[[alternative]]Bioconversion of Chitinous Materials by Bacterial Chitinases for the Preparation of Chitooligosaccharides

計畫編號：NSC96-2313-B032-002-MY3研究期間：200908~201007研究經費：1,248,000[[abstract]]本研究擬分三年時間, 以B. amyloliquefaciens V656 所生產幾丁質.為主，配合實 驗室已篩選到四株新穎幾丁質.及/或溶菌.及/或蛋白.生產菌（B. subtilis W-118, B. subtilis TKU007, Bacillus sp. TKU004, Pseudomonas aeruginosa K-187）作為酵素來源。 所擬探討的基質（酵素水解之對象）包括蝦殼、蟹殼、烏賊軟骨、紅麴菌絲以 及其他各種幾丁質及幾丁聚醣產品（不同去乙醯度、分子量）。 『第一年』擬藉由HPLC來探討上述細菌所生產酵素（幾丁質.及/或溶菌.及 /或蛋白.）水解各種基質生產幾丁寡醣（以幾丁六醣及/或七醣為主要目標）之較 適條件，並將所得水解產物進行抗腫瘤測試。 『第二年』擬藉由AS-L這種酸鹼可逆型擔體進行酵素固定化，探討利用固定 化酵素連續生產較多量幾丁寡醣之可行性，並將所得水解產物進行抗腫瘤測試。 『第三年』擬探討這些水解物之免疫活性，以及分析所具抗腫瘤之機轉，探討 所得各種水解產物於抗癌及生醫材料研發之應用潛力。 This is a tree years project. Five bacterial strains , Bacillus amyloliquefaciens V656, B. subtilisW-118, B. subtilis TKU007, Bacillus sp. TKU004, and Pseudomonas aeruginosa K-187 were used for the production of novel chitinases and/or proteases. The chitinous materials used as the enzymatic substrates for preparation of chitooligosaccharides were shrimp shells, squid pen, chitin, chitosan, and cell walls Monascus hyphae. In the first year, the reaction conditions of enzymatic hydrolysis were studied. The produced chitooligosaccharides were analyzed by HPLC. The antitumor activity of these chitooligosaccharides containing hydrolyzates will be analyzed. In the second year, a reversibly soluble polymer (AS-L) will be used as the carrier to immobilized the enzymes produced by above five bacteria strains. The immobilized enzymes will be used to produced large amount of chitooligosaccharides. In the third year, the mechanism of chitooligosaccharides to tumor cells will be study in detail.[[sponsorship]]行政院國家科學委員