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 S. Al Rowaihi; Anastassja Akal; Wiebke Wackerow; John A. 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 S. Al Rowaihi, Anastassja Akal, Wiebke Wackerow, John A. Archer, Magnus Rueping, Dirk Weuster-Botz, Michael Groll, Jörg Eppinger, Stefan T. Arold^*

^*Corresponding author for this work

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

24 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 - Jan 19 2018

ASJC Scopus subject areas

Biochemistry
Molecular Medicine

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10.1021/acschembio.7b00792

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Grötzinger, S. W., Karan, R., Strillinger, E., Bader, S., Frank, A., Al Rowaihi, I. S., Akal, A., Wackerow, W., Archer, J. A., Rueping, M., Weuster-Botz, D., Groll, M., Eppinger, J., & Arold, S. T. (2018). 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 {"}microbial dark matter.{"}",

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

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doi = "10.1021/acschembio.7b00792",

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Grötzinger, SW, Karan, R, Strillinger, E, Bader, S, Frank, A, Al Rowaihi, IS, Akal, A, Wackerow, W, Archer, JA, Rueping, M, Weuster-Botz, D, Groll, M, Eppinger, J & Arold, ST 2018, '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

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AU - Grötzinger, Stefan W.

AU - Karan, Ram

AU - Strillinger, Eva

AU - Bader, Stefan

AU - Frank, Annika

AU - Al Rowaihi, Israa S.

AU - Akal, Anastassja

AU - Wackerow, Wiebke

AU - Archer, John A.

AU - Rueping, Magnus

AU - Weuster-Botz, Dirk

AU - Groll, Michael

AU - Eppinger, Jörg

AU - Arold, Stefan T.

PY - 2018/1/19

Y1 - 2018/1/19

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."

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Identification and Experimental Characterization of an Extremophilic Brine Pool Alcohol Dehydrogenase from Single Amplified Genomes

Abstract

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