A novel two-step purification process for highly stable C-phycocyanin of analytical grade purity and its properties

Efficient and economic purification of phycobiliproteins can be achieved by a novel relatively simple two-step process involving foam fractionation and ion exchange chromatography. Foam fractionation, which has not previously been used to concentrate phycobiliproteins, is a low-cost and environmentally friendly method that provides a significant volume reduction prior to the chromatography step. Two C-phycocyanin fractions with purities of 4.66 and 4.25 with slightly different characteristics and an allophycocyanin fraction with a purity of 3.23 were obtained. Both C-phycocyanins contain α-subunits of 15.0 kDa and β-subunits of 16.4 kDa, whereas the molecular weight of allophycocyanin is 15.5 kDa. The resulting C-phycocyanin retains its properties at pH in the range of 3–10, whereas strong alkaline pH leads to its rapid degradation. The purified protein is completely resistant to temperature changes in the range of 4 to 50 °C and loses only about 13% of its initial concentration during a 5 h incubation at 60 °C. Interestingly, purified C-phycocyanin is relatively resistant to photochemical degradation, as the loss in concentration after 10 h exposure to light is only about 14%. The most suitable storage conditions are temperature of 4 °C and pH in the range 4–5. The final product with an analytical purity greater than 4 is suitable for use in food, biomedicine and as a therapeutic agent.

• Foam fractionation and ion chromatography for the purification of phycobiliproteins.

• C-phycocyanin stable over a wide temperature and pH range without a stabilizing agent.

• C-phycocyanin of analytical purity for food, medical and pharmaceutical applications.