Improving extracellular protein production in Escherichia coli by overexpressing D,D-carboxypeptidase to perturb peptidoglycan network synthesis and structure
文献类型：期刊论文
| 作者 | Yang, Haiquan1; Hu, Jinyuan1; Lu, Xiao1; Wang, Fuxiang1; Shen, Wei1; Hu, Wei2; Wang, Lingling1; Chen, Xianzhong1; Liu, Long1 |
| 刊名 | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
 |
| 出版日期 | 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. |
| 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 |
| 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研究方向 | Biotechnology & Applied Microbiology |
| 语种 | 英语 |
| WOS记录号 | WOS:000457830000020 |
| 出版者 | SPRINGER |
| 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 |
| 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. |
入库方式： OAI收割
来源：近代物理研究所
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