| Biosynthesis of a stable allophycocyanin beta subunit in metabolically engineered Escherichia coli | |
Chen, Huaxin1; Lin, Hanzhi1,2; Li, Fuchao1; Jiang, Peng1; Qin, Song3
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| 发表期刊 | JOURNAL OF BIOSCIENCE AND BIOENGINEERING
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| ISSN | 1389-1723 |
| 2013-05-01 | |
| 卷号 | 115期号:5页码:485-489 |
| 关键词 | Allophycocyanin Biosynthesis Escherichia Coli Fluorescence Thermostability |
| 产权排序 | [Chen, Huaxin; Lin, Hanzhi; Li, Fuchao; Jiang, Peng] Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China; [Lin, Hanzhi] Chinese Acad Sci, Grad Univ, Beijing 100039, Peoples R China; [Qin, Song] Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China |
| 通讯作者 | Jiang, P (reprint author), Room 335,Biol Bldg,7 Nanhai Rd, Qingdao 266071, Peoples R China. jiangpeng@qdio.ac.cn |
| 作者部门 | 海岸带生物学与生物资源利用所重点实验室 |
| 英文摘要 | Allophycocyanin (APC) is widely used as a fluorescent tag for fluorescence detection techniques. In this study, the alpha pcB gene from a thermophilic cyanobacterium strain was cloned and ligated into an expression vector to construct a pathway for the biosynthesis of an allophycocyanin beta subunit (holo-apcBT) in Escherichia coli. Isopropyl beta-D-1-thiogalactopyranoside induction successfully reconstituted holo-apcBT in E. coli. The recombinant holo-apcB showed spectroscopic properties similar to native APC. The stability and spectroscopic properties of this protein were then compared with another recombinant allophycocyanin beta subunit (holo-apcBM) whose apcB gene was cloned from mesophilic cyanobacterium. At high temperatures and during the course of storage, holo-apcBT was significantly more stable than holo-apcBM. In addition, holo-apcBT had an unexpectedly higher extinction coefficient and fluorescence quantum yield than holo-apcBM, suggesting that holo-apcBT would be a promising fluorescent tag and serve as a substitute for native APC. (c) 2012, The Society for Biotechnology, Japan. All rights reserved.; Allophycocyanin (APC) is widely used as a fluorescent tag for fluorescence detection techniques. In this study, the alpha pcB gene from a thermophilic cyanobacterium strain was cloned and ligated into an expression vector to construct a pathway for the biosynthesis of an allophycocyanin beta subunit (holo-apcBT) in Escherichia coli. Isopropyl beta-D-1-thiogalactopyranoside induction successfully reconstituted holo-apcBT in E. coli. The recombinant holo-apcB showed spectroscopic properties similar to native APC. The stability and spectroscopic properties of this protein were then compared with another recombinant allophycocyanin beta subunit (holo-apcBM) whose apcB gene was cloned from mesophilic cyanobacterium. At high temperatures and during the course of storage, holo-apcBT was significantly more stable than holo-apcBM. In addition, holo-apcBT had an unexpectedly higher extinction coefficient and fluorescence quantum yield than holo-apcBM, suggesting that holo-apcBT would be a promising fluorescent tag and serve as a substitute for native APC. (c) 2012, The Society for Biotechnology, Japan. All rights reserved. |
| 文章类型 | Article |
| 资助机构 | Natural Science Foundation of Shandong Province [2009ZRB02542]; Foundation of the Key Laboratory of Marine Bioactive Substance and Modern Analytical Techniques, State Oceanic Administration [MBSMAT-2010-03]; Nonprofit Research Project for the State Oceanic Administration [200905021-3, 201205027-2] |
| 收录类别 | SCI |
| 语种 | 英语 |
| 关键词[WOS] | SYNECHOCOCCUS SP PCC-7002 ; C-PHYCOCYANIN ; CYANOBACTERIAL PHYCOBILIPROTEINS ; CPCS-I ; PROTEIN ; LYASE ; BIOGENESIS ; STABILITY ; PEPTIDES ; TRIMER |
| 研究领域[WOS] | Biotechnology & Applied Microbiology ; Food Science & Technology |
| WOS记录号 | WOS:000320208600004 |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.yic.ac.cn/handle/133337/6412 |
| 专题 | 海岸带生物学与生物资源利用重点实验室_海岸带生物学与生物资源保护实验室 |
| 作者单位 | 1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 100039, Peoples R China 3.Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China |
| 推荐引用方式 GB/T 7714 | Chen, Huaxin,Lin, Hanzhi,Li, Fuchao,et al. Biosynthesis of a stable allophycocyanin beta subunit in metabolically engineered Escherichia coli[J]. JOURNAL OF BIOSCIENCE AND BIOENGINEERING,2013,115(5):485-489. |
| APA | Chen, Huaxin,Lin, Hanzhi,Li, Fuchao,Jiang, Peng,&Qin, Song.(2013).Biosynthesis of a stable allophycocyanin beta subunit in metabolically engineered Escherichia coli.JOURNAL OF BIOSCIENCE AND BIOENGINEERING,115(5),485-489. |
| MLA | Chen, Huaxin,et al."Biosynthesis of a stable allophycocyanin beta subunit in metabolically engineered Escherichia coli".JOURNAL OF BIOSCIENCE AND BIOENGINEERING 115.5(2013):485-489. |
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