毛栓菌木聚糖酶的纯化

Purification of Xylanase from Trametes trogii

[目的]进一步研究毛栓菌木聚糖酶的酶学性质。[方法]对毛栓菌木聚糖酶的粗酶液进行硫酸铵分级沉淀、DEAE-纤维素离子交换柱层析S、ephadexG100柱层析3步纯化,探讨木聚糖酶的产酶曲线及纯化方法。[结果]毛栓菌在最适培养基中培养时,木聚糖酶活力从第2天迅速提升,第8天达高峰,随后下降。经饱和度为30%～70%的硫酸铵分级沉淀后,酶活力回收率为84.931%,蛋白质含量为粗酶液的24.640%,纯化倍数为3.445。经DEAE-纤维素离子交换层析后,酶活力回收率为44.345%,酶蛋白纯化倍数为11.772。经SephadexG100柱层析后,酶活力回收率为20.201%,酶蛋白纯化倍数为16.123。[结论]毛栓菌木聚糖酶的粗酶液经过3步纯化后,酶活力回收率达20.201%,酶蛋白纯化倍数达16.123。

[Objective] The research aimed to further study the enzymology property of xylanase from Trametes trogii.[Method] The crude enzyme liquid of xylanase from T.trogii was made by the fractional precipitation of ammonium sulphate,EAE-cellulose ion-exchange chromatography and 3-step purification for Sephadex G100 column chromatography and the enzyme producing curve of xylanase and purification method were investigated.[Result] When T.trogii was cultured on the optimum medium,the xylanase activity was raised rapidly from the 2nd day,and reached to the peak on the 8th day,and then declined.After the fractional precipitation of ammonium sulphate with saturation degree of 30%-70%,the recovery rate of enzyme activity was 84.931%,the protein content was 24.640% of crude enzyme liquid,and purification fold was 3.445.After DEAE-cellulose ion-exchange chromatography,the enzyme activity recovery was 44.345%,and the apoenzyme purification fold was 11.772.After Sephadex G100 column chromatography,the enzyme activity recove ry was 20.201%,and the apoenzyme purification fold was 16.123.[Conclusion] After three-step purification on the crude enzyme of xylanase from T.trogii,the enzyme activity recovery was up to 20.201% and the apoenzyme purification fold was 16.123.