Biochemical characteristics of a novel ethanol-tolerant xylanase from Bacillus subtilis subsp. subtilis JJBS250 and its applicability in saccharification of rice straw

Abstract

The present study reports the partial purification and characterisation of a novel xylanase from Bacillus subtilis subsp. subtilis JJBS250 followed by its applicability in the saccharification of rice straw. Bacterial xylanase was partially purified by gel filtration chromatography using Sephadax G-50 matrix that displayed optimal activity at 60 °C and pH 7.0 with half-life values of 1420 and 718 min at 60 °C and 80 °C, respectively. Bacterial xylanase showed high tolerance to ethanol with Km and Vmax values of 1.62 mg/ml and 23.92 U/ml, respectively, for beechwood xylan. Maximum amount of reducing sugars was obtained from sodium carbonate pretreated rice straw (151.65 mg/g substrate) as compared to untreated biomass (90.49 mg/g substrate) using 20 U/g enzyme dose at 60 °C and pH 7.0 after 48 h. Furthermore, the saccharification of pretreated rice straw was improved by synergistic action of bacterial and fungal xylanases. Analysis of rice straw using scanning electron microscopy (SEM), x-ray diffraction (XRD) and Fourier transform infrared (FTIR) spectroscopy clearly showed the morphological changes and delignification in rice straw after pretreatment. Bacterial xylanase exhibiting novel properties including ethanol tolerance and high thermostability is highly suitable for the production of second generation biofuels in simultaneous saccharification and fermentation process using lignocellulosic substrates.