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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
ISSN1389-1723
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
DOI10.1016/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
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被引频次:12[WOS]   [WOS记录]     [WOS相关记录]
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
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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.
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