One-step chromatographic procedure for purification of B-phycoerythrin from Porphyridium cruentum

Protein Expr Purif. 2016 Jul:123:70-4. doi: 10.1016/j.pep.2016.01.018. Epub 2016 Feb 3.

Abstract

B-phycoerythrin (B-PE) was separated and purified from microalga Porphyridium cruentum using one-step chromatographic method. Phycobiliproteins in P. cruentum was extracted by osmotic shock and initially purified by ultrafiltration. Further purification was carried out with a SOURCE 15Q exchange column and analytical grade B-PE was obtained with a purity ratio (A545/A280) of 5.1 and a yield of 68.5%. It showed a double absorption peaks at 545 nm and 565 nm and a shoulder peak at 498 nm, and displayed a fluorescence emission maximum at 580 nm. The analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed a bulky band between 18 and 20 kDa which could be assigned to subunits α and β and a low intensity band of 27 kDa assigned to γ subunit. Our protocol provides attractive alternative to consider for the purification procedure to obtain analytical grade B-PE at commercial level.

Keywords: B-phycoerythrin; Chromatography; Porphyridium cruentum; Purification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anion Exchange Resins / chemistry
  • Chromatography, Ion Exchange* / economics
  • Chromatography, Ion Exchange* / methods
  • Electrophoresis, Polyacrylamide Gel
  • Phycoerythrin / chemistry
  • Phycoerythrin / isolation & purification*
  • Porphyridium / chemistry*
  • Ultracentrifugation

Substances

  • Anion Exchange Resins
  • Phycoerythrin