Trypsin purification using magnetic particles of azocasein-iron composite

卷:226

Menezes Estevam Alves, Maria Helena ; Nascimento, Gabriela Ayres ; Cabrera, Mariana Paola ; da Cruz Silveri, Sara Isabel ; Nobre, Clarisse ; Teixeira, Jose Antonio ; de Carvalho Junior, Luiz Bezerra

Univ Fed Pernambuco

关键词: Enzyme purification;    Proteases;    Fish viscera;    Azocasein iron oxides;   

DOI  :  10.1016/j.foodchem.2016.12.094

学科分类：食品科学和技术

【 摘 要 】

This work presents an inexpensive, simple and fast procedure to purify trypsin based on affinity binding with ferromagnetic particles of azocasein composite (mAzo). Crude extract was obtained from intestines of fish Nile tilapia (Oreochromis niloticus) homogenized in buffer (01 g tissue/ml). This extract was exposed to 100 mg of mAzo and washed to remove unbound proteins by magnetic field. Trypsin was leached off under high ionic strength (3 M NaCl). Preparation was achieved containing specific activity about 60 times higher than that of the crude extract. SDS-PAGE showed that the purified protein had molecular weight (24 kDa) in concordance with the literature for the Nile tilapia trypsin. The mAzo composite can be reused and applied to purify trypsin from other sources. (C) 2016 Published by Elsevier Ltd.