Institutional Repository of Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences
Biosynthesis of a stable allophycocyanin beta subunit in metabolically engineered Escherichia coli | |
Chen, Huaxin1; Lin, Hanzhi1,2; Li, Fuchao1; Jiang, Peng1; Qin, Song3; Jiang, P | |
2013-05-01 | |
发表期刊 | JOURNAL OF BIOSCIENCE AND BIOENGINEERING |
ISSN | 1389-1723 |
卷号 | 115期号:5页码:485-489 |
文章类型 | Article |
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. |
关键词 | Allophycocyanin Biosynthesis Escherichia Coli Fluorescence Thermostability |
学科领域 | Biotechnology & Applied Microbiology ; Food Science & Technology |
DOI | 10.1016/j.jbiosc.2012.11.008 |
URL | 查看原文 |
收录类别 | SCI |
语种 | 英语 |
WOS研究方向 | Biotechnology & Applied Microbiology ; Food Science & Technology |
WOS类目 | Biotechnology & Applied Microbiology ; Food Science & Technology |
WOS记录号 | WOS:000320208600004 |
WOS关键词 | SYNECHOCOCCUS SP PCC-7002 ; C-PHYCOCYANIN ; CYANOBACTERIAL PHYCOBILIPROTEINS ; CPCS-I ; PROTEIN ; LYASE ; BIOGENESIS ; STABILITY ; PEPTIDES ; TRIMER |
WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
引用统计 | |
文献类型 | 期刊论文 |
条目标识符 | http://ir.qdio.ac.cn/handle/337002/16534 |
专题 | 实验海洋生物学重点实验室 |
通讯作者 | Jiang, P |
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 |
第一作者单位 | 实验海洋生物学重点实验室 |
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,&Jiang, P.(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. |
条目包含的文件 | ||||||
文件名称/大小 | 文献类型 | 版本类型 | 开放类型 | 使用许可 | ||
Biosynthesis of a st(539KB) | 限制开放 | CC BY-NC-SA | 浏览 |
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