title: Molecular Cloning of cpcU and Heterodimeric Bilin Lyase Activity Analysis of CpcU and CpcS for Attachment of Phycocyanobilin to Cys-82 on the β-Subunit of Phycocyanin in Arthrospira platensis FACHB314.
abstract: A new bilin lyase gene cpcU was cloned from Arthrospira platensis FACHB314 to study the assembly of the phycocyanin β-Subunit. Two recombinant plasmids, one contained the phycocyanobilin (PCB) producing genes (hoxI and pcyA), while the other contained the gene of the β-Subunit of phycobiliprotein (cpcB) and the lyase gene (cpcU, cpcS, or cpcU/S) were constructed and separately transferred into Escherichia coli in order to test the activities of relevant lyases for catalyzing PCB addition to CpcB during synthesizing fluorescent β-PC of A. platensis FACHB314. The fluorescence intensity examination showed that Cys-82 maybe the active site for the β-Subunit binding to PCBs and the attachment could be carried out by CpcU, CpcS, or co-expressed cpcU/S in A. platensis FACHB314.
authors.affiliation: Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China, Qingdao 266003, China. wufei0917@sina.cn.
authors.forename: Fei
authors.initials: F
authors.lastname: Wu
authors.affiliation: Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China, Qingdao 266003, China. xnzang@ouc.edu.cn.
authors.forename: Xiaonan
authors.initials: X
authors.lastname: Zang
authors.affiliation: Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China, Qingdao 266003, China. xczhang@ouc.edu.cn.
authors.forename: Xuecheng
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China, Qingdao 266003, China. ikaibei@163.com.
authors.forename: Ran
authors.initials: R
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China, Qingdao 266003, China. hxy764379760@163.com.
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Huang
authors.affiliation: Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China, Qingdao 266003, China. lu2014ouc@163.com.
authors.forename: Lulu
authors.initials: L
authors.lastname: Hou
authors.affiliation: Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China, Qingdao 266003, China. jiangmjp@163.com.
authors.forename: Minjie
authors.initials: M
authors.lastname: Jiang
authors.affiliation: Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China, Qingdao 266003, China. 17864275931@163.com.
authors.forename: Chang
authors.initials: C
authors.lastname: Liu
authors.affiliation: Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China, Qingdao 266003, China. pangchunhong2014@126.com.
authors.forename: Chunhong
authors.initials: C
authors.lastname: Pang
mesh_terms: D000595:Amino Acid Sequence; D001654:Bile Pigments / Q000378:metabolism*; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000145:classification / Q000201:enzymology* / Q000235:genetics*; D015870:Gene Expression; D008190:Lyases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D052980:Phycobilins / Q000378:metabolism*; D010798:Phycocyanin / Q000378:metabolism*; D010802:Phylogeny; D011485:Protein Binding; D055503:Protein Multimerization; D011994:Recombinant Proteins
chemical_list: D001654:Bile Pigments; D052980:Phycobilins; D011994:Recombinant Proteins; D010798:Phycocyanin; C011069:phycocyanobilin; D008190:Lyases
keywords: Arthrospira platensis FACHB314; CpcS; CpcU; fluorescence intensity; site-directed mutation