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Chromophore attachment to fusion protein of streptavidin and recombinant allophycocyanin alpha subunit
Wu, Jing1,2,3; Chen, Huaxin1,2; Jiang, Peng1,2
2018
Source PublicationBIOENGINEERED
ISSN2165-5979
Volume9Issue:1Pages:108-115
Corresponding AuthorChen, Huaxin(chenhx001@126.com) ; Jiang, Peng(Jiangpeng@qdio.ac.cn)
AbstractThe fusion protein (SLA) of streptavidin and allophycocyanin subunit (holo-ApcA) was biosynthesized in Escherichia coli by a dual plasmid system. The recombinant SLA, purified by affinity chromatography, showed spectral properties similar to natural allophycocyanin subunit (ApcA). Spectral and Zinc staining analysis indicated that the recombinant SLA covalently bound phycocyanobilin (PCB). To improve chromophorylation rate of recombinant SLA, an in vitro chromophore attachment reaction system was established, which contained partially chromophylated SLA, PCB and lyase CpcS. Spectral analysis showed that PCB bound to the recombinant SLA rapidly during the reaction. The chromophorylation rate of SLA was improved from 21.1% to 86.5%. Immunofluorescence assay showed that SLA with high chromophorylation rate had higher detection signal. Thus, in vitro chromophore attachment is an effective way to improve the chromophorylation rate of recombinant phycobiliprotein.
KeywordAllophycocyanin Chromophore attachment Chromophorylation rate Immunofluorescence assay Phycobiliprotein
DOI10.1080/21655979.2017.1321282
Indexed BySCI
Language英语
Funding ProjectNational Natural Science Foundation[41276164] ; National 863 Program[2014AA093505] ; National 863 Program[2014AA093501] ; development funds of science and technology of Shinan district[2016-3-010-ZH] ; Scientific and Technological Innovation Project - Qingdao National Laboratory for Marine Science and Technology[2015ASKJ02]
WOS Research AreaBiotechnology & Applied Microbiology
WOS SubjectBiotechnology & Applied Microbiology
WOS IDWOS:000425053900001
PublisherTAYLOR & FRANCIS INC
Citation statistics
Cited Times:1[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/158039
Collection实验海洋生物学重点实验室
Corresponding AuthorChen, Huaxin; Jiang, Peng
Affiliation1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China
2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao, Peoples R China
3.Univ Chinese Acad Sci, Beijing, Peoples R China
First Author AffilicationKey Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences
Corresponding Author AffilicationKey Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences
Recommended Citation
GB/T 7714
Wu, Jing,Chen, Huaxin,Jiang, Peng. Chromophore attachment to fusion protein of streptavidin and recombinant allophycocyanin alpha subunit[J]. BIOENGINEERED,2018,9(1):108-115.
APA Wu, Jing,Chen, Huaxin,&Jiang, Peng.(2018).Chromophore attachment to fusion protein of streptavidin and recombinant allophycocyanin alpha subunit.BIOENGINEERED,9(1),108-115.
MLA Wu, Jing,et al."Chromophore attachment to fusion protein of streptavidin and recombinant allophycocyanin alpha subunit".BIOENGINEERED 9.1(2018):108-115.
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