Show simple item record

dc.contributor.authorVogler, Malvina M.
dc.contributor.authorKaran, Ram
dc.contributor.authorRenn, Dominik
dc.contributor.authorVancea, Alexandra
dc.contributor.authorVielberg, Marie-Theres
dc.contributor.authorGrötzinger, Stefan W.
dc.contributor.authorDasSarma, Priya
dc.contributor.authorDasSarma, Shiladitya
dc.contributor.authorEppinger, Jörg
dc.contributor.authorGroll, Michael
dc.contributor.authorRueping, Magnus
dc.identifier.citationVogler, M., Karan, R., Renn, D., Vancea, A., Vielberg, M.-T., Grötzinger, S. W., … Rueping, M. (2020). Crystal Structure and Active Site Engineering of a Halophilic γ-Carbonic Anhydrase. Frontiers in Microbiology, 11. doi:10.3389/fmicb.2020.00742
dc.description.abstractEnvironments previously thought to be uninhabitable offer a tremendous wealth of unexplored microorganisms and enzymes. In this paper, we present the discovery and characterization of a novel γ-carbonic anhydrase (γ-CA) from the polyextreme Red Sea brine pool Discovery Deep (2141 m depth, 44.8°C, 26.2% salt) by single-cell genome sequencing. The extensive analysis of the selected gene helps demonstrate the potential of this culture-independent method. The enzyme was expressed in the bioengineered haloarchaeon Halobacterium sp. NRC-1 and characterized by X-ray crystallography and mutagenesis. The 2.6 Å crystal structure of the protein shows a trimeric arrangement. Within the γ-CA, several possible structural determinants responsible for the enzyme’s salt stability could be highlighted. Moreover, the amino acid composition on the protein surface and the intra- and intermolecular interactions within the protein differ significantly from those of its close homologs. To gain further insights into the catalytic residues of the γ-CA enzyme, we created a library of variants around the active site residues and successfully improved the enzyme activity by 17-fold. As several γ-CAs have been reported without measurable activity, this provides further clues as to critical residues. Our study reveals insights into the halophilic γ-CA activity and its unique adaptations. The study of the polyextremophilic carbonic anhydrase provides a basis for outlining insights into strategies for salt adaptation, yielding enzymes with industrially valuable properties, and the underlying mechanisms of protein evolution.
dc.description.sponsorshipThe research reported in this publication was supported by the King Abdullah University of Science and Technology (KAUST), Kingdom of Saudi Arabia. The staff of the Beamline X06SA at the Paul Scherrer Institute, SLS, Villigen, Switzerland, is acknowledged for assistance during data collection. Work in the DasSarma laboratory was supported by grant 80NSSC19K0463.
dc.publisherFrontiers Media SA
dc.rightsThis is an open access article.
dc.titleCrystal Structure and Active Site Engineering of a Halophilic γ-Carbonic Anhydrase.
dc.contributor.departmentAcademic Affairs
dc.contributor.departmentBiological & Organometallic Catalysis Laboratories
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Division
dc.contributor.departmentBioscience Program
dc.contributor.departmentChemical Science Program
dc.contributor.departmentKAUST Catalysis Center (KCC)
dc.contributor.departmentOffice of the VP
dc.contributor.departmentPhysical Science and Engineering (PSE) Division
dc.identifier.journalFrontiers in microbiology
dc.eprint.versionPublisher's Version/PDF
kaust.personVogler, Malvina
kaust.personKaran, Ram
kaust.personVancea, Alexandra-Ioana
kaust.personVielberg, Marie-Theres
kaust.personGrötzinger, Stefan W.
kaust.personDasSarma, Priya
kaust.personDasSarma, Shiladitya
kaust.personEppinger, Jorg
kaust.personGroll, Michael
kaust.personRueping, Magnus

Files in this item

Publisher's Version/PDF

This item appears in the following Collection(s)

Show simple item record

This is an open access article.
Except where otherwise noted, this item's license is described as This is an open access article.