降解纤维素复合菌系F-7产纤维素酶的分离纯化及性质研究  

Purification and properties of a cellulase produced by fungus community F-7

作　　者：尚婷婷[1] 李全宏[2] 邓尚贵[1] 

机构地区：[1]浙江海洋学院食品与医药学院,舟山316000 [2]中国农业大学食品科学与营养工程学院,北京100083

出　　处：《食品科技》2015年第2期2-5,共4页Food Science and Technology

基　　金：国家重大科技合作项目(2012DFA30600);浙江省科技厅公益技术应用研究项目(2012C22063)

摘　　要：目的：对真菌复合菌系F-7所产纤维素酶进行分离纯化,并对其酶学性质进行研究。方法：通过（NH4）2SO4分级沉淀、透析、葡聚糖凝胶层析柱Sephadex G-75分离纯化技术,分离纯化来源于葡萄座腔菌属、米根霉、尖孢镰刀菌混合纤维素酶液中的内切葡萄糖苷酶。结果表明,CMC酶活在p H3.0~7.0的条件下,最适反应p H值为4.8,最适反应温度为50℃,超过60℃酶活迅速下降。结论：经凝胶层析柱Sephadex G-75和SDS-PAGE后第一个峰已纯化,得到了一种相对分子量约为66.2 ku的纤维素酶。分离纯化后该酶的比活力提高了1.75倍,回收率为20%。为进一步研究此纤维素酶的组分、结构和理化特性奠定了基础。Cellulase produced by F-7 cellulose-degrading microflora was isolated and purified, and its enzymatic properties were studied. The CMC-case were separated and purified from cellulase solutions produced by F-7 cellulose-degrading microflora was Botryosphaeria, Rhizopus oryzae and mixed enzymes of Fusarium oxysporum through（NH4）2SO4 fractional precipitation, dialysis and column chromatography of Sephadex G-75. The result showed, within the range of p H 3.0~7.0, the optimal reaction p H was 4.8. Higher enzyme activity appeared at 40~60 ℃, the optimal reaction temperature was at 50 ℃, and enzyme activity rapidly decreased when temperature was higher than 60 ℃. The protein was purified in first peak of Sephadex G-75 after SDS-PAGE, and its molecular mass was estimated as 66.2 ku. The specific activity of the cellulase increased 1.75 fold, and coefficient of recovery was 20%. It is the necessary preparing for the deep researching on cellulose.

关 键 词：纤维素酶 分离纯化 层析 凝胶过滤 

分 类 号：TS201.3[轻工技术与工程—食品科学]