| 运用试验设计方法提高胞外杂合b-葡聚糖酶在重组大肠杆菌中的表达 |
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| 引用本文： | 卢英华,邓旭,程志敬,李清彪,刘刚.运用试验设计方法提高胞外杂合b-葡聚糖酶在重组大肠杆菌中的表达[J].中国化学工程学报,2007,15(2):172-177. |
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| 作者姓名： | 卢英华 邓旭 程志敬 李清彪 刘刚 |
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| 作者单位： | [1]Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China [2]College of Life Science, Shenzhen University, Shenzhen 518060, China [3]The Key Laboratory for Chemical Biology of Fujian Province, Xiamen University, Xiamen 361005, China |
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| 基金项目： | Supported by the National Natural Science Foundation of China （No.20306025） and the Xiamen Science and Technology Project （No.3502Z20055017）. |
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| 摘 要： | A genetically engineered Escherichia coli JM109 harboring pLF3 was used to produce a hybrid extracellular β-glucanase. Starting with enzyme production medium, glycerol and yeast extract combined with NaNO3 were screened to be the most suitable carbon and nitrogen source, respectively. Analysis of six components of the enzyme production medium by employing statistical optimization methods such as Plackett-Burman design and steepest ascent showed that yeast extract was the only significant variable and its best concentration for enzyme production was 12g·L^-1. After optimization of the medium, 297.71U·ml^-1 of β-glucanase activity in the medium and 352350U·g^-1 of β-glucanase selectivity could be obtained, which were 14 and 72 folds higher than those obtained from original medium, respectively. Even higher enzyme activities were achieved by batch cultivations in a conventional stirred bioreactor on the optimized medium.
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| 关 键 词： | 胞外β-葡聚糖酶 强化生产 重组大肠杆菌 实验设计方法 |
| 收稿时间： | 31 August 2006 |
| 修稿时间： | 2006-08-312007-03-05 |
Enhanced Production of Hybrid Extracellular β-Glucanase by Recombinant Escherichia coli Using Experimental Design Method |
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| LUYinghua,DENGXu,CHENGZhijing,LIQingbiao,LIUGang.Enhanced Production of Hybrid Extracellular β-Glucanase by Recombinant Escherichia coli Using Experimental Design Method[J].Chinese Journal of Chemical Engineering,2007,15(2):172-177. |
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| Authors: | LUYinghua DENGXu CHENGZhijing LIQingbiao LIUGang |
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| Affiliation: | 1. Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China;2. College of Life Science, Shenzhen University, Shenzhen 518060, China;3.The Key Laboratory for Chemical Biology of Fujian Province, Xiamen University, Xiamen 361005, China |
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| Abstract: | A genetically engineered Escherichia coli JM109 harboring pLF3 was used to produce a hybrid extracellular β-glucanase. Starting with enzyme production medium, glycerol and yeast extract combined with NaNO3 were screened to be the most suitable carbon and nitrogen source, respectively. Analysis of six components of the enzyme production medium by employing statistical optimization methods such as Plackett-Burman design and steepest ascent showed that yeast extract was the only significant variable and its best concentration for enzyme production was 12g.L-1. After optimization of the medium, 297.71U.ml-1 of β-glucanase activity in the medium and 352350U.g-1 of β-glucanase selectivity could be obtained, which were 14 and 72 folds higher than those obtained from original medium, respectively. Even higher enzyme activities were achieved by batch cultivations in a conventional stirred bioreactor on the optimized medium. |
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| Keywords: | β -glucanase recombinant Escherichia coli statistical methodology medium optimization |
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