Improved procedure for separation and purification of Arthronema africanum phycobiliproteins

Abstract

A rapid, inexpensive and reliable procedure for separation and purification of C-phycocyanin (C-PC) and allophycocyanin (APC) from Arthronema africanum based on a previously described rivanol-sulfate method for C-PC purification was developed. Exclusion of NaCl from the extraction buffer resulted in complete separation of APC and C-PC, two-fold reduction of rivanol treatments, and a higher yield and purity of C-PC. Pure C-PC (A(620)/A(280) of 4.52) and APC (A(652)/A(280) of 2.41) were obtained. The estimated molecular masses of the alpha and beta subunits were 17 and 19 kDsmall a, Cyrillic for capital ES, Cyrillic-phycocyanin and 16 and 18 kDsmall a, Cyrillic for APC, respectively. The overall C-PC recovery of 55% (w/w) from its content (100 mg) in the crude extract was 10-20% higher than so far reported. The procedure appears promising for scaling up and broader applications.

Isolation and antibacterial activity of anabaena phycocyanin

“…It is consistent with that of spirulina (16 to 18 kDa) and arthronema africanum (17 to 19 kDa) (Liao and Zhang, 2011;Minkova et al, 2007). However, the PC-C was not certain in the electrophoresis graph.…”

Efficient separation and purification of allophycocyanin from Spirulina (Arthrospira) platensis

“…Gel filtration is a very useful method in phycobiliprotein separation and purification, but it is timeconsuming and not suitable for large-scale separation. A recently reported non-chromatographic, rivanol-sulfate method has been introduced and is fairly efficient with 35% recovery yield (Minkova et al 2007). Our hydroxylapatite extraction method is also easy and very efficient, with the recovery yield reaching as high as 70% in the separating step.…”

“…Previous studies on purifying phycobiliproteins have mainly focused on methods of purifying PE (Galland-Irmouli et al 2000;Rossano et al 2003;Bermejo et al 2003;Rito Palomares 2004, 2006;Liu et al 2005) or PC (Minkova et al 2003(Minkova et al , 2007Benedetti et al 2006;Patil et al 2006;Chen et al 2006;Zhu et al 2007;Silveira et al 2007), which are major phycobiliproteins in cyanobacteria or red algae, while publications on APC purification are much less (Bermejo et al 1997;Minkova et al 2007;. Conventional methodologies for purifying phycobiliproteins often involve a pretreatment step and several chromatography steps.…”

Large scale preparation of phycobiliproteins from <i>Porphyra yezoensis</i> using co-precipitation with ammonium sulfate

“…Besides ammonium sulfate, precipitation regents used in primary isolation include active carbon [28] and rivanol [29]. Therefore, we have used active carbon to absorb impurities in 8 http://www.iciba.com/grads/ gradients' solutions in which the active carbon to protein ratios (g/L) ranged from 1:1 to 50:1, resulting in the purity of 0.38 -0.53 and 0.16 -0.22, respectively, for PE and PC.…”

“…It was applied to isolate PBP from Porphyridium cruentum [30] and Nostoc sp [31]. CPC with purity of 3.90 and 3.00 from Spirulina (Arthrospira) fusiformis [32] and Arthronema africanum [29], respectively, were purified after fractional precipitation with rivanol. So we designed to use rivanol to absorb im-purities in 5 gradients' solutions in which rivanol to protein ratios (g/g) ranged from 1:10 to 16:10, and achieved a purity of 0.56 and 0.28 for PE and PC, respectively.…”