Abstract
Xylanases have a broad range of applications in agro-industrial processes. In this study, we report on the discovery and characterization of a new thermotolerant GH10 xylanase from Bacillus safensis, designated as BsXyn10. The xylanase gene (bsxyn10) was cloned from Bacillus safensis and expressed in Escherichia coli. The reduced molecular mass of BsXyn10 was 48 kDa upon SDS-PAGE. Bsxyn10 was optimally active at pH 7.0 and 60 °C, stable over a broad range of pH (5.0-8.0), and also revealed tolerance toward different modulators (metal cations, EDTA). The enzyme was active toward various xylans with no activity on the glucose-based polysaccharides. KM, vmax, and kcat for oat spelt xylan hydrolysis were found to be 1.96 g·L-1, 58.6 μmole·min-1·(mg protein)-1, and 49 s-1, respectively. Thermodynamic parameters for oat spelt xylan hydrolysis at 60 °C were ΔS* = -61.9 J·mol-1·K-1, ΔH* = 37.0 kJ·mol-1 and ΔG* = 57.6 kJ·mol-1. BsXyn10 retained high levels of activity at temperatures up to 60 °C. The thermodynamic parameters (ΔH*D, ΔG*D, ΔS*D) for the thermal deactivation of BsXyn10 at a temperature range of 40-80 °C were: 192.5 ≤ ΔH*D ≤ 192.8 kJ·mol-1, 262.1 ≤ ΔS*D ≤ 265.8 J·mol-1·K-1, and 99.9 ≤ ΔG*D ≤ 109.6 kJ·mol-1. The BsXyn10-treated oat spelt xylan manifested the catalytic release of xylooligosaccharides of 2-6 DP, suggesting that BsXyn10 represents a promising candidate biocatalyst appropriate for several biotechnological applications.