Biochemical and structural properties of a low-temperature-active glycoside hydrolase family 43 β-xylosidase: Activity and instability at high neutral salt concentrations

•A low-temperature-active glycoside hydrolase family 43 β-xylosidase was revealed.•The enzyme showed considerable activity and instability in high neutral salts.•Crystal structure comparison showed the enzyme had fewer salt bridges and H-bonds.•Structure of the enzyme was highly flexible with large positively charged surface.•Structural features led to low-temperature and high-salt adaptations of the enzyme. β-Xylosidase, of the glycoside hydrolase family 43 from Bacillus sp. HJ14, was expressed in Escherichia coli. Recombinant β-xylosidase (rHJ14GH43) exhibited maximum activity at 25 °C, approximately 15, 45, and 88% of maximum activity at 0, 10, and 20 °C, respectively, and poor stability at temperatures over 20 °C. rHJ14GH43 showed moderate or high activity, but poor stability, in NaCl, KCl, NaNO3, KNO3, Na2SO4, and (NH4)2SO4 at concentrations from 3.0 to 30.0% (w/v). The crystal structure of rHJ14GH43 was resolved and showed higher structural flexibility due to fewer salt bridges and hydrogen bonds compared to mesophilic and thermophilic β-xylosidases. High structural flexibility is presumed to be a key factor for catalytic adaptations to low temperatures and high salt concentrations. Approximately one-third of the surface of rHJ14GH43 is positively charged, which may be the primary factor responsible for poor stability in high neutral salt environments.