开普曲霉来源的低分子量木聚糖酶(AcXyn)的纯化与性质研究

    Purification and characterization of a xylanase (AcXyn) from Aspergillus capensis

    • 摘要: 为挖掘对阿拉伯木聚糖具有高效水解活性的木聚糖酶,通过对土壤中天然微生物筛选,得到一株降解阿拉伯木聚糖的菌株,鉴定为开普曲霉(Aspergillus capensis),命名为FAHD19。该菌株发酵液经硫酸铵沉淀和强阴离子交换层析两步纯化,得到电泳级纯酶(AcXyn)并对其酶学性质进行表征。结果表明:木聚糖酶AcXyn分子量为16.6 kDa;最适pH为6.0,在pH值 5.0~11.5范围内有良好的稳定性;最适温度为55 ℃,在50 ℃以下具有良好的稳定性;对多种金属离子和化合物具有良好的耐受性,且具有严格的木聚糖底物特异性,对小麦阿拉伯木聚糖的比酶活力为367.9 U/mg,能将其水解为聚合度4~6的低聚木糖。开普曲霉所产木聚糖酶AcXyn具有优异的酶学性质和良好的降解阿拉伯木聚糖的能力,其酶学性质契合烘焙工艺,在食品加工领域具有潜在的应用价值。

       

      Abstract: Arabinoxylan is found in the cell wall of a wide variety of cereal. Arabinoxylan in wheat flour could compete with the gluten protein for water absorption in the dough, which would increase the hardness of the dough. The xylanase is a key enzyme in the xylanolytic enzyme system and is widely applied in industries including pulp and paper, animal feed, beverages and food. However, the complex structure of arabinoxylan makes most xylanases have limited activity to degrade it. In order to discover xylanase with high arabinoxylan hydrolysing activity, we screened arabinoxylan-degrading microorganisms from soil. We screened for a fungus, designated FAHD19, was identified as Aspergillus capensis. The xylanase from Aspergillus capensis, named AcXyn, was purified to electrophoretic homogeneity through ammonium sulfate precipitation and strong anion-exchange chromatography. We characterised its enzymatic properties and the results showed that the molecular weight of AcXyn was 16.6 kDa. The purified xylanase showed the optimal temperature of 55 ℃, and pH 6.0 with a stable xylanolytic activity within the temperature range of 25-50 ℃, and within the pH range of 5.0-11.5. AcXyn demonstrated good tolerance to various metal ions and chemicals. The presence of Co2+ enhanced xylanase activity. but SDS showed inhibition of xylanase activity. It exhibited strict substrate specificity, and had the highest specific activity of 367.9 U/mg towards wheat arabinoxylan. Hydrolysis property analysis revealed that AcXyn degraded arabinoxylan into low-molecular-weight xylooligosaccharides with degrees of polymerization from 4 to 6. AcXyn hydrolyzed xylotetraose to xylooligosaccharides(XOSs) with lower degree of polymerizations(DPs) than itself, but could not hydrolyze xylobiose and xylotriose. In this study, we reported a new xylanase-producing strain that enriches the source of xylanase from Aspergillus species. The AcXyn produced by Aspergillus capensis had excellent arabinoxylan degradation capacity and enzymatic properties suitable for baking processes, which made it potentially valuable for applications in food processing.

       

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