Purification and Characterization of a Thermostable Xylanase from Paenibacillus sp. NF1 and its Application in Xylooligosaccharides Production
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High levels of extracellular xylanase activity (211.79 IU/mg) produced by Paenibacillus sp. NF1were detected when it was submerged-cultured. After three consecutive purification stepsusing Octyl-Sepharose, Sephadex G75, and Q-Sepharose columns, a thermostable xylanase(XynNF) was purified to homogeneity and showed a molecular mass of 37 kDa according toSDS-PAGE. The specific activity of the purified XynNF was up to 3,081.05 IU/mg with a14.55-fold purification. The activity of XynNF was stimulated by Ca2+, Ba2+, DTT, andβ-mercaptoethanol, but was inhibited by Fe3+, Zn2+, Fe2+, Cu2+, SDS, and EDTA. The purifiedXynNF displayed a greater affinity for oat spelt xylan with the maximal enzymatic activity at60°C and pH 6.0. XynNF, which was shown to be cellulose-free, with high stability at hightemperature (70°C-80°C) and low pH range (pH 4.0-7.0), is potentially valuable for variousindustrial applications. The enzyme hydrolyzed oat spelt xylan to yield mainly xylooligosaccharides(95.8%) of 2-4 degree of polymerization (DP2-4). Moreover, the majority of the xylooligosacharides (DP2-4) products was xylobiose (61.5%). The thermostable xylanase (XynNF) thus seems potentiallyusefull in the production of xylooligosaccharides.
Abstract
High levels of extracellular xylanase activity (211.79 IU/mg) produced by Paenibacillus sp. NF1were detected when it was submerged-cultured. After three consecutive purification stepsusing Octyl-Sepharose, Sephadex G75, and Q-Sepharose columns, a thermostable xylanase(XynNF) was purified to homogeneity and showed a molecular mass of 37 kDa according toSDS-PAGE. The specific activity of the purified XynNF was up to 3,081.05 IU/mg with a14.55-fold purification. The activity of XynNF was stimulated by Ca2+, Ba2+, DTT, andβ-mercaptoethanol, but was inhibited by Fe3+, Zn2+, Fe2+, Cu2+, SDS, and EDTA. The purifiedXynNF displayed a greater affinity for oat spelt xylan with the maximal enzymatic activity at60°C and pH 6.0. XynNF, which was shown to be cellulose-free, with high stability at hightemperature (70°C-80°C) and low pH range (pH 4.0-7.0), is potentially valuable for variousindustrial applications. The enzyme hydrolyzed oat spelt xylan to yield mainly xylooligosaccharides(95.8%) of 2-4 degree of polymerization (DP2-4). Moreover, the majority of the xylooligosacharides (DP2-4) products was xylobiose (61.5%). The thermostable xylanase (XynNF) thus seems potentiallyusefull in the production of xylooligosaccharides.
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