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題 名 | Listonella damsela NTU-FC-6幾丁質酶之生產與基本性質之探討=Production and Some Properties of Chitinase (s) from Listonella Damsela NTU-FC-6 |
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作 者 | 蘇南維; 李敏雄; | 書刊名 | 中國農業化學會誌 |
卷 期 | 36:1 1998.02[民87.02] |
頁 次 | 頁65-76 |
分類號 | 346.217 |
關鍵詞 | 幾丁質; 幾丁質; Chitin; Chitinase; Listonella damsela; |
語 文 | 中文(Chinese) |
中文摘要 | 以膠態幾丁質為積聚培養的主要碳源,本研究成功地分離出對幾丁質具有高分解 能力之菌株 Listonella damsela NTU-FC-6, 於最適培養條件下( colloidal chitin 1.5 %, malt extract 1.5 %,K �� HPO �� 0.05 %, KH �� PO �� 0.1 %,MgSO �瓷D 7H �� O 0.05 %,pH 7.0,30 ℃ 110 rpm )振盪培養 72 小時,發酵液之幾丁質�t活性可達 1.8 units ╱ mL。 Listonella damsela NTU-FC-6 所產生之幾丁質�t經初步純化,發酵液 以超過濾裝置濃縮 10 倍體積, 收集 30 ﹣ 90 %( v ╱ v )丙酮沉澱區間, 再以 4 M 氯化鈉水溶液溶解,去除沉澱部份後,幾丁質分解酵素活性回收 45 %,比活性 0.35 unit ╱ mg。 經氯化鈉區分之酵素液對幾丁質的基質特異性良好,對幾丁聚醣只具輕微的水解能 力; 經氯化鈉區分之酵素液再由 Sephacryl S-200 膠體層析進一步的純化可得兩個幾丁質 �t活性峰,分別為幾丁質�t A 及幾丁質�t B; 將此二幾丁質�t再經製備式等電焦集電泳分 析結果,幾丁質�t A 出現單一活性峰,為幾丁質�tⅠ;幾丁質�t B 有兩個酵素活性峰,分 別為幾丁質�tⅡ及幾丁質�tⅢ。幾丁質�t A 與幾丁質�t B 分別與膠態幾丁質作用之產物組 成與經氯化鈉區分之酵素之反應結果類似,產物組成皆以 N,N' ﹣二乙醯幾丁二糖為主,佔 80 %以上,另含少量的 N ﹣乙醯葡萄糖胺與其三聚醣。 |
英文摘要 | A bacterial strain secreting potent chitinolytic activity was isolated from soil by means of an enrichment culture using colloidal chitin as the major carbon source. The isolated bacterium, designated as Listonella damsela NTU-FC-6, under optimal culture conditions, produced a chitinolytic activity of 1.8 units/mL after 72 hours of incubation in a 500 mL shaker flask containing 100 mL medium. The secretion of the enzyme was stimulated by several organic nitrogenous and fewer carbonous supplements. Colloidal chitin (1.5%) and malt extract (1.5%) displayed better concentractions for enzyme secretion. The production of chitinolytic enzyme was enhanced by increasing the concentration of the malt extract to 1.5%. The chitinase(s) was purified by concentrating the supernatant medium to 1/10 volume and fractionating using acetone solvent. The sediments were dissolved using 4 M NaCl(aq), and then impurities were removed through centrifugation, followed by Sephacryl S-200 gel permeation chromatography. Two major peaks found in the chromatogram were named chitinase A and chitinase B. The chitinase A and chitinase B were further purified by means of preparative isoelectric focusing electrophoresis. One chromatogram showed one peak from chitinase A, named chitinase Ⅰ, and the other chromatogram contained two major peaks from chitinase B, named chitinase Ⅱ and chitinase Ⅲ. Chitinase A and chitinase B hydrolyzed colloidal chitin, and the products of the reaction as same as the enzyme from NaCl fractionatation were mainly composed of N,N'-diacetylchitobiose with smaller amounts of N-acetylglucosamine and trimer. |
本系統中英文摘要資訊取自各篇刊載內容。