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Cloning and functional characterization of the geranylgeranyl diphosphate synthase(GGPPS)from Elizabethkingia meningoseptica sp.F2
Yang, Qiang1,2; Zheng, Zhiming1; Zhao, Genhai1; Wang, Li1,2; Wang, Han1; Ni, Wenfeng1,2; Sun, Xiaowen1,2; Zhang, Mengxue1,2; Tang, Hengfang1,2; Wang, Peng1
2022
Source PublicationPROTEIN EXPRESSION AND PURIFICATION
ISSN1046-5928
Corresponding AuthorZheng, Zhiming(zhengzhiming2014@163.com) ; Wang, Peng(pengwang@ipp.ac.cn)
AbstractTo date, there is no functional characterization of EmGGPPS (from Elizabethkingia meningoseptica sp.F2) as enzymes catalyzing GGPP. In this research, maltose-binding protein (MBP), disulfide bond A (DbsA), disulfide bond C (DbsC), and two other small protein tags, GB1 (Protein G B1 domain) and ZZ (Protein A IgG ZZ repeat domain), were used as fusion partners to construct an EmGGPPS fusion expression system. The results indicated that the expression of MBP-EmGGPPS was higher than that of the other four fusion proteins in E. coli BL21 (DE3). Additionally, using EmGGPPS as a catalyst for the production of GGPP was verified using a color complementation assay in Escherichia coli. In parallel with it, the enzyme activity experiment in vitro showed that the EmGGPPS protein could produce GGPP, GPP and FPP. Finally, we successfully demonstrated MK-4 production in engineered E. coli by overexpression of EmGGPPS.
KeywordElizabethkingia meningoseptica Geranylgeranyl diphosphate synthase Soluble expression MK-4
DOI10.1016/j.pep.2021.105986
WOS KeywordESCHERICHIA-COLI ; ISOPRENOID BIOSYNTHESIS ; PROTEIN EXPRESSION ; BINDING DOMAIN ; VITAMIN-K ; PURIFICATION ; SYNTHASE ; PATHWAY ; YEAST
Indexed BySCI
Language英语
WOS Research AreaBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
WOS SubjectBiochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
WOS IDWOS:000709874400002
PublisherACADEMIC PRESS INC ELSEVIER SCIENCE
Citation statistics
Cited Times:1[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.hfcas.ac.cn:8080/handle/334002/125763
Collection中国科学院合肥物质科学研究院
Corresponding AuthorZheng, Zhiming; Wang, Peng
Affiliation1.Chinese Acad Sci, Inst Intelligent Machines, Hefei Inst Phys Sci, Hefei 230031, Peoples R China
2.Univ Sci & Technol China, Hefei 230026, Peoples R China
Recommended Citation
GB/T 7714
Yang, Qiang,Zheng, Zhiming,Zhao, Genhai,et al. Cloning and functional characterization of the geranylgeranyl diphosphate synthase(GGPPS)from Elizabethkingia meningoseptica sp.F2[J]. PROTEIN EXPRESSION AND PURIFICATION,2022,189.
APA Yang, Qiang.,Zheng, Zhiming.,Zhao, Genhai.,Wang, Li.,Wang, Han.,...&Wang, Peng.(2022).Cloning and functional characterization of the geranylgeranyl diphosphate synthase(GGPPS)from Elizabethkingia meningoseptica sp.F2.PROTEIN EXPRESSION AND PURIFICATION,189.
MLA Yang, Qiang,et al."Cloning and functional characterization of the geranylgeranyl diphosphate synthase(GGPPS)from Elizabethkingia meningoseptica sp.F2".PROTEIN EXPRESSION AND PURIFICATION 189(2022).
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