Improving extracellular protein production in Escherichia coli by overexpressing D,D-carboxypeptidase to perturb peptidoglycan network synthesis and structure

doi:10.1007/s00253-018-9510-7

	Improving extracellular protein production in Escherichia coli by overexpressing D,D-carboxypeptidase to perturb peptidoglycan network synthesis and structure
	Yang, Haiquan 1; Hu, Jinyuan 1; Lu, Xiao 1; Wang, Fuxiang 1; Shen, Wei 1; Hu, Wei 2; Wang, Lingling 1; Chen, Xianzhong 1; Liu, Long 1
刊名	APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
	2019
卷号	103 页码:793-806
关键词	Extracellular protein production Overexpression D D-carboxypeptidase Peptidoglycan structure Membrane permeability Escherichia coli
ISSN号	0175-7598
DOI	10.1007/s00253-018-9510-7
英文摘要	Most recombinant proteins in Escherichia coli are not efficiently secreted to the extracellular space. Structural stabilisation of the cell wall is essential for extracellular protein production in E. coli, for which D,D-carboxypeptidases are essential. Herein, we perturbed the peptidoglycan structure of the E. coli cell wall by overexpressing D,D-carboxypeptidase genes dacA or dacB, and investigated the effect on extracellular protein production. Overexpression of dacA or dacB promoted the accumulation of intracellular soluble peptidoglycan, altered cell morphology (shape and size) and led to the formation of transparent globular structures in E. coli cells. Compared with controls (CK), extracellular production of recombinant green fluorescent protein (GFP) was increased by 1.7- and 2.3-fold upon overexpression of dacA and dacB, respectively. Similarly, extracellular production of recombinant amylase and -galactosidase was increased by 4.5- and 2.8-fold, respectively, upon overexpression of dacA, and by 11.9- and 2.5-fold, respectively, upon overexpression of dacB. Overexpression of dacA or dacB enhanced both the outer and inner membrane permeability of E. coli. This cell wall engineering strategy opens up a new direction for enhancing extracellular protein and chemical production in E. coli.
资助项目	National Natural Science Foundation of China[21406089] ; Natural Science Foundation of Jiangsu Province[BK20140152] ; Open Project Program of the Key Laboratory of Industrial Biotechnology, Ministry of Education, China[KLIB-KF201509] ; Open Project Program of the Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, China[KLCCB-KF201607] ; Open Project Program of the Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, China[KLCCB-KF201802] ; 111 Project[111-2-06] ; Postgraduate Education Research and Practice Project of Jiangnan University[YJSJG2017004]
WOS关键词	PENICILLIN-BINDING PROTEIN-5 ; HIGH-LEVEL EXPRESSION ; OUTER-MEMBRANE ; CELL-SHAPE ; RECOMBINANT PROTEINS ; ALPHA-GALACTOSIDASE ; BACILLUS-SUBTILIS ; L-FORM ; CARBOXYPEPTIDASE ; SECRETION
WOS研究方向	Biotechnology & Applied Microbiology
语种	英语
出版者	SPRINGER
WOS记录号	WOS:000457830000020
资助机构	National Natural Science Foundation of China ; Natural Science Foundation of Jiangsu Province ; Open Project Program of the Key Laboratory of Industrial Biotechnology, Ministry of Education, China ; Open Project Program of the Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, China ; 111 Project ; Postgraduate Education Research and Practice Project of Jiangnan University
内容类型	期刊论文
源URL	[http://119.78.100.186/handle/113462/66146]
专题	中国科学院近代物理研究所
作者单位	1.Jiangnan Univ, Minist Educ, Key Lab Carbohydrate Chem & Biotechnol, Wuxi 214122, Peoples R China 2.Chinese Acad Sci, Inst Modern Phys, Lanzhou 730000, Gansu, Peoples R China
推荐引用方式 GB/T 7714	Yang, Haiquan,Hu, Jinyuan,Lu, Xiao,et al. Improving extracellular protein production in Escherichia coli by overexpressing D,D-carboxypeptidase to perturb peptidoglycan network synthesis and structure[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2019,103:793-806.
APA	Yang, Haiquan.,Hu, Jinyuan.,Lu, Xiao.,Wang, Fuxiang.,Shen, Wei.,...&Liu, Long.(2019).Improving extracellular protein production in Escherichia coli by overexpressing D,D-carboxypeptidase to perturb peptidoglycan network synthesis and structure.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,103,793-806.
MLA	Yang, Haiquan,et al."Improving extracellular protein production in Escherichia coli by overexpressing D,D-carboxypeptidase to perturb peptidoglycan network synthesis and structure".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 103(2019):793-806.