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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
Source PublicationJOURNAL OF BIOTECHNOLOGY
ISSN0168-1656
Volume274Pages:15-27
AbstractCupriavidus 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.
SubtypeArticle
KeywordDoxycycline Induced System Tolc-hybrid Promoter Glycerol Assimilation Mevalonate Production Cupriavidus Necator H16
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
DOI10.1016/j.jbiotec.2018.03.007
WOS KeywordRALSTONIA-EUTROPHA H16 ; ESCHERICHIA-COLI ; ALCALIGENES-EUTROPHUS ; MULTIDRUG EFFLUX ; SOYBEAN OIL ; STRAIN H16 ; TRANSCRIPTIONAL REPRESSORS ; PROTEIN EXPRESSION ; GENE-EXPRESSION ; REGULATOR ACRR
Indexed BySCI
Language英语
WOS Research AreaBiotechnology & Applied Microbiology
Funding OrganizationChinese Academy of Science (CAS President's International Fellowship Initiative, PIFI) ; Shandong Province Natural Science Foundation(ZR2015BM011 ; National Natural Science Foundation(21376255 ; 2016GSF121013) ; 21572242)
WOS SubjectBiotechnology & Applied Microbiology
WOS IDWOS:000430280000003
PublisherELSEVIER SCIENCE BV
Citation statistics
Cited Times:1[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.qibebt.ac.cn/handle/337004/10812
Collection材料生物技术研究中心
Corresponding AuthorAboulnaga, Elhussiny A.; Xian, Mo
Affiliation1.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
Recommended Citation
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.
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