層析，得到純化后的幾丁質(zhì)酶，該酶的相對分子質(zhì)量為80.8KD，具有較好的pH穩(wěn)定性和較高的酶活力。經(jīng)過薄層層析對酶解產(chǎn)物的分析表明，此酶對底物幾丁質(zhì)的切割方式為內(nèi)切且降解產(chǎn)物具有良好的水溶解性。
 關(guān)鍵詞：幾丁質(zhì)酶；分離純化；酶解
 
 Abstract: Chitin is widely distributed in nature, a nitrogen-containing polysaccharide biopolymer.Chitosan degradation products have many excellent properties, and have been used in production and other aspects of life.Chitinase is an enzyme which catalyze the glycosidic band of chitin specificitily,and its product is small fragments of chitin.This thesis describes the processes of extracting chitinase produced by HD1 bacteria. The purified enzyme was purified by precipitation,DEAE-Sephadex ion-exchang chromatography and Sephacryl S-400 gel filtration.Its molecular mass was estimated to be about 80.8KD by SDS-PAGE. The hydrolysis product assayed by TLC suggested that the chitinase catalyzed an endo-type cleavage reaction,and the hydrolysis product was water solubility.

Keywords： Chitinase; Purification; Hydrolysis