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Development of a plasmid-based, tunable, tolC-derived expression system for application in Cupriavidus necator H16
Aboulnaga, Elhussiny A.1,2; Zou, Huibin1,4; Selmer, Thorsten3; Xian, Mo1
2018-05-20
发表期刊JOURNAL OF BIOTECHNOLOGY
ISSN0168-1656
卷号274页码:15-27
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.
文章类型Article
关键词Doxycycline Induced System Tolc-hybrid Promoter Glycerol Assimilation Mevalonate Production Cupriavidus Necator H16
WOS标题词Science & Technology ; Life Sciences & Biomedicine
DOI10.1016/j.jbiotec.2018.03.007
关键词[WOS]RALSTONIA-EUTROPHA H16 ; ESCHERICHIA-COLI ; ALCALIGENES-EUTROPHUS ; MULTIDRUG EFFLUX ; SOYBEAN OIL ; STRAIN H16 ; TRANSCRIPTIONAL REPRESSORS ; PROTEIN EXPRESSION ; GENE-EXPRESSION ; REGULATOR ACRR
收录类别SCI
语种英语
WOS研究方向Biotechnology & Applied Microbiology
项目资助者Chinese Academy of Science (CAS President's International Fellowship Initiative, PIFI) ; Shandong Province Natural Science Foundation(ZR2015BM011 ; National Natural Science Foundation(21376255 ; 2016GSF121013) ; 21572242)
WOS类目Biotechnology & Applied Microbiology
WOS记录号WOS:000430280000003
出版者ELSEVIER SCIENCE BV
引用统计
文献类型期刊论文
条目标识符http://ir.qibebt.ac.cn/handle/337004/10812
专题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
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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.
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