Rapid extraction of phycobiliproteins from cultured cyanobacteria samples

被引:87
作者
Viskari, PJ [1 ]
Colyer, CL [1 ]
机构
[1] Wake Forest Univ, Dept Chem, Winston Salem, NC 27109 USA
基金
美国国家科学基金会;
关键词
phycobiliproteins; Synechococcus; cyanobacteria; cell disruption;
D O I
10.1016/S0003-2697(03)00294-X
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Cyanobacteria, are a valuable and ubiquitous component of marine picophytoplankton that contribute significantly to total carbon biomass and primary productivity of the oceans. They contain water soluble, natively highly fluorescent proteins, phycobiliproteins, that can be considered ideal marker pigments for understanding the distribution and trophic dynamics of picoplankton populations. However, there is no standard protocol for extracting and quantitating these proteins from cyanobacterial cells. Ideally, the cells would be disrupted quickly and efficiently with complete extraction and recovery of the released proteins. For that purpose, we describe a method for extracting phycobiliproteins from a Synechococcus CCMP 833 cyanobacteria culture that utilizes 3% 3-[(3-cholamidopropyl)dimethyammonio]propanesulfonic acid (Chaps) 0.3% asolectin combined with nitrogen cavitation. Extraction efficiencies of greater than 85% were achieved by this method, which requires less than 3 h. The analysis of the extracted samples was carried out by capillary electrophoresis with laser-induced fluorescence detection. (C) 2003 Elsevier Science (USA). All rights reserved.
引用
收藏
页码:263 / 271
页数:9
相关论文
共 26 条
[11]  
JEFFREY SW, 1997, PHYTOPLANKTON PIGMEN, P152
[12]   CHROOCOCCOID CYANOBACTERIA IN THE SEA - UBIQUITOUS AND DIVERSE PHOTOTROPIC BIOMASS [J].
JOHNSON, PW ;
SIEBURTH, JM .
LIMNOLOGY AND OCEANOGRAPHY, 1979, 24 (05) :928-935
[13]  
KILPATRICK KA, 1985, THESIS TEXAS A M U, P12
[14]  
Lemberg R, 1949, HEMATIN COMPOUNDS BI
[15]  
Román RB, 2002, J BIOTECHNOL, V93, P73
[16]   EXTRACTION, IDENTIFICATION, AND QUANTITATION OF PHYCOBILIPROTEIN PIGMENTS FROM PHOTOTROPHIC PLANKTON [J].
STEWART, DE ;
FARMER, FH .
LIMNOLOGY AND OCEANOGRAPHY, 1984, 29 (02) :392-397
[17]   ADAPTATION OF SYNECHOCOCCUS INSITU DETERMINED BY VARIABILITY IN INTRACELLULAR PHYCOERYTHRIN-543 AT A COASTAL STATION OFF THE SOUTHERN CALIFORNIA COAST, USA [J].
VERNET, M ;
MITCHELL, BG ;
HOLMHANSEN, O .
MARINE ECOLOGY PROGRESS SERIES, 1990, 63 (01) :9-16
[18]   Separation and quantitation of phycobiliproteins using phytic acid in capillary electrophoresis with laser-induced fluorescence detection [J].
Viskari, PJ ;
Colyer, CL .
JOURNAL OF CHROMATOGRAPHY A, 2002, 972 (02) :269-276
[19]  
Viskari PJ, 2001, ELECTROPHORESIS, V22, P2327, DOI 10.1002/1522-2683(20017)22:11<2327::AID-ELPS2327>3.0.CO
[20]  
2-J