Metabolic engineering of Escherichia coli for efficient biosynthesis of fluorescent phycobiliprotein
文献类型：期刊论文
| 作者 | Chen, Huaxin1,2,3; Jiang, Peng1,2,3 |
| 刊名 | MICROBIAL CELL FACTORIES
 |
| 出版日期 | 2019-03-20 |
| 卷号 | 18页码:13 |
| 关键词 | Phycobiliprotein Phycobilin Chromophorylation ratio Plasmid stability E. coli |
| ISSN号 | 1475-2859 |
| DOI | 10.1186/s12934-019-1100-6 |
| 通讯作者 | Chen, Huaxin(chhx@qdio.ac.cn) |
| 英文摘要 | 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. |
| 资助项目 | development funds of science and technology of Shinan district, Qingdao[2016-3-010-ZH] ; National Natural Science Foundation of China[41276164] |
| WOS研究方向 | Biotechnology & Applied Microbiology |
| 语种 | 英语 |
| WOS记录号 | WOS:000462299800001 |
| 出版者 | BMC |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/154968]  |
| 专题 | 海洋研究所_实验海洋生物学重点实验室 |
| 通讯作者 | Chen, Huaxin |
| 作者单位 | 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 |
| 推荐引用方式 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. |
入库方式： OAI收割
来源：海洋研究所
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