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頁籤選單縮合
題名 | Production of Heterologous Providencia rettgeri Penicillin Acylase in Escherichia Coli=於大腸菌中進行Providencia rettgeri Penicillin Acylase生產之異型基因表現 |
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作者姓名(中文) | 周志雄; 林明毅; 王文昌; | 書刊名 | Journal of the Chinese Institute of Chemical Engineers |
卷期 | 31:2 2000.03[民89.03] |
頁次 | 頁135-144 |
分類號 | 364.21 |
關鍵詞 | 大腸菌; 異型基因表現; Heterologous expression; Native expression; Penicillin acylase; Cloning; Recombinant DNA technology; Catabolite repression; |
語文 | 英文(English) |
中文摘要 | 本研究為植株Prouidencia rettgeri 之 pac基因,並於大腸桿菌中進行異型基因表現,並探討各種環境因素對其基因表現之影響。 |
英文摘要 | In this work, we constructed several expression plasmids for the production of Pravidencia rettgeri penicillin acylase (EC 3.5.1. 11; PAC) in Escherichia coli. DNA fragments containing the pac gene from P. rettgeri ATCC31O52 were PCR-amplified and cloned in-to the expression vectors so that the pac gene expression was controlled by the lac or trc pro-mater system. The effects of culture conditions, such as IPTG concentration, temperature, and carbon source, on the native or heterologous expression were investigated. Among a selection of expression systems, JM1O9 harboring pUTKnPAC2601 gave the highest PAC activity and could be of interest for industrial application. Cultivation should be perfonned at a temperature ranging from 28℃ to 33℃ and the medium could be supplemented with glycerol. The host/vector sys-tern offers an opportunity for high-temperature-oriented PAC production, which is usually con-ducted at a low temperature. Volumetric PAC activity at more than fiftyfold (~820 U/L) that of the native expression in ArCC31O52 (~15 U/L) could be reached by optimization of the host/vector system and culture conditions. |
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