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’.
UR - http://hdl.handle.net/10754/626319
UR - http://pubs.acs.org/doi/abs/10.1021/acschembio.7b00792
UR - http://www.scopus.com/inward/record.url?scp=85040867318&partnerID=8YFLogxK
U2 - 10.1021/acschembio.7b00792
DO - 10.1021/acschembio.7b00792
M3 - Article
C2 - 29188989
AN - SCOPUS:85040867318
SN - 1554-8929
VL - 13
SP - 161
EP - 170
JO - ACS Chemical Biology
JF - ACS Chemical Biology
IS - 1
ER -