Question

How to extract microalga proteins without having chlorophyll or other pigments ?

Need to perform 2DE with Haematococcus pluvialis and Nannochloropsis salina

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  • Tomáš Hluska · Palacký University of Olomouc
    I'd suggest PVP or PVPP
    http://en.wikipedia.org/wiki/Polyvinylpyrrolidone
    http://en.wikipedia.org/wiki/Polyvinylpolypyrrolidone
    but I'm not sure whether are they used also for protein purification.
  • Dola Bhattacharjee · Government of India
    Dear Vincent
    Protein content of microalgal biomass is assessed either by
    (i) colorimetric methods (sensitive to interference), or
    (ii) by methods that measure the concentration of elemental nitrogen in biomass (sensitive to interference).
    The cells are to be conditioned (involving bursting of cells, physically or chemically) so that it fully releases intracellular proteins, prior to assessment. Hydrolytic enzymes and chemicals like SDS can be used. Milling a cell slurry with glass beads, exposure to ultrasonic frequencies etc. could be effective as well.

    Following references may be of some help to you
    1. Barbarino, E., Lourenço, S.O., 2005. An evaluation of methods for extraction and
    quantification of protein from marine macro- and microalgae. J. Appl. Phycol. 17, 447–460.
    2. Meijer, E.A., Wijffels, R.H., 1998. Development of a fast, reproducible and effective
    method for the extraction and quantification of proteins of micro-algae. Biotechnol. Tech. 12, 353–358.
    3. Chisti, Y., Moo-Young, M., 1986. Disruption of microbial cells for intracellular products. Enzyme Microb. Technol. 8, 194–204.

    Best
    Dola
  • Dola Bhattacharjee · Government of India
    Another one:
    http://cdn.intechopen.com/pdfs/28765/InTech-Protein_purification.pdf

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