Biosynthesis of a Phycocyanin Beta Subunit with Two Noncognate Chromophores in Escherichia coli

doi:10.1007/s12010-019-03219-3

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	Biosynthesis of a Phycocyanin Beta Subunit with Two Noncognate Chromophores in Escherichia coli
	Chen, Huaxin 1,2,3; Zheng, Caiyun 4; Jiang, Peng 1,2,3; Ji, Gengsheng 4
刊名	APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
	2020-06-01
卷号	191 期号:2 页码:763-771
关键词	Phycobiliprotein Phycocyanin Lyase Fluorescence Chromophore E coli
ISSN号	0273-2289
DOI	10.1007/s12010-019-03219-3
通讯作者	Chen, Huaxin(chhx@qdio.ac.cn)
英文摘要	Recombinant phycobiliprotein can be used as fluorescent label in immunofluorescence assay. In this study, pathway for phycocyanin beta subunit (CpcB) carrying noncognate chromophore phycoerythrobilin (PEB) and phycourobilin (PUB) was constructed in Escherichia coli. Lyase CpcS and CpcT could catalyze attachment of PEB to Cys84-CpcB and Cys155-CpcB, respectively. However, PEB was attached only to Cys84-CpcB when both CpcS and CpcT were present in E. coli. A dual plasmid expression system was used to control the expression of lyases and the attachment order of PEB to CpcB. The production of PEB-Cys155-CpcB was achieved by L-arabinose-induced expression of CpcS, CpcB, Ho1, and PebS, and then the attachment of PEB to Cys84-CpcB was achieved by IPTG-induced expression of CpcS. The doubly chromophorylated CpcB absorbed light maximally at 497.5 nm and 557.0 nm and fluoresced maximally at 507.5 nm and 566.5 nm. An amount of light energy absorbed by PUB-Cys155-CpcB is transferred to PEB-Cys84-CpcB in doubly chromophorylated CpcB, conferring a large stokes shift of 69 nm for this fluorescent protein. There are interactions between chromophores of CpcB which possibly together with the help of lyases lead to isomerization of PEB-Cys155-CpcB to PUB-Cys155-CpcB.
WOS研究方向	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
语种	英语
出版者	SPRINGER
WOS记录号	WOS:000541559900021
内容类型	期刊论文
源URL	[http://ir.qdio.ac.cn/handle/337002/167804]
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.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China 3.Chinese Acad Sci, Ctr Ocean Mega Sci, Qingdao 266071, Peoples R China 4.Jiangsu Univ Sci & Technol, Coll Biotechnol Sericultural Res Inst, Zhenjiang, Jiangsu, Peoples R China
推荐引用方式 GB/T 7714	Chen, Huaxin,Zheng, Caiyun,Jiang, Peng,et al. Biosynthesis of a Phycocyanin Beta Subunit with Two Noncognate Chromophores in Escherichia coli[J]. APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY,2020,191(2):763-771.
APA	Chen, Huaxin,Zheng, Caiyun,Jiang, Peng,&Ji, Gengsheng.(2020).Biosynthesis of a Phycocyanin Beta Subunit with Two Noncognate Chromophores in Escherichia coli.APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY,191(2),763-771.
MLA	Chen, Huaxin,et al."Biosynthesis of a Phycocyanin Beta Subunit with Two Noncognate Chromophores in Escherichia coli".APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY 191.2(2020):763-771.