[1]宋娜娜,宋向阳*,连之娜,等.纤维素酶液中β-葡萄糖苷酶的分离纯化[J].南京林业大学学报(自然科学版),2011,35(04):111-116.[doi:10.3969/j.jssn.1000-2006.2011.04.023]
 SONG Nana,SONG Xiangyang*,LIAN Zhina,et al.Isolation and purification of β-glucosidase from cellulase solution[J].Journal of Nanjing Forestry University(Natural Science Edition),2011,35(04):111-116.[doi:10.3969/j.jssn.1000-2006.2011.04.023]
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纤维素酶液中β-葡萄糖苷酶的分离纯化
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《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

卷:
35
期数:
2011年04期
页码:
111-116
栏目:
研究论文
出版日期:
2011-07-31

文章信息/Info

Title:
Isolation and purification of β-glucosidase from cellulase solution
作者:
宋娜娜宋向阳*连之娜勇强余世袁
南京林业大学化学工程学院,林木遗传与生物技术省部共建教育部重点实验室,江苏南京210037
Author(s):
SONG NanaSONG Xiangyang*LIAN ZhinaYONG QiangYU Shiyuan
Key Laboratory of Forest Genetics & Biotechnology,Ministry of Education,College of Chemical Engineering, Nanjing Forestry University, Nanjing,210037,China
关键词:
里氏木霉黑曲霉纤维素酶β-葡萄糖苷酶分离纯化
Keywords:
Trichoderma reesei Aspergillus nigercellulase βglucosidaseisolation and purification
分类号:
Q814
DOI:
10.3969/j.jssn.1000-2006.2011.04.023
文献标志码:
A
摘要:
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 S200 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SDSPAGE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 βglucosidase 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 ionexchange columns, and HiTrep 16/60 Sephacryl S200 HR gel column. Both β1,4glucosidase showed apparent homogeneity in SDSPAGE. The estimated molecular masses of β1,4glucosidase were 68 and 129 ku from T.reesei and A.niger, respectively. Two kinds of βglucosidases 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 βGA through multistep 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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备注/Memo

备注/Memo:
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。Email:xiangyangsong@hotmail.com。
更新日期/Last Update: 2011-08-13