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題 名 | 篩選枯草菌之DNA轉形株用於蛋白酶之生產 |
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作 者 | 王美人; 游若篍; | 書刊名 | 食品科學 |
卷 期 | 22:1 1995.02[民84.02] |
頁 次 | 頁38-45 |
分類號 | 463.824 |
關鍵詞 | DNA轉形株; 枯草桿菌; 蛋白酶生產; 營養需求株; DNA transformants; Bacillus subtilis; Protease production; Auxotrophs; |
語 文 | 中文(Chinese) |
中文摘要 | 本試驗以Bacillus subtilis DB403,作為DNA轉形之受體株。DB403首先利用亞硝基胍處理,以篩選營養突變株。以B. subtilis CCRC11199,一株蛋白?生產菌株,抽取之DNA為材料,並以並同之營養需求當作標誌進行DNA轉形,自篩得高蛋白?基因特性之轉形株,以三角瓶進行蛋白?生產,結果分別選出中性及鹼性生產,結果分別選出中性及鹼性蛋白?高產株L3及M10。當以四種不同米粉顆粒之LB培養基培養時,發現L3在1%蓬來胚芽米粉顆粒之LB培養基,於37°C培養24小時,生產最高量之中性蛋白?(194 U/ml);M10則在1%在來米粉顆粒之LB培養基,於37°C培養48小時,生產最高量之鹼性蛋白?(166 U/ml)。進一步以3%米粉顆粒之LB培養基,培養轉形株L3和M10時,其蛋白?之產量並不理想,因此欲利用米為基質生產蛋白?,尚需作進一步的探討。 |
英文摘要 | Bacillus subtilis tranformants which secrete high levels of protease were constructed and screened. Firstly, B subtilis DB403, a protease-deficiency strain, was treated with nitrosoguanidine and hundreds auxotrophs were isolated from it. Then, chromosome DNA was isolated from B. subtilis CCRC11199, a protease producer, and was transferred to auxotrophs of B. subtilis DB403 to isolate protease producer. Based on the protease production experiment, L3 and M10transformants were separately considered as the highest producer of neutral and alkaline protease. Being cultivated in four different kinds of rice powders in LB media, L3 produced the highest amount of neutral protease on a LB medium plus 1% Japonica germ rice powders after 24 hours of incubation at 37°C; while M10 produced the highest amount of alkaline protease on a LB medium plus 1% Indica polished rice powders after 48 hours of incubation at 37°C. However, the enzyme activity was not increased significantly when these substrate concentrations were increased from 1 to 3%. Therefore, further experiments are highly necessary for a scale-up production of protease. |
本系統中英文摘要資訊取自各篇刊載內容。