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.

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.
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DOI:
10.3390/molecules21030357
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发表时间:
2016-03-16
期刊:
Molecules (Basel, Switzerland)
影响因子:
--
通讯作者:
Pang C
Pang C
中科院分区:
其他
文献类型:
--
作者:
Wu F;Zang X;Zhang X;Zhang R;Huang X;Hou L;Jiang M;Liu C;Pang C

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从钝顶螺旋藻FACHB314中克隆了一个新的胆碱裂解酶基因Cpcu,用于研究藻蓝蛋白β亚基的组装。构建了含有藻蓝胆素产生基因(hoxI和pcyA)、藻胆蛋白β亚基基因(Cpc)和裂解酶基因(cpcU、cpS或cpcu/S)的重组表达载体,并将它们分别转化到大肠杆菌中,以检测相关裂解酶在钝顶螺旋藻FACHB314合成荧光β-PC过程中催化ccb加成的活性。荧光强度检测表明,Cys-82可能是β亚基与多氯联苯结合的活性部位,在钝顶螺旋藻FACHB314中,Cys-82可能通过CpCU、CpC或共表达CpCu/S进行结合。
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.
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