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Identification of a methanol-inducible promoter from Rhodococcus erythropolis PR4 and its use as an expression vector

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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)

Submitter: 田村 具博

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