Development of a plasmid-based, tunable, tolC-derived expression system for application in Cupriavidus necator H16

doi:10.1016/j.jbiotec.2018.03.007

CORC > 青岛生物能源与过程研究所 > 中国科学院青岛生物能源与过程研究所 > 材料生物技术研究中心

	Development of a plasmid-based, tunable, tolC-derived expression system for application in Cupriavidus necator H16
	Aboulnaga, Elhussiny A.1,2; Zou, Huibin 1,4; Selmer, Thorsten 3; Xian, Mo 1
刊名	JOURNAL OF BIOTECHNOLOGY
	2018-05-20
卷号	274 页码:15-27
关键词	Doxycycline Induced System Tolc-hybrid Promoter Glycerol Assimilation Mevalonate Production Cupriavidus Necator H16
ISSN号	0168-1656
DOI	10.1016/j.jbiotec.2018.03.007
文献子类	Article
英文摘要	Cupriavidus necator H16 gains increasing attention in microbial research and biotechnological application due to its diverse metabolic features. Here we present a tightly controlled gene expression system for C. necator including the pBBR1-vector that contains hybrid promoters originating from C. necator native tolC-promoter in combination with a synthetic tetO-operator. The expression of the reporter gene from these plasmids relies on the addition of the exogenous inducer doxycycline (dc). The novel expression system offers a combination of advantageous features as; (i) high and dose-dependent recombinant protein production, (ii) tight control with a high dynamic range (On/Off ratio), which makes it applicable for harmful pathways or for toxic protein production, (iii) comparable cheap inducer (doxycycline, dc), (iv) effective at low inducer concentration, that makes it useful for large scale application, (v) rapid, diffusion controlled induction, and (vi) the inducer does not interfere within the cell metabolism. As applications of the expression system in C. necator H16, the growth ability on glycerol was enhanced by constitutively expressing the E. coli glpk gene-encoding for glycerol kinase. Likewise, we used the system to overcome the expression toxicity of mevalonate pathway in C. necator H16. With this system, the mevalonate-genes were successfully introduced in the host and the recombinant strains could produce about 200 mg/l mevalonate.
WOS关键词	RALSTONIA-EUTROPHA H16 ; ESCHERICHIA-COLI ; ALCALIGENES-EUTROPHUS ; MULTIDRUG EFFLUX ; SOYBEAN OIL ; STRAIN H16 ; TRANSCRIPTIONAL REPRESSORS ; PROTEIN EXPRESSION ; GENE-EXPRESSION ; REGULATOR ACRR
WOS研究方向	Biotechnology & Applied Microbiology
语种	英语
出版者	ELSEVIER SCIENCE BV
WOS记录号	WOS:000430280000003
资助机构	Chinese Academy of Science (CAS President's International Fellowship Initiative, PIFI) ; Shandong Province Natural Science Foundation(ZR2015BM011 ; National Natural Science Foundation(21376255 ; 2016GSF121013) ; 21572242)
内容类型	期刊论文
源URL	[http://ir.qibebt.ac.cn/handle/337004/10811]
专题	青岛生物能源与过程研究所_材料生物技术研究中心
通讯作者	Aboulnaga, Elhussiny A.; Xian, Mo
作者单位	1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, CAS Key Lab Biobased Mat, Qingdao 266101, Peoples R China 2.Mansoura Univ, Fac Agr, Mansoura 35516, Egypt 3.Aachen Univ Appl Sci, Dept Chem & Biotechnol, Campus Juelich,Heinrich Mussmann Str 1, D-52428 Julich, Germany 4.Qingdao Univ Sci & Technol, Coll Chem Engn, Qingdao 266042, Peoples R China
推荐引用方式 GB/T 7714	Aboulnaga, Elhussiny A.,Zou, Huibin,Selmer, Thorsten,et al. Development of a plasmid-based, tunable, tolC-derived expression system for application in Cupriavidus necator H16[J]. JOURNAL OF BIOTECHNOLOGY,2018,274:15-27.
APA	Aboulnaga, Elhussiny A.,Zou, Huibin,Selmer, Thorsten,&Xian, Mo.(2018).Development of a plasmid-based, tunable, tolC-derived expression system for application in Cupriavidus necator H16.JOURNAL OF BIOTECHNOLOGY,274,15-27.
MLA	Aboulnaga, Elhussiny A.,et al."Development of a plasmid-based, tunable, tolC-derived expression system for application in Cupriavidus necator H16".JOURNAL OF BIOTECHNOLOGY 274(2018):15-27.