数字农科院2.0

Construction And Application Of A Dual Promoter System For Efficient Protein Production And Metabolic Pathway Enhancement In Bacillus Licheniformis

文献类型： 外文期刊

作者： Rao, Y; Cai, DB; Wang, H; Xu, YX; Xiong, SJ; Gao, L; Xiong, M; Wang, Z; Chen, SW; Ma, X

作者机构：

关键词： Bacillus Licheniformis; Dual-Promoter; Protein Expression; Metabolic Pathway Enhancement

期刊名称： JOURNAL OF BIOTECHNOLOGY

ISSN： 0168-1656

年卷期： 2020 年 312 卷

页码：

收录情况： JCR（2021版） ; EI（2021版）

摘要： Promoter plays the critical role in regulating gene transcription, and dual-promoter has received the widespread attentions due to its high efficiency and continuity, here, we want to construct an efficient dual-promoter for protein production and metabolic pathway enhancement. Firstly, our results indicated that P43 promoter efficiently transcribed at logarithmic period, while the sigma(B)-type promoters (PylB, PgsiB, PykzA) were active at stationary phase. Then, several dual promoters were constructed by coupling these sigma(B)-type promoters with P43, and the attained dual-promoter PykzA-P43 showed the best performance, which led to 1.72-, 3.46- and 1.85-fold increases of green fluorescence intensity, red fluorescence intensity and a-amylase activity, compared with those of the recognized strong promoter P43, respectively. Furthermore, a-amylase activity was further increased to 389.65 U/mL by 32.20 % via optimizing sigma factor binding sites (-10 and -35 boxes) of PykzA-P43, attaining the optimized dual promoter Pdual3. Finally, Pdual3 was applied in metabolic pathway enhancement, and the yields of Poly gamma-glutamic acid, acetoin and 2, 3-butanediol were respectively improved by 82.01 %, 17.09 % and 99.39 %. Our results indicated that dual-promoter significantly enhanced gene expression, and this study provided an energetic dual-promoter Pdual3 for efficient protein production and metabolic pathway enhancement in Bacillus licheniformis.

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