Extraction, purification and characterization of inhibitor of trypsin from Chenopodium quinoa seeds

Aline   Regiele Pesoti; Bruno   Menezes de Oliveira; Augusto Cesar   de Oliveira; Dávia   Guimarães Pompeu; Daniel   Bonoto Gonçalves; Sérgio   Marangoni; José Antonio   da Silva; Paulo Afonso  Granjeiro

Extraction, purification and characterization of inhibitor of trypsin from Chenopodium quinoa seeds

Autores

Aline Regiele Pesoti

Bruno Menezes de Oliveira

Augusto Cesar de Oliveira

Dávia Guimarães Pompeu

Daniel Bonoto Gonçalves

Sérgio Marangoni

José Antonio da Silva

Paulo Afonso Granjeiro

Editor

Sociedade Brasileira de Ciência e Tecnologia de Alimentos

Descripción

A novel trypsin inhibitor of protease (CqTI) was purified from Chenopodium quinoa seeds. The optimal extracting solvent was 0.1M NaCl pH 6.8 (p < 0.05). The extraction time of 5h and 90 °C was optimum for the recovery of the trypsin inhibitor from C. quinoa seeds. The purification occurred in gel-filtration and reverse phase chromatography. CqTI presented active against commercial bovine trypsin and chymotrypsin and had a specific activity of 5,033.00 (TIU/mg), which was purified to 333.5-fold. The extent of purification was determined by SDS-PAGE. CqTI had an apparent molecular weight of approximately 12KDa and two bands in reduced conditions as determined by Tricine-SDS-PAGE. MALDI-TOF showed two peaks in 4,246.5 and 7,908.18m/z. CqTI presented high levels of essential amino acids. N-terminal amino acid sequence of this protein did not show similarity to any known protease inhibitor. Its activity was stable over a pH range (2-12), temperatures range (20-100 °C) and reducing agents.