Identification of a methanol-inducible promoter from Rhodococcus erythropolis PR4 and its use as an expression vector(MICROBIAL PHYSIOLOGY AND BIOTECHNOLOGY)
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概要
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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> 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.
著者
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Tamura Tomohiro
Res. Inst. Of Genome-based Biofactory Aist Toyohira-ku Sapporo 062-8517 Jpn
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Mitani Yasuo
Research Institute Of Genome-based Biofactory National Institute Of Advanced Industrial Science And
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Tamura Tomohiro
Research Institute Of Genome-based Biofactory National Institute Of Advanced Industrial Science And
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Tamura Tomohiro
Bioproduction Research Institute National Institute Of Advanced Industrial Science And Technology (a
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Nakashima Nobutaka
Research Institute Of Genome-based Biofactory National Institute Of Advanced Industrial Science And
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Mitani Yasuo
Bioproduction Research Institute National Institute Of Advanced Industrial Science And Technology (aist)
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Yun Hea-yeon
Bioproduction Research Institute National Institute Of Advanced Industrial Science And Technology (aist):graduate School Of Agriculture Hokkaido University
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Kagawa Yusuke
Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST)
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Nakashima Nobutaka
Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST)
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Tamura Noriko
Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST)
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Nakashima Nobutaka
Bioproduction Research Institute National Institute Of Advanced Industrial Science And Technology (aist)
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Tamura Noriko
Bioproduction Research Institute National Institute Of Advanced Industrial Science And Technology (aist)
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Kagawa Yusuke
Bioproduction Research Institute National Institute Of Advanced Industrial Science And Technology (aist):graduate School Of Agriculture Hokkaido University
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Yun Hea-yeon
Bioproduction Research Institute National Institute Of Advanced Industrial Science And Technology (aist)
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Kagawa Yusuke
Bioproduction Research Institute National Institute Of Advanced Industrial Science And Technology (aist)
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