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

Abstract 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.

Bibliographic Details

Main Authors:	Pesoti,Aline Regiele, Oliveira,Bruno Menezes de, Oliveira,Augusto Cesar de, Pompeu,Dávia Guimarães, Gonçalves,Daniel Bonoto, Marangoni,Sérgio, Silva,José Antonio da, Granjeiro,Paulo Afonso
Format:	Digital revista
Language:	English
Published:	Sociedade Brasileira de Ciência e Tecnologia de Alimentos 2015
Online Access:	http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612015000400588
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