Gewinnung und Anwendung von Phycoerythrin aus Porphyridium cruentum

Method for B-phycoerythrin purification from Porphyridium cruentum

“…Phycoerythrin (Fig. 2), obtained by the procedure described above, had an absorption spectrum typical for the pure B-PE (Gantt and Lipschultz, 1974;Roth et al, 1989) with peaks at 546 and 563 nm, and a shoulder at 498-500 nm. Its absorbance ratio As&i/A280 was 5 or higher, thus being a criterion for purity of B-PE preparations according to Gantt, (1969) When applied to gel filtration on Sepharose CL-GB, it was eluted as a single symmetric peak (Fig.…”

“…The latter is widely used as a fluorescent dye for mono-and multi-colour labeling in biochemical studies and clinical diagnostics that involve cell sorting, cell analyses and immunoassays (Kronick, 1986). It can be extracted from the algal cells with a phosphate buffer by heating the suspension at 30°C (Thepenier et al, 1987), by repeated freezing and thawing (Siegelman and Kycia, 1978), by using a French pressure cell (Leibo and Jones, 1964;Gantt et al, 1979) or by sonification (Jahn et al, 1984;Roth et al, 1989). In all cases the crude extract contains PBP and other water soluble components including high amounts of the polysaccharide typical for Porphyridium cruenfum.…”

“…Different combinations of high speed centrifugation, ammonium sulphate precipitation, chromatography on hydroxylapatite (O'hEocha, 1962;Roth et al, 1989;Sidler et al, 1989), hydrophobic chromatography (Leibo and Jones, 1964;Schoenleber et al, 1983;Roth et al, 1989) ion -exchange chromatography (Thepenier et al, 1987) gel filtration (Gantt, 1969;Schoenleber et al, 1983;Jahn et al, 1984) etc. were used for isolation and purification of B-PE from the crude extract.…”

Isolation and purification of phycoerythrin from red algaGracilaria verrucosa by expanded-bed-adsorption and ion-exchange chromatography

“…Means such as sonication [8], using a French pressure cell [9], heating extract suspension in a phosphate buffer at 40 ~ [10] and repeated freezing and thawing [11] and so on were used for ex-tracting algae, but the efficiency of all these methods is relatively low. After extracting algal material, different combinations of high speed centrifugation to remove cell debris and contaminants, ammonium sulfate precipitation, gel filtration [12], chromatography on hydroxylapatite [8,13], ion-exchange chromatography [10] and so on were used to isolate and purify phycobiliproteins. Compared to other organisms, marine algae always contain many more polysaccharides which are often liberated during homogenization of the algal cell and result in blockage of the chromatographic column [7], so large-scale isolation and purification of phycobiliproteins is difficult.…”