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[1]卢 潇 杨海泉** 沈 微** 许 菲.DacD过量表达对大肠杆菌胞外蛋白生产的影响*[J].应用与环境生物学报,2018,24(05):1-13.[doi:10.19675/j.cnki.1006-687x.2017.10029]
 LU Xiao,YANG Haiquan **,SHEN Wei** & XU Fei.Effect of overexpressing DacD on extracellular protein production in Escherichia coli*[J].Chinese Journal of Applied & Environmental Biology,2018,24(05):1-13.[doi:10.19675/j.cnki.1006-687x.2017.10029]
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DacD过量表达对大肠杆菌胞外蛋白生产的影响*()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
24卷
期数:
2018年05期
页码:
1-13
栏目:
研究论文
出版日期:
2018-10-25

文章信息/Info

Title:
Effect of overexpressing DacD on extracellular protein production in Escherichia coli*
文章编号:
201710029
作者:
卢 潇 杨海泉** 沈 微** 许 菲
江南大学生物工程学院糖化学与生物技术教育部重点实验室 无锡 214122
Author(s):
LU Xiao YANG Haiquan ** SHEN Wei** & XU Fei
Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education , Jiangnan University , Wuxi 214122, China
关键词:
D D -羧肽酶提高蛋白胞外生产大肠杆菌细胞形态
Keywords:
DD-carboxypeptidase improve protein extracellular production Escherichia coli cell morphology
DOI:
10.19675/j.cnki.1006-687x.2017.10029
摘要:
大肠杆菌(Escherichia coli)是广泛用于重组蛋白表达的宿主之一。但E. coli 缺乏高效的蛋白胞外分泌机制,研究如何提高E. coli 胞外蛋白生产水平具有重要意义。研究中分析了D,D-羧肽酶DacD过量表达对E. coli 胞外蛋白生产水平的影响。结果表明:与对照株相比,过量表达DacD时重组E. coli BL21 (DE3)的重组绿色荧光蛋白(GFP)的胞外生产水平提高了1.6倍。同时,当过量表达DacD时,E. coli BL21 的重组淀粉酶胞外生产水平较对照株提高了2.6倍。进一步研究发现,DacD的过量表达可促进E. coli BL21 细胞内可溶性肽聚糖的积累。过量表达DacD也增强了E. coli BL21 细胞内膜的通透性。DacD过量表达导致了E. coli BL21 细胞形态的变化,重组菌株细胞两端形成了透明的球状囊泡结构。因此,通过过量DacD提高E. coli 胞外蛋白生产水平是可行的,结果可为促进E. coli 蛋白胞外生产提供了新的研究思路。(图8 表2 参35)
Abstract:
Escherichia coli is one of the most widely used host strains for recombinant protein production, but E. coli lacks high efficient extracellular protein production capacity. It is of great significance to study how to improve the extracellular protein production in E. coli. In this work, we analyzed that the effect of overexpression of D,D-carboxypeptidase DacD on the extracellular protein production in Escherichia coli. Results: after expressing DacD, the extracellular yield of recombinant green fluorescent protein (GFP) in E. coli BL21 (DE3) was improved by 1.6-fold of that of the control strain (CK) . Meanwhile, the extracellular yield of recombinant amylase in E. coli BL21was increased by 2.6-fold of that of CK. It was also found that overexpression of DacD promoted the accumulation of intracellular soluble peptidoglycan of E. coli BL21. Overexpressing DacD enhanced inner membrane permeability of E. coli BL21 . Overexpression of DacD resulted in change of cell m orphology and formed transparent globular structures in E. coli BL21 cells. In summary, level of extracellular protein production in E. coli was successfully enhanced by overexpressing DacD, which provide a new method to extracellular production of protein s in E. coli.

备注/Memo

备注/Memo:
收稿日期: 2017-10-26 接受日期: 2018-01-24
*国家自然科学基金项目(21406089)、江苏省自然科学基金(BK20140152)、江南大学研究生教育教学研究与实践课题(YJSJG2017004)资助?
**通讯作者(E-mail: haiquanyang@jiangnan.edu.cn; shenwei_micro@163.com)
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更新日期/Last Update: 2018-04-11