【摘要】 黑曲霉(Aspergillus niger)WM20-11固态发酵成熟曲,经磷酸缓冲液浸提、硫酸铵分步盐析、DEAE-Sepharose fast flow阴离子交换层析、Sephadex G-100凝胶过滤层析等分离纯化手段,最终获得了Native-PAGE、SDS-PAGE纯的酸性β-甘露聚糖酶组分,其纯化倍数为25.08,收率为5.1%。Sephadex G-100凝胶过滤层析和SDS-PAGE测得纯酶的相对分子质量分别为39 kD和40 kD,表明该酶以单体形式存在;IEF-PAGE测得该酶的等电点为4.0;含糖量测定为19.6%;酶蛋白氨基酸组成中(Asp+Glu)/(Lys+Arg)为3.74。更多还原

【Abstract】 The acidic β-mannanase from Aspergillus niger WM20-11 was purified by buffer extraction,ammonium sulfate precipitation,DEAE-Sepharose fast flow and Sephadex G-100 column chromatographies. The purified enzyme was homogeneous on Native-PAGE and SDS-PAGE.After these steps the enzyme was purified by 25.08-fold with a recovery of 5.1%.Molecular weight of the β-mannanase was determined as 39 kD on Sephadex G-100 gel filtration and 40 kD on SDS-PAGE,which indicated that the β-mannanase was a monomer.The isoelectric point was estimated to be 4.0 by IEF-PAGE.Its carbohydrate content was 19.6%.The ratio of(Asp+Glu)/(Lys+Arg) was 3.74.更多还原