Production of a Thermostable Pullulanase in Bacillus subtilis by Optimization of the Expression Elements
Bo Pang
The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122 China
Search for more papers by this authorLi Zhou
The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122 China
Search for more papers by this authorWenjing Cui
The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122 China
Search for more papers by this authorZhongmei Liu
The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122 China
Search for more papers by this authorCorresponding Author
Zhemin Zhou
The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122 China
E-mail: [email protected]
Search for more papers by this authorBo Pang
The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122 China
Search for more papers by this authorLi Zhou
The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122 China
Search for more papers by this authorWenjing Cui
The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122 China
Search for more papers by this authorZhongmei Liu
The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122 China
Search for more papers by this authorCorresponding Author
Zhemin Zhou
The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122 China
E-mail: [email protected]
Search for more papers by this authorAbstract
Pullulanase is a debranching enzyme commonly used in the starch processing industry. Production of a thermostable pullulanase with high activity is desired due to the application of the enzyme to starch processing at high temperature. In this study, a thermostable pullulanase with high activity is overproduced in Bacillus subtilis. Via improvement of the plasmid backbones, promoters, and ribosome-binding-site (RBS) sequences, an optimal recombinant strain B. subtilis WB800/RBS7 is obtained with an extracellular pullulanase activity of 154.9 U mL−1 in a shake flask, which is 136.8 times higher than that of the wild type. When the strain is cultured in a 5-L bioreactor, the pullulanase activity is significantly improved to 269.1 U mL−1 (2.74 mg mL−1). These results are expected to aid pullulanase production in industrial starch debranching applications.
Conflict of Interest
The authors declare no conflict of interest.
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