Molecular cloning, expression and characterization of β-xylanase from Thermotoga Naphthophila for bioethanol production of bioethanol

Enzymes can be used in various biotechnological applications due to the easy and cheap production stage. Since xylanase enzymes are preferrend especially in industrial areas, researches on this enzyme are quite high. In this study, the β-xylanase gene was cloned from Thermotoga naphthophila, a thermophilic organism. The expression vector was expressed in Escherichia coli BL21(DE3) via pE-T21a (+). Within the literature information, pH (7.0), temperature (37 °C) and IPTG concentration (0,5 mM) of the enzyme were adjusted to result in the highest expression. Dinitrosalicylic acid (DNS) was used to determine the amount of reducing sugar of the enzyme. The molecular mass of the purified β-xylanase enzyme was found to be 38 kDa using sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. The β-xylanase enzyme hazelnut cob, rhododendron branch and rhododendron leaf were pretreated with lignocellulosic biomass. Many pretreatment methods have been researched and the acid alkali method which can give the best results, has been used. The amount of sugar released by enzyme treatment of biomass was calculated. The highest activity was observed in hazelnut cob biomass. It is envisaged that after the thermophilic β-xylanase enzyme, which has various uses, is treated with biomass, fermentation stage and other methods can replace fossil fuels and produce bioethanol as a renewable energy source.