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[1]唐德芳,裴小琼,李晓璐,等.黑曲霉β-葡萄糖苷酶的筛选、克隆及表达[J].应用与环境生物学报,2009,15(03):423-426.[doi:10.3724/SP.J.1145.2009.00423]
 TANG Defang,PEI Xiaoqiong,LI Xiaolu,et al.Screening, Cloning and Expression of Aspergillus niger β-glucosidase[J].Chinese Journal of Applied & Environmental Biology,2009,15(03):423-426.[doi:10.3724/SP.J.1145.2009.00423]
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黑曲霉β-葡萄糖苷酶的筛选、克隆及表达()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
15卷
期数:
2009年03期
页码:
423-426
栏目:
生物质能源专辑
出版日期:
2009-05-15

文章信息/Info

Title:
Screening, Cloning and Expression of Aspergillus niger β-glucosidase
作者:
唐德芳裴小琼李晓璐胡承陈军吴中柳
1中国科学院成都生物研究所 成都 610041
2四川大学生命科学学院生物资源与生态环境教育部重点实验室 成都 610041
3成都医学院科研实验中心 成都 610083
Author(s):
TANG DefangPEI XiaoqiongLI XiaoluHU ChengCHEN JunWU Zhongliu
1Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China
2MOE Laboratory of Bioresource and Environment, College of Life Sciences, Sichuan University, Chengdu 610041, China
3Scientific Research Center, Chengdu Medical College, Chengdu 610083, China
关键词:
黑曲霉β-葡萄糖苷酶克隆RT-PCR毕赤酵母表达
Keywords:
Aspergillus niger β-glucosidase gene cloning RT-PCR Pichia pastoris expression
分类号:
Q936 + Q556.2
DOI:
10.3724/SP.J.1145.2009.00423
文献标志码:
A
摘要:
摘 要 从曲霉属10株试验菌中筛选得到一株产β-葡萄糖苷酶活性较高的黑曲霉FTA-008. 该菌株在适宜的培养条件下,β-葡萄糖苷酶的最高活性达2.89 U/mL,适宜产酶周期为3 d. 通过设计特异引物,以该菌株的总RNA为模板,采用RT-PCR技术获得大小为2 511 bp的cDNA片段,该序列与GenBank中AJ132386的β-葡萄糖苷酶序列相比,氨基酸序列同源性达96.8%. 经毕赤酵母表达,β-葡萄糖苷酶比活力达7.85 U/mg. 图3 表1 参13
Abstract:
From the 10 strains of Aspergillus, the strain A. niger FTA-008 was selected and it possessed high β-glucosidase activity (2.89 U/mL). The period of β-glucosidase production by this strain was 3 days. A set of primers were synthesized based on the sequence of β-glucosidase gene (bgl1) ( AJ132386) of A. niger. The cDNA fragment was amplified with RT-PCR using the total RNA of A. niger FTA-008 as template. The cloned gene fragment was 2 511 bp. Compared with sequence AJ132386, the identity of amino acid sequence was 96.8%. The cDNA was successfully expressed in Pichia pastoris and the maximum β-glucosidase activity (7.85 U/mg) was achieved in the BMMY medium containing 0.5% methanol and 240 r/min at pH 6.0 under 30 °C for 108 h. Fig 3, Tab 1, Ref 13

参考文献/References:

1 Wang JP (王建平), Chen XE (陈小娥). A survey study on cellulose. J Zhejiang Coll Fish (浙江水产学院院报), 1996, 15 (2): 140~144
2 Zhang HY (张鸿雁), Chen XS (陈锡时). 微生物纤维素酶分子生物学研究进展. Biotechnology (生物技术), 2003, 613 (3): 41~42
3 Wang TH (汪天虹), Wu J (吴静), Zou XX (邹小霞). Advances of molecular biology of Trichoderma reesei. Mycosystema (菌物系统), 2000, 19 (1): 147~152
4 Takashima S, Nakamura A, Hidaka M, Masaki H, Uozumi T. Molecular cloning and expression of the novel fungal beta-glucosidase genes from Humicola grisea and Trichoderma reesei. J Biochemistry, 1999, 125 (4): 728~736
5 Gu WM (顾卫民). The chatacteristic of β-glucosidase and its application in food industry. Jiangsu Food & Fermentation (江苏食品与发酵), 2003, 112 (1): 5~7
6 Brumbauer A, Reczey K. Beta-glucosidase production of two different Aspergillus strains. Acta Alimentaria, 1999, 28 (4): 361~370
7 Jiong H, Hisanori T, Hidehiko K. Cloning and functional expression of thermostable β-glucosidase gene from Thermoascus aurantiacus. Appl Microbiol Biotechnol (工业微生物), 2007, 73 (6): 1331~1339
8 Zhao LG (赵林果), Zhou TC (周潭澈), Meng P (孟鹏). Screening of β-glucosidase producing strain and analysis of cellulase composition. Ind Microbiol, 2007, 37 (5): 47~50
9 Ye JY (叶姜瑜). A new differential medium for cellulose decomposing microorganisms. Microbology (微生物学通报), 1997, 24 (4): 251~252
10 Montenecourt BS, Eveleigh DE. Preparation of mutants of Trichoderma reesei with enhanced cellulase production. Appl Environ Microbiol, 1977, 34 (6): 777~782
11 Thomas M, Wood K, Bhat M. Methods for measuring cellulose activities. Methods Enzymol, 1988, 160 (9): 87~111
12 Dellaporta SL, Wood J, Hicks JB. A plant DNA miniprepareation: Version Ⅱ. Plant Mol Biol Rep, 1983, 1 (4): 19
13 Michael EH, Mark FR, Charles E. Cellulase for commodity products from cellulosic biomass. Curr Opin Biotechnol, 1999, 10 (4): 358~364

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备注/Memo

备注/Memo:
*中国科学院百人计划项目、四川省青年科技基金项目(No. 08ZQ026-023)和中国科学院知识创新工程重大项目(No. KSCX1-YW-11B2)资助 Supported by the Program of 100 Distinguished Young Scientists of the Chinese Academy of Sciences, the Province Science Foundation for Young Scholars of Sichuan, China (No. 08ZQ026-023) and the Knowledge Innovation Program of the Chinese Academy of Sciences (No. KSCX1-YW-11B2)
**通讯作者 Corresponding author ( E-mail: chenjpro@163.com; wuzhl@cib.ac.cn)
更新日期/Last Update: 2009-07-03