Expression of an alcohol dehydrogenase gene in a heterotrophic bacterium induces carbon dioxide-dependent high-yield growth under oligotrophic conditions
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 isola...
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Veröffentlicht in: | Microbiology (Society for General Microbiology) 2020-06, Vol.166 (6), p.531-545 |
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creator | Inaba, Shinnosuke Sakai, Hironori Kato, Hiromi Horiuchi, Takayuki Yano, Hirokazu Ohtsubo, Yoshiyuki Tsuda, Masataka Nagata, Yuji |
description | 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. |
doi_str_mv | 10.1099/mic.0.000908 |
format | Article |
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-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.</description><identifier>ISSN: 1350-0872</identifier><identifier>EISSN: 1465-2080</identifier><identifier>DOI: 10.1099/mic.0.000908</identifier><identifier>PMID: 32310743</identifier><language>eng</language><publisher>England</publisher><subject>Alcohol Dehydrogenase - genetics ; Alcohol Dehydrogenase - metabolism ; Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Carbon Dioxide - metabolism ; Gene Expression Regulation, Bacterial ; Heterotrophic Processes ; Hexachlorocyclohexane - metabolism ; Mutation ; Phenotype ; Plasmids - genetics ; Plasmids - metabolism ; Promoter Regions, Genetic ; Sphingomonadaceae - enzymology ; Sphingomonadaceae - genetics ; Sphingomonadaceae - growth & development ; Sphingomonadaceae - metabolism</subject><ispartof>Microbiology (Society for General Microbiology), 2020-06, Vol.166 (6), p.531-545</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c395t-fb26c32c80d3719f30917fa0798c90a4517cf6d96f87dd38ae9b4abca9f7fc873</citedby><cites>FETCH-LOGICAL-c395t-fb26c32c80d3719f30917fa0798c90a4517cf6d96f87dd38ae9b4abca9f7fc873</cites><orcidid>0000-0002-5388-663X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,781,785,27926,27927</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32310743$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Inaba, Shinnosuke</creatorcontrib><creatorcontrib>Sakai, Hironori</creatorcontrib><creatorcontrib>Kato, Hiromi</creatorcontrib><creatorcontrib>Horiuchi, Takayuki</creatorcontrib><creatorcontrib>Yano, Hirokazu</creatorcontrib><creatorcontrib>Ohtsubo, Yoshiyuki</creatorcontrib><creatorcontrib>Tsuda, Masataka</creatorcontrib><creatorcontrib>Nagata, Yuji</creatorcontrib><title>Expression of an alcohol dehydrogenase gene in a heterotrophic bacterium induces carbon dioxide-dependent high-yield growth under oligotrophic conditions</title><title>Microbiology (Society for General Microbiology)</title><addtitle>Microbiology (Reading)</addtitle><description>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.</description><subject>Alcohol Dehydrogenase - genetics</subject><subject>Alcohol Dehydrogenase - metabolism</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>Carbon Dioxide - metabolism</subject><subject>Gene Expression Regulation, Bacterial</subject><subject>Heterotrophic Processes</subject><subject>Hexachlorocyclohexane - metabolism</subject><subject>Mutation</subject><subject>Phenotype</subject><subject>Plasmids - genetics</subject><subject>Plasmids - metabolism</subject><subject>Promoter Regions, Genetic</subject><subject>Sphingomonadaceae - enzymology</subject><subject>Sphingomonadaceae - genetics</subject><subject>Sphingomonadaceae - growth & development</subject><subject>Sphingomonadaceae - metabolism</subject><issn>1350-0872</issn><issn>1465-2080</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kMtOwzAQRS0EoqWwY438AaSM4yaOl6gqD6kSG1hHjj1ujNI4slPRfgp_i1Ghq5nRuboaHUJuGcwZSPmwdXoOcwCQUJ2RKVuURZZDBedp5wVkUIl8Qq5i_ARIENglmfCcMxALPiXfq_0QMEbne-otVT1Vnfat76jB9mCC32CvItI0kLpEaYsjBj8GP7RO00bpdLrdNkGz0xipVqFJZcb5vTOYGRywN9iPtHWbNjs47AzdBP81tnSXQKC-c5tTn_a9cWP6Jl6TC6u6iDd_c0Y-nlbvy5ds_fb8unxcZ5rLYsxsk5ea57oCwwWTloNkwioQstIS1KJgQtvSyNJWwhheKZTNQjVaSSusrgSfkftjrw4-xoC2HoLbqnCoGdS_hutkuIb6aDjF747xYdds0ZzC_0r5Dwc5fBo</recordid><startdate>20200601</startdate><enddate>20200601</enddate><creator>Inaba, Shinnosuke</creator><creator>Sakai, Hironori</creator><creator>Kato, Hiromi</creator><creator>Horiuchi, Takayuki</creator><creator>Yano, Hirokazu</creator><creator>Ohtsubo, Yoshiyuki</creator><creator>Tsuda, Masataka</creator><creator>Nagata, Yuji</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><orcidid>https://orcid.org/0000-0002-5388-663X</orcidid></search><sort><creationdate>20200601</creationdate><title>Expression of an alcohol dehydrogenase gene in a heterotrophic bacterium induces carbon dioxide-dependent high-yield growth under oligotrophic conditions</title><author>Inaba, Shinnosuke ; Sakai, Hironori ; Kato, Hiromi ; Horiuchi, Takayuki ; Yano, Hirokazu ; Ohtsubo, Yoshiyuki ; Tsuda, Masataka ; Nagata, Yuji</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c395t-fb26c32c80d3719f30917fa0798c90a4517cf6d96f87dd38ae9b4abca9f7fc873</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Alcohol Dehydrogenase - genetics</topic><topic>Alcohol Dehydrogenase - metabolism</topic><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - metabolism</topic><topic>Carbon Dioxide - metabolism</topic><topic>Gene Expression Regulation, Bacterial</topic><topic>Heterotrophic Processes</topic><topic>Hexachlorocyclohexane - metabolism</topic><topic>Mutation</topic><topic>Phenotype</topic><topic>Plasmids - genetics</topic><topic>Plasmids - metabolism</topic><topic>Promoter Regions, Genetic</topic><topic>Sphingomonadaceae - enzymology</topic><topic>Sphingomonadaceae - genetics</topic><topic>Sphingomonadaceae - growth & development</topic><topic>Sphingomonadaceae - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Inaba, Shinnosuke</creatorcontrib><creatorcontrib>Sakai, Hironori</creatorcontrib><creatorcontrib>Kato, Hiromi</creatorcontrib><creatorcontrib>Horiuchi, Takayuki</creatorcontrib><creatorcontrib>Yano, Hirokazu</creatorcontrib><creatorcontrib>Ohtsubo, Yoshiyuki</creatorcontrib><creatorcontrib>Tsuda, Masataka</creatorcontrib><creatorcontrib>Nagata, Yuji</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><jtitle>Microbiology (Society for General Microbiology)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Inaba, Shinnosuke</au><au>Sakai, Hironori</au><au>Kato, Hiromi</au><au>Horiuchi, Takayuki</au><au>Yano, Hirokazu</au><au>Ohtsubo, Yoshiyuki</au><au>Tsuda, Masataka</au><au>Nagata, Yuji</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression of an alcohol dehydrogenase gene in a heterotrophic bacterium induces carbon dioxide-dependent high-yield growth under oligotrophic conditions</atitle><jtitle>Microbiology (Society for General Microbiology)</jtitle><addtitle>Microbiology (Reading)</addtitle><date>2020-06-01</date><risdate>2020</risdate><volume>166</volume><issue>6</issue><spage>531</spage><epage>545</epage><pages>531-545</pages><issn>1350-0872</issn><eissn>1465-2080</eissn><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.</abstract><cop>England</cop><pmid>32310743</pmid><doi>10.1099/mic.0.000908</doi><tpages>15</tpages><orcidid>https://orcid.org/0000-0002-5388-663X</orcidid><oa>free_for_read</oa></addata></record> |
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source | MEDLINE; PubMed Central |
subjects | Alcohol Dehydrogenase - genetics Alcohol Dehydrogenase - metabolism Bacterial Proteins - genetics Bacterial Proteins - metabolism Carbon Dioxide - metabolism Gene Expression Regulation, Bacterial Heterotrophic Processes Hexachlorocyclohexane - metabolism Mutation Phenotype Plasmids - genetics Plasmids - metabolism Promoter Regions, Genetic Sphingomonadaceae - enzymology Sphingomonadaceae - genetics Sphingomonadaceae - growth & development Sphingomonadaceae - metabolism |
title | Expression of an alcohol dehydrogenase gene in a heterotrophic bacterium induces carbon dioxide-dependent high-yield growth under oligotrophic conditions |
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