Enhanced Extracellular Production of Heterologous Proteins in Bacillus subtilis by Deleting the C terminal Region of the SecA Secretory Machinery英文文献.pdfVIP

Mol Biotechnol (2010) 46:250–257 DOI 10.1007/s12033-010-9295-0 RESEARCH Enhanced Extracellular Production of Heterologous Proteins in Bacillus subtilis by Deleting the C-terminal Region of the SecA Secretory Machinery Hiroshi Kakeshtia • Yasushi Kageyama • Katsutoshi Ara • Katsuya Ozaki • Kouji Nakamura Published online: 24 June 2010 Springer Science+Business Media, LLC 2010 Abstract In this study, we examined the effects of modi- Introduction fying the C-terminal region of the SecA protein on the pro- duction of heterologous proteins in Bacillus subtilis . SecA Bacillus subtilis and related species are widely used as was selected as a candidate among the components of the Sec hosts for the extracellular production of industrially worthy system due to its ability to interact directly with both the enzymes, such as amylases, proteases, and lipases [ 1–3]. precursors and membrane translocases. A phylogenetic These species possess a very high capacity for secreting a comparison demonstrated that the C-terminal region is not variety of exoenzymes into the growth medium, thereby well conserved among eubacterial SecA proteins. The reducing downstream purification processes. Therefore, deletion of the 61 amino acids at the C-terminal region led to developing enzyme-producing hosts is a major challenge an 83% increase in extracellular alkaliphilic Bacillus sp. in the field of biotechnology and protein production. In thermostable alkaline cellulase (Egl-237) activity. More- B. subtilis, most secreted proteins are translocated across over, the productivity of human interferon a (hIFN-a2b) was the cytoplasmic membrane via the Sec system [2, 4, 5]. increased by 2.2-fold compared to the wild-type SecA, by Almost