海洋科学2017，Vol.41Issue(3)：68-74,7.DOI:10.11759/hykx20160512001
利用多功能裂合酶CpcE/F合成非天然重组藻胆蛋白
Biosynthesis of unnatural phycobiliproteins using universal bilin lyase CpcE/F
摘要
Abstract
Phycobilin lyase enzymes can attach phcobilins to their cognate apoproteins, which play an important role in the biosynthesis of phycobiliproteins. To verify the universal function of lyase cpcE/F, two recombinant plasmids: pCDFDuet-apcA-cpcE/F-hox1-pebS and pCDFDuet-pecA-cpcE/F-hox1-pebS were constructed using ge-netic engineering methods, and then two unnatural phycobiliproteins, ApcA-PEB and PecA-PEB, were successfully expressed in Escherichia coli, which indicated that cpcE/F could catalyze not only the ligation of PCB with differ-ent apoproteins, but also that of PEB and different apoproteins, and thus are universal bilin lyases. The absorption and fluorescence spectrum analysis showed that both unnatural proteins exhibit similar characteristic peaks to those of natural phycobiliproteins with some peaks of isomers and wavelength shifts, and their spectroscopic characteris-tics were mainly determined by the bilin type carried on them. Furthermore, different apoproteins could also regu-late the absorption spectrum and show important influences on the photostability of phycobilins. Our work will provide important information to better understand the biosynthetic mechanism of the phycocybiliproteins and production of unnatural phycobiliproteins with a better fluorescence quality.关键词
藻胆蛋白/生物合成/非天然/CpcE/F裂合酶Key words
phycocybiliproteins/biosynthesis/unnatural/CpcE/F lyase分类
生物科学引用本文复制引用
吝晓君,王祥法,葛保胜,秦松..利用多功能裂合酶CpcE/F合成非天然重组藻胆蛋白[J].海洋科学,2017,41(3):68-74,7.基金项目
国家高技术研究发展计划(863 计划)(No. 2014AA093505) （863 计划）
国家自然科学基金项目(No. 41176144) [Foundation: National High Technology Research and Development Pro-gram of China (863 project) (No. 2014AA093505) （No. 41176144）
national natural sci-ence fund project (No. 41176144)] （No. 41176144）
