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[1]卢潇,杨海泉,沈微,等.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):1114-1121.[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):1114-1121.[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:
1114-1121
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
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
Ministry of Education Key Laboratory of Carbohydrate Chemistry and Biotechnology, School of Biotechnology, Jiangnan University, Wuxi 214122, China
关键词:
DD-羧肽酶蛋白胞外生产大肠杆菌细胞形态
Keywords:
DD-carboxypeptidase protein extracellular production Escherichia coli cell morphology
分类号:
Q939.97 : Q786
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,但缺乏高效的蛋白胞外分泌机制,为提高其胞外蛋白生产水平,分析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. (图8 表2 参32)
Abstract:
Escherichia coli is one of the most widely used host strains for recombinant protein production, but it lacks efficient extracellular protein production capacity. It is of great significance to improve the extracellular protein production of E. coli. In this study, we analyzed the effect of overexpression of D,D-carboxypeptidase DacD on extracellular protein production by E. coli BL21 (DE3). The results showed that after overexpressing DacD in E. coli, the extracellular yield of recombinant green fluorescent protein (GFP) was 1.6 fold higher than that of the control strain (CK). In addition, the extracellular yield of recombinant amylase in E. coli BL21 was 2.6 fold higher than that in CK. Moreover, it was found that the overexpression of DacD promoted the accumulation of intracellular soluble peptidoglycan and that it enhanced inner membrane permeability of E. coli BL21. Overexpression of DacD resulted in a change in cell morphology, leading to the formation of transparent, globular structures in E. coli BL21 cells. In summary, overexpression of DacD provides a novel method for enhancing extracellular protein production in E. coli.

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更新日期/Last Update: 2018-10-25