Biochemical Aspects of a Serine Protease from Caesalpinia echinata Lam. (Brazilwood) Seeds: A Potential Tool to Access the Mobilization of Seed Storage Proteins
Figure 1
Elution profile of CeSP purification. (a) Hydrophobic interaction chromatography, where a Hitrap Phenyl column was equilibrated with 50 mM phosphate buffer, pH 7.0, 1.0 M (NH4)2SO4, and proteins were eluted with (NH4)2SO4 (0.95, 0.25, and 0.13 M). (b) Anion exchange chromatography, where a Resource Q column was equilibrated with 50 mM Tris buffer, pH 7.5, and proteins were eluted with a linear NaCl (0 to 0.50 M) gradient. (c) Gel filtration, where a Superdex 75 column was equilibrated with 50 mM phosphate buffer, pH 7.0, 0.15 M NaCl, and proteins were eluted in the same buffer. Inset: molecular masses as function of protein elution. Standard proteins: BSA (67.0 kDa), ovalbumin (43.0 kDa), chymotrypsinogen A (25.0 kDa) and ribonuclease A (13.7 kDa). All fractions were monitored by the absorbance at 280 nm (●) and enzymatic activity (°).
