4qqs | pdb_00004qqs
Crystal structure of a thermostable family-43 glycoside hydrolase
Structural highlights
Publication Abstract from PubMedA gene from the heterotrophic, halothermophilic marine bacterium Halothermothrix orenii has been cloned and overexpressed in Escherichia coli. This gene encodes the only glycoside hydrolase of family 43 (GH43) produced by H. orenii. The crystal structure of the H. orenii glycosidase was determined by molecular replacement and refined at 1.10 A resolution. As for other GH43 members, the enzyme folds as a five-bladed beta-propeller. The structure features a metal-binding site on the propeller axis, near the active site. Based on thermal denaturation data, the H. orenii glycosidase depends on divalent cations in combination with high salt for optimal thermal stability against unfolding. A maximum melting temperature of 76 degrees C was observed in the presence of 4 M NaCl and Mn(2+) at pH 6.5. The gene encoding the H. orenii GH43 enzyme has previously been annotated as a putative alpha-L-arabinofuranosidase. Activity was detected with p-nitrophenyl-alpha-L-arabinofuranoside as a substrate, and therefore the name HoAraf43 was suggested for the enzyme. In agreement with the conditions for optimal thermal stability against unfolding, the highest arabinofuranosidase activity was obtained in the presence of 4 M NaCl and Mn(2+) at pH 6.5, giving a specific activity of 20-36 micromol min(-1) mg(-1). The active site is structurally distinct from those of other GH43 members, including arabinanases, arabinofuranosidases and xylanases. This probably reflects the special requirements for degrading the unique biomass available in highly saline aqueous ecosystems, such as halophilic algae and halophytes. The amino-acid distribution of HoAraf43 has similarities to those of mesophiles, thermophiles and halophiles, but also has unique features, for example more hydrophobic amino acids on the surface and fewer buried charged residues. High-resolution crystal structure of a polyextreme GH43 glycosidase from Halothermothrix orenii with alpha-L-arabinofuranosidase activity.,Hassan N, Kori LD, Gandini R, Patel BK, Divne C, Tan TC Acta Crystallogr F Struct Biol Commun. 2015 Mar 1;71(Pt 3):338-45. doi:, 10.1107/S2053230X15003337. Epub 2015 Feb 19. PMID:25760712[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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