6SC4
Gamma-Carbonic Anhydrase from the Haloarchaeon Halobacterium sp.
Summary for 6SC4
| Entry DOI | 10.2210/pdb6sc4/pdb |
| Related | 1V3W |
| Descriptor | Uncharacterized protein, ZINC ION, SULFATE ION, ... (8 entities in total) |
| Functional Keywords | red sea brine pool discovery deep, halophile, salt adaptation, enzyme engineering, metal binding protein |
| Biological source | candidate division MSBL1 archaeon SCGC-AAA259I09 |
| Total number of polymer chains | 5 |
| Total formula weight | 109026.91 |
| Authors | Vogler, M.,Karan, R.,Renn, D.,Vancea, A.,Vielberg, V.-T.,Groetzinger, S.W.,DasSarma, P.,Das Sarma, S.,Eppinger, J.,Groll, M.,Rueping, M. (deposition date: 2019-07-23, release date: 2020-04-22, Last modification date: 2024-01-24) |
| Primary citation | Vogler, M.,Karan, R.,Renn, D.,Vancea, A.,Vielberg, M.T.,Grotzinger, S.W.,DasSarma, P.,DasSarma, S.,Eppinger, J.,Groll, M.,Rueping, M. Crystal Structure and Active Site Engineering of a Halophilic gamma-Carbonic Anhydrase. Front Microbiol, 11:742-742, 2020 Cited by PubMed Abstract: Environments 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 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. PubMed: 32411108DOI: 10.3389/fmicb.2020.00742 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.6 Å) |
Structure validation
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