Institutional Repository of Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences
| Metabolic engineering of Escherichia coli for efficient biosynthesis of fluorescent phycobiliprotein | |
| Chen, Huaxin1,2,3; Jiang, Peng1,2,3 | |
| 2019-03-20 | |
| Source Publication | MICROBIAL CELL FACTORIES
 |
| ISSN | 1475-2859 |
| Volume | 18Pages:13 |
| Corresponding Author | Chen, Huaxin(chhx@qdio.ac.cn) |
| Abstract | BackgroundPhycobiliproteins (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. |
| Keyword | Phycobiliprotein Phycobilin Chromophorylation ratio Plasmid stability E. coli |
| DOI | 10.1186/s12934-019-1100-6 |
| Indexed By | SCI |
| Language | 英语 |
| Funding Project | development funds of science and technology of Shinan district, Qingdao[2016-3-010-ZH] ; National Natural Science Foundation of China[41276164] |
| WOS Research Area | Biotechnology & Applied Microbiology |
| WOS Subject | Biotechnology & Applied Microbiology |
| WOS ID | WOS:000462299800001 |
| Publisher | BMC |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://ir.qdio.ac.cn/handle/337002/154968 |
| Collection | 实验海洋生物学重点实验室 |
| Corresponding Author | Chen, Huaxin |
| Affiliation | 1.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 Affilication | Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences |
| Corresponding Author Affilication | Key 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. |
| Files in This Item: | There are no files associated with this item. | |||||
Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.
Edit Comment