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DNA Research Advance Access originally published online on March 11, 2008
DNA Research 2008 15(2):73-81; doi:10.1093/dnares/dsn002
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© The Author 2008. Kazusa DNA Research Institute
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org

Enhanced Recombinant Protein Productivity by Genome Reduction in Bacillus subtilis

Takuya Morimoto1,2 {dagger}, Ryosuke Kadoya1,2 {dagger}, Keiji Endo1 {dagger}, Masatoshi Tohata1, Kazuhisa Sawada1, Shengao Liu1, Tadahiro Ozawa1, Takeko Kodama1,3, Hiroshi Kakeshita1,4, Yasushi Kageyama1, Kenji Manabe1, Shigehiko Kanaya2, Katsutoshi Ara1, Katsuya Ozaki1 and Naotake Ogasawara2,*

1 Biological Science Laboratories, Kao Corporation, 2606 Akabane, Ichikai, Haga, Tochigi 321-3497, Japan
2 Graduate School of Information Science, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan
3 Department of Bioscience and Textile Technology, Interdisciplinary Graduate School of Science and Technology, Shinshu University, 3-15-1 Tokida, Ueda, Nagano 386-8567, Japan
4 Graduate School of Life and Environmental Sciences, Tsukuba University, 1-1-1 Ten-noudai, Tsukuba, Ibaraki 305-8572, Japan

Received 24 December 2007 ; accepted 17 January 2008.

The emerging field of synthetic genomics is expected to facilitate the generation of microorganisms with the potential to achieve a sustainable society. One approach towards this goal is the reduction of microbial genomes by rationally designed deletions to create simplified cells with predictable behavior that act as a platform to build in various genetic systems for specific purposes. We report a novel Bacillus subtilis strain, MBG874, depleted of 874 kb (20%) of the genomic sequence. When compared with wild-type cells, the regulatory network of gene expression of the mutant strain is reorganized after entry into the transition state due to the synergistic effect of multiple deletions, and productivity of extracellular cellulase and protease from transformed plasmids harboring the corresponding genes is remarkably enhanced. To our knowledge, this is the first report demonstrating that genome reduction actually contributes to the creation of bacterial cells with a practical application in industry. Further systematic analysis of changes in the transcriptional regulatory network of MGB874 cells in relation to protein productivity should facilitate the generation of improved B. subtilis cells as hosts of industrial protein production.

Key words: Bacillus subtilis; genome size reduction; recombinant protein productivity

* To whom correspondence should be addressed. Tel. + 81 743-72-5430. Fax. + 81 743-72-5439. E-mail: nogasawa{at}bs.naist.jp

Edited by Katsumi Isono

{dagger} These authors contributed equally to this work.


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