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題 名 | Effects of Culturing Conditions on Production of D-hydantoinase from Recombinant Escherichia Coli=利用重組大腸桿菌生產D-Hydantoinase |
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作 者 | 黃健雄; 許文輝; 陳俊瑋; | 書刊名 | 微免與感染雜誌 |
卷 期 | 37:6 2004.12[民93.12] |
頁 次 | 頁313-321 |
分類號 | 414.83 |
關鍵詞 | 重組; 大腸桿菌; Escherichia coli; Hydantoinase; Inclusion bodies; Plasmids; Recombinant proteins; |
語 文 | 英文(English) |
英文摘要 | The effects of culturing conditions on D-hydantoinase production by a recombinant Escherichia coli strain were investigated using a controlled fed-batch fermentation system. Glucose concentration and pH of the culture broth were maintained at less than 3.3 g/L and at 7.0, respectively, in a 5 L jar fermentor. The optimal composition of the batch medium was glucose, 0.25%; yeast extract, 0.75%; (NH4)2SO4, 0.25%; KH2PO4, 0.2%. The optimal feeding solution was glucose, 60%; yeast extract, 30%; ammonia water, 9%. Following 25-h cultivation, 0.02 mM isopropyl-s-thiogalactopyranoside was added to induce dht gene expression and the temperature was shifted from 32 to 27° C to avoid inclusion body formation. The plasmid-harboring dht gene was found to be stably maintained in the E. coli. Under optimal conditions, a cell density of about 25 g dry cell weight/L and a high volumetric productivity of 8300 U/L/h could be achieved after 48 h culture at agitation and aeration rates of 1000 revolutions per minute and 1 vvm, respectively. |
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