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Metabolic engineering of Escherichia coli for efficient biosynthesis of fluorescent phycobiliprotein
Chen, Huaxin1,2,3; Jiang, Peng1,2,3
2019-03-20
Source PublicationMICROBIAL CELL FACTORIES
ISSN1475-2859
Volume18Pages:13
Corresponding AuthorChen, Huaxin(chhx@qdio.ac.cn)
AbstractBackgroundPhycobiliproteins (PBPs) are light-harvesting protein found in cyanobacteria, red algae and the cryptomonads. They have been widely used as fluorescent labels in cytometry and immunofluorescence analysis. A number of PBPs has been produced in metabolically engineered Escherichia coli. However, the recombinant PBPs are incompletely chromophorylated, and the underlying mechanisms are not clear.Results and discussionIn this work, a pathway for SLA-PEB [a fusion protein of streptavidin and allophycocyanin that covalently binds phycoerythrobilin (PEB)] biosynthesis in E. coli was constructed using a single-expression plasmid strategy. Compared with a previous E. coli strain transformed with dual plasmids, the E. coli strain transformed with a single plasmid showed increased plasmid stability and produced SLA-PEB with a higher chromophorylation ratio. To achieve full chromophorylation of SLA-PEB, directed evolution was employed to improve the catalytic performance of lyase CpcS. In addition, the catalytic abilities of heme oxygenases from different cyanobacteria were investigated based on biliverdin IX and PEB accumulation. Upregulation of the heme biosynthetic pathway genes was also carried out to increase heme availability and PEB biosynthesis in E. coli. Fed-batch fermentation was conducted for the strain V5ALD, which produced recombinant SLA-PEB with a chromophorylation ratio of 96.7%.ConclusionIn addition to reporting the highest chromophorylation ratio of recombinant PBPs to date, this work demonstrated strategies for improving the chromophorylation of recombinant protein, especially biliprotein with heme, or its derivatives as a prosthetic group.
KeywordPhycobiliprotein Phycobilin Chromophorylation ratio Plasmid stability E. coli
DOI10.1186/s12934-019-1100-6
Indexed BySCI
Language英语
Funding Projectdevelopment funds of science and technology of Shinan district, Qingdao[2016-3-010-ZH] ; National Natural Science Foundation of China[41276164]
WOS Research AreaBiotechnology & Applied Microbiology
WOS SubjectBiotechnology & Applied Microbiology
WOS IDWOS:000462299800001
PublisherBMC
Citation statistics
Document Type期刊论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/154968
Collection实验海洋生物学重点实验室
Corresponding AuthorChen, Huaxin
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 266071, Peoples R China
3.Chinese Acad Sci, Ctr Ocean Mega Sci, Qingdao 266071, 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
Chen, Huaxin,Jiang, Peng. Metabolic engineering of Escherichia coli for efficient biosynthesis of fluorescent phycobiliprotein[J]. MICROBIAL CELL FACTORIES,2019,18:13.
APA Chen, Huaxin,&Jiang, Peng.(2019).Metabolic engineering of Escherichia coli for efficient biosynthesis of fluorescent phycobiliprotein.MICROBIAL CELL FACTORIES,18,13.
MLA Chen, Huaxin,et al."Metabolic engineering of Escherichia coli for efficient biosynthesis of fluorescent phycobiliprotein".MICROBIAL CELL FACTORIES 18(2019):13.
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