通过(NH4)2SO4分级沉淀、HiPrep 26/10 Desalting脱盐柱、Source15Q阴离子交换柱、Source 15 S阳离子交换柱、HiTrap 16/60 Sephacryl S200 HR凝胶过滤等技术，分离纯化3种来源里氏木霉、黑曲霉、里氏木霉与黑曲霉混合纤维素酶液中的β-葡萄糖苷酶。结果表明，经SDSPAGE电泳鉴定均为电泳纯，测得黑氏木霉、黑曲霉单独培养β-葡萄糖苷酶相对分子质量分别为68、129 ku，而混合菌培养分离得到两种β-葡萄糖苷酶分子质量大小分别为66.2、134 ku。与单独培养的β-葡萄糖苷酶相似。3种来源的β-葡萄糖苷酶经多步分离纯化后的纯化倍数分别为37.25、40.21、30.12，酶活回收率分别为20.1 2%、23.21 %、28.56 %。
Abstract
The βglucosidase were separated and purified from cellulase solutions produced by Trichoderma reesei, Aspergillus niger, and mixed enzymes of T. reesei and A. niger through (NH4)2SO4 fractional precipitation, HiPrep 26/10 gel filtration, Source 15 Q and source 15 S ionexchange columns, and HiTrep 16/60 Sephacryl S200 HR gel column. Both β1,4glucosidase showed apparent homogeneity in SDSPAGE. The estimated molecular masses of β1,4glucosidase were 68 and 129 ku from T.reesei and A.niger, respectively. Two kinds of βglucosidases were purified from mixed culture of T.reesei and A.niger, two single subunit molecular masses were about 134 ku and 66.2 ku which are similar to the single culture. Three sources of βGA through multistep purification the purification multiple were 37.25, 40.21, 30.12 respectively. The recovery of enzyme activity were 20.12 %,23.21 %,28.56 %.
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基金
收稿日期：2010-07-19修回日期：2010-12-31 基金项目：国家自然科学基金项目（30871992）;江苏省科技支撑计划（BE2010732）；江苏高校科技创新团队资助项目作者简介：宋娜娜(1985—)，硕士生。*宋向阳（通信作者），副教授，博士。Email：xiangyangsong@hotmail.com。
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