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Identification of a methanol-inducible promoter from Rhodococcus erythropolis PR4 and its use as an expression vector
| Title: | Identification of a methanol-inducible promoter from Rhodococcus erythropolis PR4 and its use as an expression vector |
| Authors: | Kagawa, Yusuke Browse this author | | Mitani, Yasuo Browse this author | | Yun, Hea-Yeon Browse this author | | Nakashima, Nobutaka Browse this author | | Tamura, Noriko Browse this author | | Tamura, Tomohiro Browse this author →KAKEN DB |
| Keywords: | Promoter | | Expression | | Methanol-inducible | | RamB | | Rhodococcus erythropolis |
| Issue Date: | May-2012 |
| Publisher: | Society for Biotechnology, Japan |
| Journal Title: | Journal of Bioscience and Bioengineering |
| Volume: | 113 |
| Issue: | 5 |
| Start Page: | 596 |
| End Page: | 603 |
| Publisher DOI: | 10.1016/j.jbiosc.2011.12.019 |
| Abstract: | The genus Rhodococcus exhibits a broad range of catalytic activity and is tolerant to various kinds of organic solvents. This property makes rhodococci suitable for use as a whole-cell catalyst. Various tools for genetic engineering have been developed to use Rhodococcus erythropolis as a host for bioconversion. In this study, we investigated the protein expression responses of R. erythropolis strains and found that isocitrate lyase production in R. erythropolis PR4 (ICL_[Re]) was induced by methanol. By analyzing the regulation mechanisms of icl_[Re] expression, the ∼200-bp upstream region from the first nucleotide of the translation initiation codon of icl_[Re] was shown to be sufficient for the methanol-inducible expression. Also, the ∼100-bp upstream region exhibited strong constitutive promoter activity by an unknown mechanism(s). By investigating proteins that bound to the upstream region of icl_[Re] in vitro, a RamB homologue of R. erythropolis PR4 (RamB_[Re]) was identified. Moreover, 2 putative RamB_[Re] binding sites were identified in the upstream region of icl_[Re] through pull-down assays. A ramB_[Re] knockout experiment suggested that RamB_[Re] negatively controlled the expression of icl_[Re] and that RamB_[Re] regulation was dependent on the availability of a carbon source. On the basis of these findings, we were able to create novel methanol-inducible and strong constitutive expression vectors. |
| Rights: | © 2012 The Society for Biotechnology, Japan |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/49515 |
| Appears in Collections: | 農学院・農学研究院 (Graduate School of Agriculture / Faculty of Agriculture) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 田村 具博
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