Identification and experimental characterization of an extremophilic brine pool alcohol dehydrogenase from single amplified genomes

Handle URI:
http://hdl.handle.net/10754/626319
Title:
Identification and experimental characterization of an extremophilic brine pool alcohol dehydrogenase from single amplified genomes
Authors:
Grötzinger, Stefan W.; Karan, Ram; Strillinger, Eva; Bader, Stefan; Frank, Annika; Al Rowaihi, Israa ( 0000-0002-6953-8787 ) ; Akal, Anastassja; Wackerow, Wiebke; Archer, John A.C. ( 0000-0002-3302-3933 ) ; Rueping, Magnus ( 0000-0003-4580-5227 ) ; Weuster-Botz, Dirk; Groll, Michael; Eppinger, Jorg ( 0000-0001-7886-7059 ) ; Arold, Stefan T. ( 0000-0001-5278-0668 )
Abstract:
Because only 0.01% of prokaryotic genospecies can be cultured and in situ observations are often impracticable, culture-independent methods are required to understand microbial life and harness potential applications of microbes. Here, we report a methodology for the production of proteins with desired functions based on single amplified genomes (SAGs) from unculturable species. We use this method to resurrect an alcohol dehydrogenase (ADH/D1) from an uncharacterized halo-thermophilic archaeon collected from a brine pool at the bottom of the Red Sea. Our crystal structure of 5,6-dihydroxy NADPH-bound ADH/D1 combined with biochemical analyses reveal the molecular features of its halo-thermophily, its unique habitat adaptations, and its possible reaction mechanism for atypical oxygen activation. Our strategy offers a general guide for using SAGs as a source for scientific and industrial investigations of ‘microbial dark matter’.
KAUST Department:
Biological and Environmental Sciences and Engineering (BESE) Division; Computational Bioscience Research Center (CBRC); KAUST Catalysis Center (KCC); Physical Sciences and Engineering (PSE) Division
Citation:
Grötzinger SW, Karan R, Strillinger E, Bader S, Frank A, et al. (2017) Identification and experimental characterization of an extremophilic brine pool alcohol dehydrogenase from single amplified genomes. ACS Chemical Biology. Available: http://dx.doi.org/10.1021/acschembio.7b00792.
Publisher:
American Chemical Society (ACS)
Journal:
ACS Chemical Biology
KAUST Grant Number:
URF/1/1976.06; URF/1/2602.01.01; URF/1/1974
Issue Date:
30-Nov-2017
DOI:
10.1021/acschembio.7b00792
Type:
Article
ISSN:
1554-8929; 1554-8937
Sponsors:
The research reported in this publication was supported by the King Abdullah University of Science and Technology (KAUST) through the baseline fund and the Office of Sponsored Research (OSR) under award nos. URF/1/1976.06, URF/1/2602.01.01, and URF/1/1974. The authors are thankful for support for E.S. and S.G. from the International Graduate School of Science and Engineering (IGGSE), Technical University of Munich (TUM), project 8.03.
Additional Links:
http://pubs.acs.org/doi/abs/10.1021/acschembio.7b00792
Appears in Collections:
Articles; Physical Sciences and Engineering (PSE) Division; KAUST Catalysis Center (KCC); Computational Bioscience Research Center (CBRC); Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorGrötzinger, Stefan W.en
dc.contributor.authorKaran, Ramen
dc.contributor.authorStrillinger, Evaen
dc.contributor.authorBader, Stefanen
dc.contributor.authorFrank, Annikaen
dc.contributor.authorAl Rowaihi, Israaen
dc.contributor.authorAkal, Anastassjaen
dc.contributor.authorWackerow, Wiebkeen
dc.contributor.authorArcher, John A.C.en
dc.contributor.authorRueping, Magnusen
dc.contributor.authorWeuster-Botz, Dirken
dc.contributor.authorGroll, Michaelen
dc.contributor.authorEppinger, Jorgen
dc.contributor.authorArold, Stefan T.en
dc.date.accessioned2017-12-07T13:16:38Z-
dc.date.available2017-12-07T13:16:38Z-
dc.date.issued2017-11-30en
dc.identifier.citationGrötzinger SW, Karan R, Strillinger E, Bader S, Frank A, et al. (2017) Identification and experimental characterization of an extremophilic brine pool alcohol dehydrogenase from single amplified genomes. ACS Chemical Biology. Available: http://dx.doi.org/10.1021/acschembio.7b00792.en
dc.identifier.issn1554-8929en
dc.identifier.issn1554-8937en
dc.identifier.doi10.1021/acschembio.7b00792en
dc.identifier.urihttp://hdl.handle.net/10754/626319-
dc.description.abstractBecause only 0.01% of prokaryotic genospecies can be cultured and in situ observations are often impracticable, culture-independent methods are required to understand microbial life and harness potential applications of microbes. Here, we report a methodology for the production of proteins with desired functions based on single amplified genomes (SAGs) from unculturable species. We use this method to resurrect an alcohol dehydrogenase (ADH/D1) from an uncharacterized halo-thermophilic archaeon collected from a brine pool at the bottom of the Red Sea. Our crystal structure of 5,6-dihydroxy NADPH-bound ADH/D1 combined with biochemical analyses reveal the molecular features of its halo-thermophily, its unique habitat adaptations, and its possible reaction mechanism for atypical oxygen activation. Our strategy offers a general guide for using SAGs as a source for scientific and industrial investigations of ‘microbial dark matter’.en
dc.description.sponsorshipThe research reported in this publication was supported by the King Abdullah University of Science and Technology (KAUST) through the baseline fund and the Office of Sponsored Research (OSR) under award nos. URF/1/1976.06, URF/1/2602.01.01, and URF/1/1974. The authors are thankful for support for E.S. and S.G. from the International Graduate School of Science and Engineering (IGGSE), Technical University of Munich (TUM), project 8.03.en
dc.publisherAmerican Chemical Society (ACS)en
dc.relation.urlhttp://pubs.acs.org/doi/abs/10.1021/acschembio.7b00792en
dc.rightsThis document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Chemical Biology, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see http://pubs.acs.org/doi/abs/10.1021/acschembio.7b00792.en
dc.subjectHalophilicen
dc.subjectthermophilicen
dc.subjectextremozymeen
dc.subjectsingle.cell amplified genomeen
dc.subject5,6.dihydroxy NADPHen
dc.subjectbiotechnologyen
dc.titleIdentification and experimental characterization of an extremophilic brine pool alcohol dehydrogenase from single amplified genomesen
dc.typeArticleen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.contributor.departmentComputational Bioscience Research Center (CBRC)en
dc.contributor.departmentKAUST Catalysis Center (KCC)en
dc.contributor.departmentPhysical Sciences and Engineering (PSE) Divisionen
dc.identifier.journalACS Chemical Biologyen
dc.eprint.versionPost-printen
dc.contributor.institutionTechnical University of Munich, Department of Mechanical Engineering, Institute of Biochemical Engineering, Garching, Germanyen
dc.contributor.institutionTechnical University of Munich, Center for Integrated Protein Science Munich in the Department Chemistry, Garching, Germanyen
kaust.authorGrötzinger, Stefan W.en
kaust.authorKaran, Ramen
kaust.authorStrillinger, Evaen
kaust.authorBader, Stefanen
kaust.authorAl Rowaihi, Israaen
kaust.authorAkal, Anastassjaen
kaust.authorWackerow, Wiebkeen
kaust.authorArcher, John A.C.en
kaust.authorRueping, Magnusen
kaust.authorEppinger, Jorgen
kaust.authorArold, Stefan T.en
kaust.grant.numberURF/1/1976.06en
kaust.grant.numberURF/1/2602.01.01en
kaust.grant.numberURF/1/1974en
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