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

Food Science and Technology (Campinas)

Aline Regiele Pesoti¹  Bruno Menezes De Oliveira¹  Augusto Cesar De Oliveira¹  Dávia Guimarães Pompeu¹  Daniel Bonoto Gonçalves¹  Sérgio Marangoni¹  José Antonio Da Silva¹  Paulo Afonso Granjeiro¹ 

关键词: purification;    characterization;    inhibitor of trypsin;    Chenopodium quinoa;    seeds;   

DOI  :  10.1590/1678-457X.6655

来源: SciELO

【 摘 要 】

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.

【 授权许可】

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