Rapid extraction of phycobiliproteins from cultured cyanobacteria samples.
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 3h. The analysis of the extracted samples was carried out by capillary electrophoresis with laser-induced fluorescence detection.
Viskari
Pertti J
PJ
Department of Chemistry, Wake Forest University, Winston-Salem, NC 27109, USA.
Colyer
Christa L
CL
D002458:Cell Fractionation / Q000379:methods; D002478:Cells, Cultured; D000458:Cyanobacteria / Q000737:chemistry* / Q000166:cytology; D019075:Electrophoresis, Capillary; D045342:Light-Harvesting Protein Complexes / Q000302:isolation & purification*; D010833:Phytic Acid / Q000737:chemistry; D013050:Spectrometry, Fluorescence
D045342:Light-Harvesting Protein Complexes; D010833:Phytic Acid