Phanaksri Teva, Luxananil Plearnpis, Panyim Sakol, Tirasophon Witoon
Institute of Molecular Biosciences, Mahidol University, Phutthamontol, Nakhon Pathom 73170, Thailand.
Microbial Cell Factory Laboratory, Bioresource Technology Unit, National Center for Genetic Engineering and Biotechnology, Thailand Scinece Park, Pathumthani 12120, Thailand.
J Biosci Bioeng. 2015 Oct;120(4):470-5. doi: 10.1016/j.jbiosc.2015.02.008. Epub 2015 Mar 7.
Strong promoter is an essential factor for production of recombinant protein in various expression systems including Bacillus subtilis. In this study, we described a strategy to improve the expression efficiency using synthetic double promoter. Assembly of the conserved elements from σ(B)- and σ(A)-dependent promoters constitutively improved the yield of recombinant protein approximately 2-3-fold in both exponential and stationary growth phase. The synergistic effect in the double promoter was observed only when σ(B)-promoter was located upstream to σ(A)-promoter but independent to its orientation. A conserved element in either -10 or -35 box of σ(B)-promoter is sufficient to promote the synergism. Hence, this simple strategy of promoter engineering could be an effective way to generate a pool of strong constitutive promoters applicable for heterologous protein expression in B. subtilis in the future.
强启动子是包括枯草芽孢杆菌在内的各种表达系统中重组蛋白生产的关键因素。在本研究中，我们描述了一种使用合成双启动子提高表达效率的策略。来自依赖σ(B)和σ(A)的启动子的保守元件的组装在指数生长期和稳定期均使重组蛋白产量提高了约2至3倍。仅当σ(B)启动子位于σ(A)启动子上游时才观察到双启动子中的协同效应，但其方向无关。σ(B)启动子的-10或-35框中的保守元件足以促进协同作用。因此，这种简单的启动子工程策略可能是未来产生适用于枯草芽孢杆菌中异源蛋白表达的强组成型启动子库的有效方法。
