Purificação de faseolamina de feijão branco (Pasheolus vulgaris L.) por cromatografia de troca iônica em coluna de criogel

Abstract

Phaseolamine is a protein of plant origin, found only in legumes such as white beans. It is a protein that has been widely used to reduce overweight and has also been the focus of many studies, as it has the ability to be an anti-hyperglycemic in patients with diabetes, justifying the need to obtain this protein with a high degree of purity. Supermacroporous cryogels can be used as chromatographic supports for protein purification, especially when it is desired to maintain the bioactivity of the target molecule. Thus, a polyacrylamide cryogel functionalized with polyethyleneimine (PEI) was developed to be used as an anion exchanger in phaseolamine adsorption studies under different conditions of pH (7,2, 7,8 and 8,4) and salt concentration [ TRIS-HCl] (20 mM, 60 mM and 100 mM). The cryogel functionalized with PEI (cryo-PEI) showed interconnected pores, uniform and spongy structure, whitish color. The cryo-PEI showed low flow resistance and permeability (Kw) of 5,03 × 10 -13 m 2 . The axial dispersion (DAX) results of the cryo-PEI were good, ranging from 2,75 × 10 -7 m 2 /s to 3,27 × 10 -6 m 2 /s. It was possible to observe the best conditions at pH 7,8 and salt concentration of 100mM TRIS-HCl for yield and purification factor, resulting in the highest purification factor (10,838) and highest yield (149,706%). This study demonstrated a successful purification method for capturing phaseolamine by ion exchange chromatography.