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使用硫酸铵分级沉淀、SephadexG 25凝胶色谱脱盐、DEAE SephadexA 50和SP SephadexC 50离子交换色谱等分离纯化技术,从里氏木霉(Trichodermareesei)RutC 30培养液中分离出木聚糖酶组分,再经SephadexG 100凝胶过滤色谱进一步分离纯化,得到2个纯木聚糖酶组分A和组分B。经SDS PAGE鉴定两组分为单带,相对分子质量分别为20300和13500。组分A的最适反应条件为45℃、pH4.5,在pH3.0～5.5很稳定,酶解产物主要是低聚木糖,只含少量木糖;组分B的最适反应条件为55℃、pH5.5,酶解产物全部是低聚木糖。
The components of xylanase were isolated from the Trichoderma reesei RutC 30 medium using ammonium sulfate fractionation, SephadexG 25 gel desalting, DEAE Sephadex A 50 and SP Sephadex C 50 ion-exchange chromatography. After further purification by Sephadex G 100 gel filtration chromatography, two pure xylanase components A and B were obtained. SDS PAGE identified two components as a single band, the relative molecular mass of 20300 and 13500 respectively. The optimal reaction conditions of component A were 45 ℃, pH4.5, stable at pH3.0 ~ 5.5, and the hydrolyzate was mainly xylooligosaccharides with only a small amount of xylose. The optimum reaction conditions of component B were 55 ° C, pH5.5, all the hydrolyzate xylooligosaccharides.