Protein Science Group, Department of Biosciences, University of Kent, Canterbury, Kent, CT27NJ, UK.
Adv Biochem Eng Biotechnol. 2010;120:133-61. doi: 10.1007/10_2009_62.
Over the last two decades the Gram-positive bacterium Bacillus megaterium was systematically developed to a useful alternative protein production host. Multiple vector systems for high yield intra- and extracellular protein production were constructed. Strong inducible promoters were combined with DNA sequences for optimised ribosome binding sites, various leader peptides for protein export and N- as well as C-terminal affinity tags for affinity chromatographic purification of the desired protein. High cell density cultivation and recombinant protein production were successfully tested. For further system biology based control and optimisation of the production process the genomes of two B. megaterium strains were completely elucidated, DNA arrays designed, proteome, fluxome and metabolome analyses performed and all data integrated using the bioinformatics platform MEGABAC. Now, solid theoretical and experimental bases for primary modeling attempts of the production process are available.
在过去的二十年中，革兰氏阳性菌巨大芽孢杆菌被系统地开发为一种有用的蛋白质生产替代宿主。构建了多个用于高产细胞内和细胞外蛋白质生产的载体系统。强诱导启动子与优化核糖体结合位点、各种用于蛋白质输出的引导肽、N 和 C 末端亲和标签的 DNA 序列相结合，用于所需蛋白质的亲和色谱纯化。已成功测试了高密度细胞培养和重组蛋白生产。为了进一步基于系统生物学控制和优化生产过程，对两种巨大芽孢杆菌菌株的基因组进行了完全阐明，设计了 DNA 芯片，进行了蛋白质组、通量组和代谢组分析，并使用生物信息学平台 MEGABAC 整合了所有数据。现在，生产过程的初步建模尝试有了坚实的理论和实验基础。
