High-activity production of xylanase by Pichia stipitis: Purification, characterization, kinetic evaluation and xylooligosaccharides production

As an industrially important biological macromolecule, xylanase hydrolyzes xylan to produce xylooligosaccarides (XOS). XOS, with a degree of polymerization (DP) 2 to 4, are important prebiotics used as food ingredients. In this study, xylanase (5536 U/g substrate) was produced by Pichia stipitis using corncob and wheat bran mixture under solid state fermentation. Crude xylanase were purified and biochemically characterized. XOS hydrolyzed by crude and purified xylanases were quantified. Molecular weight of the purified enzyme was around 31.6 kDa on SDS-PAGE. Enzyme kinetics showed Km and Vmax values of 4.52 mg/mL and 9.17 μmol/min/mL, respectively. The optimal conditions were pH 6.0 and 50 °C. Xylanase was stable at pH 5-8 for 60 min by retaining 57% activity and at 50 °C for 80 min by retaining 65% activity. Cooper and potassium had no inhibitory effect on xylanase activity. Xylan hydrolysates produced by purified xylanase contained 92% XOS consisting of 14% xylotetroase (DP 4), 49% xylotriose (DP 3) and 29% xylobiose (DP 2). These findings indicate the potential of applying purified xylanase for industrial XOS production.