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dc.contributor.authorIrla, Marta
dc.contributor.authorHeggset, Tonje Marita Bjerkan
dc.contributor.authorNærdal, Ingemar
dc.contributor.authorPaul, Lidia
dc.contributor.authorHaugen, Tone
dc.contributor.authorLe, Simone Balzer
dc.contributor.authorBrautaset, Trygve
dc.contributor.authorWendisch, Volker F.
dc.identifier.citationFrontiers in Microbiology. 2016, 7 .en_US
dc.description.abstractBacillus methanolicus is a thermophilic methylotroph able to overproduce amino acids from methanol, a substrate not used for human or animal nutrition. Based on our previous RNA-seq analysis a mannitol inducible promoter and a putative mannitol activator gene mtlR were identified. The mannitol inducible promoter was applied for controlled gene expression using fluorescent reporter proteins and a flow cytometry analysis, and improved by changing the -35 promoter region and by co-expression of the mtlR regulator gene. For independent complementary gene expression control, the heterologous xylose-inducible system from B. megaterium was employed and a two-plasmid gene expression system was developed. Four different replicons for expression vectors were compared with respect to their copy number and stability. As an application example, methanol-based production of cadaverine was shown to be improved from 11.3 to 17.5 g/L when a heterologous lysine decarboxylase gene cadA was expressed from a theta-replicating rather than a rolling-circle replicating vector. The current work on inducible promoter systems and compatible theta- or rolling circle-replicating vectors is an important extension of the poorly developed B. methanolicus genetic toolbox, valuable for genetic engineering and further exploration of this bacterium.en_US
dc.publisherFrontiers Mediaen_US
dc.rightsNavngivelse 4.0 Internasjonal*
dc.titleGenome-based genetic tool development for Bacillus methanolicus: Theta-and rolling circle-replicating plasmids for inducible gene expression and application to methanol-based cadaverine productionen_US
dc.typePeer revieweden_US
dc.typeJournal articleen_US
dc.source.journalFrontiers in Microbiologyen_US
dc.description.localcodeopyright © 2016 Irla, Heggeset, Nærdal, Paul, Haugen, Le, Brautaset and Wendisch. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.en_US

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