Liu Xin, Wang Hai, Wang Bin, Pan Li
School of Biology and Biological Engineering, South China University of Technology, Guangzhou, 510006, China.
Protein Expr Purif. 2018 Nov;151:72-77. doi: 10.1016/j.pep.2018.06.006. Epub 2018 Jun 9.
Bacillus subtilis is widely used for the large-scale industrial production of proteins. In this study, the transcriptomes of B. subtilis 168 and B. megaterium DSM319 cells grown in stationary phase were analyzed to expand the repertoire of highly-active promoters for high-level protein expression based on the transcriptomes of these Bacillus strains. 24 genes with the highest expression levels among 2048 highly expressed gene families were chosen to examine promoter activity. The activities of four promoters with the beta-galactosidase (bgaB) gene as a reporter were stronger than those of the well-characterized strong promoter P43. The expression level of recombinant Pro-transglutaminase (pro-MTG) from Streptomyces mobaraensis achieved 87.6 U/mL and 70.7 U/mL under the control of two constitutive promoter P and P, respectively, compared to the promoter P43. Our study provides a basis for further studies on the Bacillus transcriptome by identifying strong promoters for industrial uses.
枯草芽孢杆菌被广泛用于蛋白质的大规模工业生产。在本研究中，对处于稳定期生长的枯草芽孢杆菌168和巨大芽孢杆菌DSM319细胞的转录组进行了分析，以基于这些芽孢杆菌菌株的转录组扩展用于高水平蛋白质表达的高活性启动子库。从2048个高表达基因家族中选择了24个表达水平最高的基因来检测启动子活性。以β-半乳糖苷酶（bgaB）基因为报告基因的四个启动子的活性比已充分表征的强启动子P43更强。与启动子P43相比，来自茂原链霉菌的重组谷氨酰胺转胺酶（pro-MTG）在两个组成型启动子P和P的控制下，表达水平分别达到87.6 U/mL和70.7 U/mL。我们的研究通过鉴定用于工业用途的强启动子，为进一步研究芽孢杆菌转录组提供了基础。
