Enhanced production of hybrid extracellular β-glucanase by recombinant Escherichia coli  using experimental design method

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Enhanced production of hybrid extracellular β-glucanase by recombinant Escherichia coli
using experimental design method

LUYinghua^a,c; DENGXu^b;CHENGZhijing^a;LIQingbiao^a,c;LIUGang^b

^a Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China

^b College of Life Science, Shenzhen University, Shenzhen 518060, China
^c The Key Laboratory for Chemical Biology of Fujian Province, Xiamen University, Xiamen 361005, China

Received:1900-01-01 Revised:1900-01-01 Online:2007-04-28 Published:2007-04-28
Contact: LU Yinghua

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

卢英华^a,c; 邓旭^b; 程志敬^a; 李清彪^a,c; 刘刚^b

^a Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China ^b College of Life Science, Shenzhen University, Shenzhen 518060, China
^c The Key Laboratory for Chemical Biology of Fujian Province, Xiamen University, Xiamen 361005, China

通讯作者: 卢英华

Abstract

Abstract: A genetically engineered Escherichia coli JM109 harboring pLF3 was used to produce a hybrid extracellular -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 ob-tained from original medium, respectively. Even higher enzyme activities were achieved by batch cultivations in a conventional stirred bioreactor on the optimized medium.

Key words: β-glucanase, recombinant Escherichia coli, statistical methodology, medium optimization

摘要： A genetically engineered Escherichia coli JM109 harboring pLF3 was used to produce a hybrid extracellular -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 ob-tained from original medium, respectively. Even higher enzyme activities were achieved by batch cultivations in a conventional stirred bioreactor on the optimized medium.

关键词: β-glucanase;recombinant Escherichia coli;statistical methodology;medium optimization

LUYinghua, DENGXu,CHENGZhijing,LIQingbiao,LIUGang. Enhanced production of hybrid extracellular β-glucanase by recombinant Escherichia coli
using experimental design method[J]. .

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]. CIESC Journal.

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