1887

Abstract

strain UT26, whose γ-hexachlorocyclohexane-degrading ability has been studied in detail, is a typical aerobic and heterotrophic bacterium that needs organic carbon sources for its growth, and cannot grow on a minimal salt agar medium prepared without adding any organic carbon sources. Here, we isolated a mutant of UT26 with the ability to grow to visible state on such an oligotrophic medium from a transposon-induced mutant library. This high-yield growth under oligotrophic conditions (HYGO) phenotype was CO-dependent and accompanied with CO incorporation. In the HYGO mutant, a transposon was inserted just upstream of the putative Zn-dependent alcohol dehydrogenase (ADH) gene () so that the gene was constitutively expressed, probably by the transposon-derived promoter. The -deletion mutant (UT26DAX) harbouring a plasmid carrying the gene under the control of a constitutive promoter exhibited the HYGO phenotype. Moreover, the HYGO mutants spontaneously emerged among the UT26-derived hypermutator strain cells, and was highly expressed in these HYGO mutants, while no HYGO mutant appeared among UT26DAX-derived hypermutator strain cells, indicating the necessity of for the HYGO phenotype. His-tagged AdhX that was expressed in and purified to homogeneity showed ADH activity towards methanol and other alcohols. Mutagenesis analysis of the gene indicated a correlation between the ADH activity and the HYGO phenotype. These results demonstrated that the constitutive expression of an -encoding protein with ADH activity in UT26 leads to the CO-dependent HYGO phenotype. Identical or nearly identical orthologues were found in other sphingomonad strains, and most of them were located on plasmids, suggesting that the -mediated HYGO phenotype may be an important adaptation strategy to oligotrophic environments among sphingomonads.

Funding
This study was supported by the:
  • Yuji Nagata , Institute for Fermentation, Osaka (IFO)
  • Masataka Tsuda , Grants-in-Aid for Scientific Research (B) from JSPS , (Award 17H03781)
  • Yuji Nagata , Grants-in-Aid for Scientific Research (B) from JSPS , (Award 19H02865)
  • Yuji Nagata , Society of the Friendly Sons of St. Patrick for the Relief of Emigrants from Ireland , (Award no. 24658068, 26660054, and 16K14877)
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2020-04-20
2020-08-05
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