Proteomics Shows New Faces for the Old Penicillin Producer Penicillium chrysogenum
Optimization of the protein extraction protocol from mycelia of P. chrysogenum. (a) Extraction and purification of protein samples with ‘‘Clean-Up Kit” (GE Healthcare), based on Kniemeyer and coworkers  and silver stained. As sample buffer was used, 7 M urea, 2 M thiourea, 4% (w/v) CHAPS, 0.8% (v/v) IPG buffer pH 3–10 nonlinear (NL) (GE Healthcare), 40 mM Tris, 1 mM EDTA, and 20 mM DTT . (b) The extraction is based on the method described by Fernández-Acero and coworkers  by using mortar gridding and phosphate buffer, plus blue silver Coomassie colloidal staining . As sample buffer was used, 8 M urea, 2% (w/v) CHAPS, 0.5% (v/v) IPG buffer pH 3–10 NL (GE Healthcare), 20 mM DTT, and 0.002% bromophenol blue . Precision plus protein standards (Bio-Rad) were used as markers. The molecular mass is indicated in kilo-Daltons (kDa). Note the problematic regions observed with the extraction method A, which are highlighted by arrowed square brackets (left and bottom), Barreiro et al., 2011.
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