Establishing a Klebsiella pneumoniae-Based Cell-Free Protein Synthesis System

doi:10.3390/molecules27154684

	Establishing a Klebsiella pneumoniae-Based Cell-Free Protein Synthesis System
	Yang, Chen1 ; Yang, Miaomiao 2,3; Zhao, Wanhua 4; Ding, Yue 4; Wang, Yu 4; Li, Jian1
	2022-08
发表期刊	MOLECULES
EISSN	1420-3049
卷号	27 期号:15
发表状态	已发表
DOI	10.3390/molecules27154684
摘要	Cell-free protein synthesis (CFPS) systems are emerging as powerful platforms for in vitro protein production, which leads to the development of new CFPS systems for different applications. To expand the current CFPS toolkit, here we develop a novel CFPS system derived from a chassis microorganism Klebsiella pneumoniae, an important industrial host for heterologous protein expression and the production of many useful chemicals. First, we engineered the K. pneumoniae strain by deleting a capsule formation-associated wzy gene. This capsule-deficient strain enabled easy collection of the cell biomass for preparing cell extracts. Then, we optimized the procedure of cell extract preparation and the reaction conditions for CFPS. Finally, the optimized CFPS system was able to synthesize a reporter protein (superfolder green fluorescent protein, sfGFP) with a maximum yield of 253 +/- 15.79 mu g/mL. Looking forward, our K. pneumoniae-based CFPS system will not only expand the toolkit for protein synthesis, but also provide a new platform for constructing in vitro metabolic pathways for the synthesis of high-value chemicals.
关键词	Klebsiella pneumoniae cell-free protein synthesis genome editing cell-free synthetic biology
URL	查看原文
收录类别	SCI ; SCIE
语种	英语
资助项目	National Natural Science Foundation of China[31971348] ; Natural Science Foundation of Jiangxi Province["GJJ190239","20202BABL213040"]
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
WOS类目	Biochemistry & Molecular Biology ; Chemistry, Multidisciplinary
WOS记录号	WOS:000839689900001
出版者	MDPI
引用统计
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/219680
专题	物质科学与技术学院_硕士生物质科学与技术学院_PI研究组_李健组
通讯作者	Wang, Yu; Li, Jian
作者单位	1.ShanghaiTech Univ, Sch Phys Sci & Technol, Shanghai 201210, Peoples R China 2.Anhui Med Univ, Affiliated Hosp 4, Clin Pathol Ctr, Hefei 230012, Peoples R China 3.Univ Sci & Technol China, Dept Biol Phys, Hefei 230026, Peoples R China 4.Jiangxi Agr Univ, Coll Life Sci, Nanchang 330045, Jiangxi, Peoples R China
第一作者单位	物质科学与技术学院
通讯作者单位	物质科学与技术学院
第一作者的第一单位	物质科学与技术学院
推荐引用方式 GB/T 7714	Yang, Chen,Yang, Miaomiao,Zhao, Wanhua,et al. Establishing a Klebsiella pneumoniae-Based Cell-Free Protein Synthesis System[J]. MOLECULES,2022,27(15).
APA	Yang, Chen,Yang, Miaomiao,Zhao, Wanhua,Ding, Yue,Wang, Yu,&Li, Jian.(2022).Establishing a Klebsiella pneumoniae-Based Cell-Free Protein Synthesis System.MOLECULES,27(15).
MLA	Yang, Chen,et al."Establishing a Klebsiella pneumoniae-Based Cell-Free Protein Synthesis System".MOLECULES 27.15(2022).