Strategy for a protein purification design using C-phycocyanin extract

Abstract

A variety of techniques have been developed for the separation and recovery of proteins. The cost of purifying the product is frequently determined by the desired quality of the final product, which is evaluated by measuring the purity. In this work the design of a protein purification process for C-phycocyanin, a phycobiliprotein that can be used in the food and medical industries, was established. The study evaluated the use of ammonium sulfate precipitation, ion exchange chromatography and gel filtration to purify C-phycocyanin in a variety of sequences. The final design included the C-phycocyanin extraction step, precipitation with ammonium sulfate and ion exchange chromatography. When the elution step was studied, the kind of elution and pH were considered in order to obtain a product with a final purity of 4.0 with a purification factor of 6.35, so that, at the end of the strategy, C-phycocyanin of analytical grade would be obtained.

Recovery of C-Phycocyanin in the Presence of Cells Using Expanded Bed IEC

“…Although the results obtained were in agreement with those found in the literature, Moraes and Kalil [18] [NaCl] Fig. 2 Chromatogram of C-phycocyanin purification in an expanded bed on Streamline QXL using gradient elution (0-1 M) in 150 mL, with a pH 6.5 buffer…”

“…The purity, concentration, recovery and purification factor were among the main responses that increased after optimizing the elution [18]. The variable pH is directly linked to purity, since the net negative charge of the protein varies according to the pH, binding more or less strongly onto the resin, which could lead to better adsorption of the target protein [18]. The elution volume promotes changes in the concentration and purity, since when the elution volume increases, there is a tendency for protein separation to improve, obtaining greater purity of the target molecule [19] but a reduced concentration of the eluate and consequently of the process recovery and yield.…”

“…The elution conditions were studied in this paper since this is a crucial step for more efficient purification. The purity, concentration, recovery and purification factor were among the main responses that increased after optimizing the elution [18]. The variable pH is directly linked to purity, since the net negative charge of the protein varies according to the pH, binding more or less strongly onto the resin, which could lead to better adsorption of the target protein [18].…”

Separation and purification of phycocyanin from Spirulina sp. using a membrane process

“…Many methods previously reported have been used to separate and purify phycocyanin from microalgae (Soni et al, 2006;Silveira et al, 2007;Singh et al, 2009). Precipitation with ammonium sulfate and the use of ultrafiltration (UF) can be used to obtain food grade phycocyanin, which has a purity ratio in the range of 0.7-2.0, whereas, gel filtration chromatography and ion-exchange chromatography can achieve analytical grade phycocyanin (Herrera et al, 1989;Rito-Palomares et al, 2001;Moraes and Kalil, 2009). Herrera et al (1989) reported that purification by UF, using a membrane with a molecular weight cut-off (MWCO) at 50 kDa, followed by activated charcoal adsorption and then spray drying, produced a food grade phycocyanin powder with a purity ratio of 0.74.…”

Application of prodigiosin as a colorant for polyolefines

“…The production of the coloration polymeric compositions by pigment preparation prodogiosin was described [9,10]. …”