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

Stefan W. Grötzinger; Ram Karan; Eva Strillinger; Stefan Bader; Annika Frank; Israa Al Rowaihi; Anastassja Gespers (Akal); Wiebke Wackerow; John A.C. Archer; Magnus Rueping; Dirk Weuster-Botz; Michael Groll; Jörg Eppinger; Stefan T. Arold

doi:10.1021/acschembio.7b00792

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

Stefan W. Grötzinger, Ram Karan, Eva Strillinger, Stefan Bader, Annika Frank, Israa Al Rowaihi, Anastassja Gespers (Akal), Wiebke Wackerow, John A.C. Archer, Magnus Rueping, Dirk Weuster-Botz, Michael Groll, Jörg Eppinger, Stefan T. Arold

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

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’.

Original language	English (US)
Pages (from-to)	161-170
Number of pages	10
Journal	ACS Chemical Biology
Volume	13
Issue number	1
DOIs	https://doi.org/10.1021/acschembio.7b00792
State	Published - Dec 18 2017

Bibliographical note

KAUST Repository Item: Exported on 2020-10-01
Acknowledged KAUST grant number(s): URF/1/1976.06, URF/1/2602.01.01, URF/1/1974
Acknowledgements: 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.

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https://repository.kaust.edu.sa/bitstream/10754/626319/1/acschembio.7b00792.pdf

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Grötzinger, S. W., Karan, R., Strillinger, E., Bader, S., Frank, A., Al Rowaihi, I., Gespers (Akal), A., Wackerow, W., Archer, J. A. C., Rueping, M., Weuster-Botz, D., Groll, M., Eppinger, J., & Arold, S. T. (2017). Identification and experimental characterization of an extremophilic brine pool alcohol dehydrogenase from single amplified genomes. ACS Chemical Biology, 13(1), 161-170. https://doi.org/10.1021/acschembio.7b00792

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title = "Identification and experimental characterization of an extremophilic brine pool alcohol dehydrogenase from single amplified genomes",

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 {\textquoteleft}microbial dark matter{\textquoteright}.",

author = "Gr{\"o}tzinger, {Stefan W.} and Ram Karan and Eva Strillinger and Stefan Bader and Annika Frank and {Al Rowaihi}, Israa and {Gespers (Akal)}, Anastassja and Wiebke Wackerow and Archer, {John A.C.} and Magnus Rueping and Dirk Weuster-Botz and Michael Groll and J{\"o}rg Eppinger and Arold, {Stefan T.}",

note = "KAUST Repository Item: Exported on 2020-10-01 Acknowledged KAUST grant number(s): URF/1/1976.06, URF/1/2602.01.01, URF/1/1974 Acknowledgements: 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.",

year = "2017",

month = dec,

day = "18",

doi = "10.1021/acschembio.7b00792",

language = "English (US)",

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Grötzinger, SW, Karan, R, Strillinger, E, Bader, S, Frank, A, Al Rowaihi, I, Gespers (Akal), A, Wackerow, W, Archer, JAC, Rueping, M, Weuster-Botz, D, Groll, M, Eppinger, J & Arold, ST 2017, 'Identification and experimental characterization of an extremophilic brine pool alcohol dehydrogenase from single amplified genomes', ACS Chemical Biology, vol. 13, no. 1, pp. 161-170. https://doi.org/10.1021/acschembio.7b00792

TY - JOUR

T1 - Identification and experimental characterization of an extremophilic brine pool alcohol dehydrogenase from single amplified genomes

AU - Grötzinger, Stefan W.

AU - Karan, Ram

AU - Strillinger, Eva

AU - Bader, Stefan

AU - Frank, Annika

AU - Al Rowaihi, Israa

AU - Gespers (Akal), Anastassja

AU - Wackerow, Wiebke

AU - Archer, John A.C.

AU - Rueping, Magnus

AU - Weuster-Botz, Dirk

AU - Groll, Michael

AU - Eppinger, Jörg

AU - Arold, Stefan T.

N1 - KAUST Repository Item: Exported on 2020-10-01 Acknowledged KAUST grant number(s): URF/1/1976.06, URF/1/2602.01.01, URF/1/1974 Acknowledgements: 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.

PY - 2017/12/18

Y1 - 2017/12/18

N2 - 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’.

AB - 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’.

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AN - SCOPUS:85040867318

SN - 1554-8929

VL - 13

SP - 161

EP - 170

JO - ACS Chemical Biology

JF - ACS Chemical Biology

IS - 1

ER -

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

Abstract

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