Purification of R-phycoerythrin from Porphyra haitanensis (Bangiales, Rhodophyta) using expanded-bed absorption

doi:10.1111/j.1529-8817.2007.00401.x

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	Purification of R-phycoerythrin from Porphyra haitanensis (Bangiales, Rhodophyta) using expanded-bed absorption
	Niu, Jian-Feng ; Wang, Guang-Ce ; Zhou, Bai-Cheng ; Lin, Xiang-Zhi ; Chen, Chang-Sheng ; Wang, GC, Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China
	2007-12-01
发表期刊	JOURNAL OF PHYCOLOGY
ISSN	0022-3646
卷号	43 期号:6 页码:1339-1347
文章类型	Article
摘要	R-phycoerythrin (R-PE) was purified from leafy gametophyte of Porphyra haitanensis T. J. Chang et B. F. Zheng (Bangiales, Rhodophyta) by a simple, scaleable procedure. Initially, phycobiliproteins were extracted by repeated freeze-thaw cycles, resulting in release from the algal cells by osmotic shock. Next, R-PE was recovered by applying the crude extract with a high concentration of (NH4)(2)SO4 salt directly to the expanded-bed columns loaded with phenyl-sepharose. An expanded-bed volume twice the settled-bed volume was maintained; then low (NH4)(2)SO4 concentration was used to develop the column. After two rounds of hydrophobic interaction chromatography (HIC), R-PE was purified by anion-exchange column. The method was also successful with free-living conchocelis of P. haitanensis. The purified R-PE was identified with electrophoresis, and absorption and fluorescence emission spectroscopy. The results were in agreement with those previously reported. The yield with a spectroscopic purity (OD565/OD280) higher than 3.2 (the ratio of A(565)/A(620) <= 0.02) was 1.4 mg . g(-1) of leafy gametophyte of P. haitanensis. For the free-living conchocelis of P. haitanensis extract, R-PE could be purified successfully with only one round of HIC. The yield with a spectroscopic purity (OD565/OD280) higher than 3.2 (the ratio of A(565)/A(620) <= 0.02) was 5.0 mg . g(-1) of free-living conchocelis of P. haitanensis. The method described here is a scaleable technology that allows a large quantity of R-PE to be recovered from the unclarified P. haitanensis crude extract. It is also a high protein recovery technology, reducing both processing costs and times, which enhances the value of this endemic Porphyra of China.; R-phycoerythrin (R-PE) was purified from leafy gametophyte of Porphyra haitanensis T. J. Chang et B. F. Zheng (Bangiales, Rhodophyta) by a simple, scaleable procedure. Initially, phycobiliproteins were extracted by repeated freeze-thaw cycles, resulting in release from the algal cells by osmotic shock. Next, R-PE was recovered by applying the crude extract with a high concentration of (NH4)(2)SO4 salt directly to the expanded-bed columns loaded with phenyl-sepharose. An expanded-bed volume twice the settled-bed volume was maintained; then low (NH4)(2)SO4 concentration was used to develop the column. After two rounds of hydrophobic interaction chromatography (HIC), R-PE was purified by anion-exchange column. The method was also successful with free-living conchocelis of P. haitanensis. The purified R-PE was identified with electrophoresis, and absorption and fluorescence emission spectroscopy. The results were in agreement with those previously reported. The yield with a spectroscopic purity (OD565/OD280) higher than 3.2 (the ratio of A(565)/A(620) <= 0.02) was 1.4 mg . g(-1) of leafy gametophyte of P. haitanensis. For the free-living conchocelis of P. haitanensis extract, R-PE could be purified successfully with only one round of HIC. The yield with a spectroscopic purity (OD565/OD280) higher than 3.2 (the ratio of A(565)/A(620) <= 0.02) was 5.0 mg . g(-1) of free-living conchocelis of P. haitanensis. The method described here is a scaleable technology that allows a large quantity of R-PE to be recovered from the unclarified P. haitanensis crude extract. It is also a high protein recovery technology, reducing both processing costs and times, which enhances the value of this endemic Porphyra of China.
关键词	Anion-exchange Chromatography Hydrophobic Interaction Chromatography Porphyra Haitanensis R-phycoerythrin Streamline Column
学科领域	Plant Sciences ; Marine & Freshwater Biology
DOI	10.1111/j.1529-8817.2007.00401.x
URL	查看原文
收录类别	SCI
语种	英语
WOS记录号	WOS:000251416100023
引用统计	被引频次：31[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/337002/1653
专题	实验海洋生物学重点实验室
通讯作者	Wang, GC, Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China
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 Sch Sci, Beijing 100039, Peoples R China 3.Tianjin Univ Sci & Technol, Coll Marine Sci & Engn, Tianjin 300457, Peoples R China 4.Inst Oceanog 3, State Ocean Adm Peoples Republ China, Xiamen 361005, Peoples R China 5.Jimei Univ, Xiamen 361021, Peoples R China
推荐引用方式 GB/T 7714	Niu, Jian-Feng,Wang, Guang-Ce,Zhou, Bai-Cheng,et al. Purification of R-phycoerythrin from Porphyra haitanensis (Bangiales, Rhodophyta) using expanded-bed absorption[J]. JOURNAL OF PHYCOLOGY,2007,43(6):1339-1347.
APA	Niu, Jian-Feng,Wang, Guang-Ce,Zhou, Bai-Cheng,Lin, Xiang-Zhi,Chen, Chang-Sheng,&Wang, GC, Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China.(2007).Purification of R-phycoerythrin from Porphyra haitanensis (Bangiales, Rhodophyta) using expanded-bed absorption.JOURNAL OF PHYCOLOGY,43(6),1339-1347.
MLA	Niu, Jian-Feng,et al."Purification of R-phycoerythrin from Porphyra haitanensis (Bangiales, Rhodophyta) using expanded-bed absorption".JOURNAL OF PHYCOLOGY 43.6(2007):1339-1347.

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