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Influence of promoter and signal peptide on the expression of pullulanase in Bacillus subtilis |
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| Authors: | Yapin Wang Yihan Liu Zhengxiang Wang Fuping Lu |
| Affiliation: | 1. Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin, 300457, People’s Republic of China 4. The College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, People’s Republic of China 2. National Engineering Laboratory for Industrial Enzymes, Tianjin, 300457, People’s Republic of China 3. Tianjin Key Laboratory of Industrial Microbiology, Tianjin, 300457, People’s Republic of China |
| Abstract: | To achieve efficient expression and secretion of a biologically-active pullulanase, the effect of promoter and signal peptide on the production of pullulanase was studied. Three types of promoters (PP43, P apr and P amy ) and four types of signal peptides (SP sacB , SP amy , SP aprl and SP aprs ) were combined to construct twelve expression cassettes for pullulanase in Bacillus subtilis. The pullulanase activity assay was employed to quantify the level of differential expression, and a real-time PCR assay was applied to comparatively track the transcriptional level. Under the same experimental conditions, the potency ratios among the three promoters were P apr > P amy > PP43. The secretion efficiency ratios mediated by the signal peptides were SP sacB > SP amy > SP aprs > SP aprl . The highest yield of pullulanase could be achieved under the promotion mediated by P apr and secretion by SP sacB . |
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