利用海藻酸鈉凝膠固定化Aeromonas caviae NCHU1菌體生產幾丁酵素

Production of Chitinase by Immobilized Aeromonas caviae NCHU1 Coagulated with Sodium Alginate

呂昇儒(Sheng-Ru Lu)

指導教授 ： 張永吉

中臺科技大學/健康科學院/食品科技研究所/碩士(2011年)

摘要

利用固定化菌體生產幾丁酵素，具有連續大量生產之潛力。本研究採用台灣中油公司煉製研究所篩選，並經食品工業發展研究所鑑定之菌株Aeromonas caviae NCHU1，以海藻酸鈉凝膠技術將菌體固定化後，探討其生產幾丁酵素之可行性。菌體固定化探討條件包括:海藻酸鈉濃度(2.0~3.0 %)、乳酸鈣溶液濃度(2.0~4.0 %)及鈣離子溶液浸漬時間(20~40 minutes)等。此外，將最適固定化A. caviae NCHU1菌體置於含幾丁質液體培養基溶液中進行震盪培養，分析其液體培養發酵液中，還原糖濃度與幾丁酵素之活性。試驗結果顯示，最適菌體固定化之條件

將菌體固定於海藻酸鈉-乳酸鈣的凝膠顆粒中，於幾丁質液態培養液中震盪培養48 hr(28 ℃、200 rpm)。期間取固定化菌體培養液，分別檢測還原糖濃度、幾丁酵素活性與菌數，並與未固定化菌體培養作比較。結果顯示，
液體培養液中，還原糖濃度與酵素活性分別於培養12 hr（0.049 mg/ml）及24 hr達到最高值(0.531 U/ml)，前者分析值低於未固定化培養（0.074 mg/ml），後者分析值則略高於未固定化培養(0.462 U/ml)之對照組。雖然有部份菌體外移至固定化膠體外的培養液中，但本研究固定化菌體膠體技術，仍可以提供幾丁質酵素連續生產作業之良好模式。
(1) 酵素活性之比較發現固定化酵素活性於24hr時產量(0.531 U/ml)略高於未固定化12hr(0.462 U/ml)，另外發現由於固定化之酵素活性達最高點時間比未定化晚了12小時(圖4-10）。

關鍵字

Aeromonas caviae ； 幾丁質 ； 液態培養 ； 幾丁酵素 ； 固定化菌體

並列摘要

Abstract
The objective of this study was to investigate the production of chitinase by the immobilized cells of Aeromonas caviae NCHU1 coagulated with sodium alginate. The optimization of the cells immobilized with the sodium alginate-calcium system was studied by evaluation of the free cells’ number leaks to broth culture. The cells’ immobilized conditions were investigated including the concentration of sodium alginate(4.0~6.0 %), calcium lactate(2.0~3.0 %) and the soaking time (20~40 minutes) of the coagulated beads in the calcium lactate solution. The broth cultures(chitin contained) of coagulated beads of the A. caviae NCHU1 for the production of chitinase were evaluated by analysis of enzyme activity and were compared with that of non-immobilized cells. The results showed that the optimized condition for the immobilization were performed with sodium alginate 2.5 % , calcium lactate 3.0 % and the soaking time 30 minutes for the coagulated beads in the calcium lactate solution. The chitinase activities of the immobilized A. caviae NCHU1 was 0.531 U/ml at cultured time 24 hr higher than that of non-immobilized cells 0.462 U/ml. The results showed the potential of the chitinase production by the chitin-broth culture of immobilized A. caviae NCHU1 cells coagulated with sodium alginate-Calcium system. Further studies for the broth culture with continue process for commercial application are necessary and the above results might be applied for the advanced investigation.

並列關鍵字

cells’ immobilization ； chitinase ； chitin ； brth culture ； Aeromonas caviae

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