Expression of Bacillus acidopullulyticus pullulanase in Escherichia coli and its optimization of fermentation

Authors

YANG Xianghui, State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China; School of Biotechnology, Key Laboratory of Industrial Biotechnology, by the Ministry of Education, Jiangnan University, Wuxi, Jiangsu 214122, China; Joi
CHEN Sheng, State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China; School of Biotechnology, Key Laboratory of Industrial Biotechnology, by the Ministry of Education, Jiangnan University, Wuxi, Jiangsu 214122, China; Joi
WU Jing, State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China; School of Biotechnology, Key Laboratory of Industrial Biotechnology, by the Ministry of Education, Jiangnan University, Wuxi, Jiangsu 214122, China; Joi

Abstract

To increase expression and extracellular secretion of pullulanase of the recombinant strain E.coli BL21(DE3)/pET20b(+)-BapulA at 3 L fermentor level. Expression of recombinant pullulanase in E. coli and further optimization of fermentation at the 3 L fermenter Firstly. The effects were investigaged, including the fermentation temperature, pH, IPTG concentration, induction time and glycine concentration on the expression of recombinant pullulanase. The optimum fermentation conditions were showed as followed: the growth temperature and pH of the pre-induction cells were 30 ℃ and 7.0, respectively. When the cell density reached 50 OD₆₀₀, the fermentation temperature was shifted to 25 ℃, and the pH was adjusted to 6.2 with lactose flow rates of 0.4 g/(L·h). During the fermentation process, glycine was added at the concentration of 1.5 g/L when the cell density of OD₆₀₀ reached 15, 45 and 75, respectively. When OD₆₀₀ reached 105, glycine was added at a concentration of 3 g/L. The optimal total and extracellular pullulanase activity were 1 910.1 U/mL and 659.0 U/mL, respectively, which represent 45.4 and 100-fold compared with those observed under initial conditions.

Publication Date

5-28-2018

First Page

20

Last Page

26

DOI

10.13652/j.issn.1003-5788.2018.05.005

Recommended Citation

Xianghui, YANG; Sheng, CHEN; and Jing, WU (2018) "Expression of Bacillus acidopullulyticus pullulanase in Escherichia coli and its optimization of fermentation," Food and Machinery: Vol. 34: Iss. 5, Article 5.
DOI: 10.13652/j.issn.1003-5788.2018.05.005
Available at: https://www.ifoodmm.cn/journal/vol34/iss5/5

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