|本期目录/Table of Contents|

[1]张漫莉,耿新伟,王梦婷,等.纤维素酶EGA基因在枯草芽孢杆菌中的表达及其产物性质研究[J].浙江理工大学学报,2013,30(03):389-393.
 ZHANG Manli,GENG Xinwei,WANG Mengting,et al.Research on Expression of Cellulase EGA Gene in Bacillus Subtilis and Properties of Its Product[J].Journal of Zhejiang Sci-Tech University,2013,30(03):389-393.
点击复制

纤维素酶EGA基因在枯草芽孢杆菌中的表达及其产物性质研究()
分享到:

浙江理工大学学报[ISSN:1673-3851/CN:33-1338/TS]

卷:
第30卷
期数:
2013年03期
页码:
389-393
栏目:
生物与生命科学
出版日期:
2013-05-10

文章信息/Info

Title:
Research on Expression of Cellulase EGA Gene in Bacillus Subtilis and Properties of Its Product
文章编号:
1673-3851 (2013) 03-0389-05
作者:
张漫莉 耿新伟 王梦婷 蒋磊 赵辅昆 陈玮
浙江理工大学蛋白质组学与分子酶学实验室, 杭州 310018
Author(s):
ZHANG Manli GENG Xinwei WANG Mengting JIANG Lei ZHAO Fukun CHEN Wei
Laboratory of Proteomics and Molecular Enzymology, Zhejiang Sci-Tech University, Hangzhou 310018, China
关键词:
枯草芽孢杆菌 纤维素酶EGA 分泌 羧甲基纤维素钠
分类号:
Q786
文献标志码:
A
摘要:
枯草芽孢杆菌 Bacillus subtilis是一种高效的外源蛋白表达菌株,能将目的蛋白分泌到细胞外的培养基中。从福寿螺胃液共生菌株Bacillus sp. Strain AC 1基因组中克隆纤维素酶EGA的基因序列,利用基因工程技术构建纤维素酶EGA的重组枯草芽孢杆菌表达载体pAUsp ega 。该载体含有一个强启动子和一段信号肽,经淀粉诱导能表达外源蛋白。以枯草芽孢杆菌蛋白酶缺失型菌株WB700为宿主菌,表达得到的重组纤维素酶EGA以羧甲基纤维素钠(CMCNa)为底物得到的培养基上清酶活力达到1 120 U/L。该纤维素酶在pH 6.0表现最大水解活力,最适反应温度为60℃,在酸性条件下稳定性良好,具有较好的应用前景。

参考文献/References:

[1] 刘晓晶, 李田, 翟增强. 纤维素酶的研究现状及应用前景[J]. 安徽农业科学, 2011, 39(4): 1920-1921,1924.
[2] 康廷国. 中药鉴定学[M]. 北京: 中国中医药出版社, 2003: 36.
[3] 杨利平, 蔡水文, 罗玲, 等. 生物乙醇生产及纤维素酶的开发进展[J]. 西部资源, 2012(3): 132-134.
[4] Lynd L R, Weimer P J, Van Zyl W H, et al. Microbial cellulose utilization: fundamentals and biotechnology[J]. Microbiology and Molecular Biology Reviews, 2002, 66(3): 506-577.
[5] 刘萌, 战利, 红霞, 等. 纤维素酶及纤维素酶基因工程学研究进展[J]. 安徽农业科学, 2011, 39(16): 9515-9517.
[6] 黄妙容, 刘德武, 吴珍芳. 福寿螺多功能纤维素酶基因egx的克隆及其体外功能性表达[J].中国农业科学, 2011, 44(17): 3641-3648.
[7] Singh A, Hayashi K. Microbial cellulases: protein architecture, molecular properties and biosynthesis[J]. Advances in Applied Microbiology, 1995, 40:1-44.
[8] Eveleigh D E, Mandels M, Andreotti R, et al. Measurement of saccharifying cellulose[J]. Biotechnology for Biofuels, 2009, 2(1): 1-8.
[9] Webb E C. Enzyme nomenclature:a personal retrospective[J]. Federation of American Societies for Experimental Biology Journal, 1993, 7(12): 1192-1194.
[10] Li YanHong, Ding Ming, Wang Ji, et al. A novel thermoacidophilic eudoglucanase, BaEGA, from a new cellulose degrading bacterium,  Bacillus  sp. AC 1[J]. Apple Microbiol Bioechnol, 2006, 70(4): 430-436.
[11] Tjalsma H, Noback M A, Bron S, et al. Bacillus subtilis contains four closely related type I signal peptidases with overlapping substrate specificities: constitutive and temporally controlled expression of different sig gene[J]. J Biol Chem, 1997, 272(41): 25983-25992.
[12] Ogasawaia N. Systematic function analysis of Bacillus subtilis genes[J]. Res Microbiol, 2000, 151(2): 129-134.
[13] Doi R H, Wong S L, Kawamura. Potential use of bacillus subtilis for secretion and production of foreign proteins[J]. Trends Biotechnol, 1986, 4(9): 232-235.
[14] Chang S. Engineering for protein secretion in grampositive bacteria[J]. Methods Enzymol, 1987, 153: 507-516.
[15] Olmos S J, Contreras F R. Gentic system constructed to overproduce and secret proinsulin in Bacillus subtilis[J]. Appl Microbiol Biotechnol, 2003, 62(4): 369-373.
[16] Emilia M F, Duc L H, Isticato R, et al. Display of heterologous antigens on the Bacillus subtilis spore coat using CotC as a fusion partner[J]. Vaccine, 2004, 22(9/10): 1177-1187.
[17] BeraMaillet C, Arthaud L, Abad P, et al. Biochemical characterization of MIENG1, a family 5 endoglucanase secreted by the root knot nematode Meloidogyne incognita[J]. Eur J Biochem, 2000, 267(11): 3255-3263
[18] Nakashima K, Azuma J. Distribution and properties of endobeta1, 4glucanase from a lower termite, Coptotermes formosanus(Shiraki)[J]. Biosci Biotechnol Biochem, 2000, 64(7): 1500-1506.
[19] Xu B, Hellman U, Ersson B, et al. Purification, characterization and amino acid sequence analysis of a thermostable, low molecular mass endoβ1, 4glucanase from blue mussel, Mytilus edulis[J]. Eur J Biochem, 2000, 267(16): 4970-4977.
[20] Suzuki K I, Ojima T, Nishita K. Purification and cDNA cloning of a cellulase from abalone Haliotis discus hannai[J]. Eur J Biochem, 2003, 270 (4): 771-778.

相似文献/References:

[1]蒋红亮,张虹,赵辅昆,等.分子伴侣CsaA过表达及wprA蛋白酶的缺失对枯草芽孢杆菌分泌表达外源蛋白的影响[J].浙江理工大学学报,2011,28(02):260.

备注/Memo

备注/Memo:
收稿日期: 2012-11-06
基金项目: 浙江省大学生科技成果推广项目(2011R406051)
作者简介: 张漫莉(1989-),女,安徽阜阳人,硕士研究生,主要从事分子生物学和发酵工艺的研究。
通信作者: 陈玮,电子邮箱:cw@zstu.edu.cn
更新日期/Last Update: