Abstract

VEGA-OLIVEROS, Carolina; CHEGWIN-ANGARITA, Carolina  and  ARDILA-BARRANTES, Harold D.. Conditions for protein analysis of Lentinuia edodes mycelium obtained from liquid state fermentation. Rev.Colomb.Quim. [online]. 2019, vol.48, n.3, pp.3-12. ISSN 0120-2804.  https://doi.org/10.15446/rev.colomb.quim.v48n3.74843.

Few are the investigations focused on the biotechnological production and the development of analytical tools about Lentinuia edodes, an edible mushroom which has a potential for being used in the development of nutraceutical products. For this reason, in this research, the production of biomass of the mushroom over time by liquid state fermentation (LSF) was studied. Then, the conditions that allow obtaining protein extracts for the application of tools for proteomic analysis were selected. Trichloroacetic acid (TCA) - acetone and TCA - acetone - phenol were the two protein extraction methods which were compared in terms of extraction yield and separation profiles in 1D (SDS-PAGE) and 2D (IEF-SDS PAGE) electrophoresis. The highest production of biomass and total protein content was obtained after 10 days of LSF. Protein extraction with TCA - acetone -phenol presented the highest yield, resolution and number of bands in 1D electrophoresis. In 2DE the two methods allowed the extraction of proteins with isoelectric points in pH 3-10 range but, the TCA - acetone - phenol method favored a differential extraction of proteins in the range of 33 to 113 kDa. These results constitute a first application of electrophoretic separation techniques for future proteomic studies.

Keywords : chloroacetic acid; phenol; electrophoresis; mushrooms; shiitake.

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