Stability of the allergenic soybean Kunitz trypsin inhibitor

Authors

Robin Roychaudhuri, University of Nebraska-Lincoln
Gautam Sarath, University of Nebraska-LincolnFollow
Michael G. Zeece, University of Nebraska-LincolnFollow
John P. Markwell, University of Nebraska-LincolnFollow

Document Type

Article

Date of this Version

5-20-2004

Comments

Published in Biochimica et Biophysica Acta 1699 (2004) 207– 212.

Abstract

The soybean Kunitz trypsin inhibitor (SKTI) is a 21.5 kDa allergenic protein that belongs to the family of all antiparallel h-sheet proteins that are highly resistant to thermal and chemical denaturation. Spectroscopic and biochemical techniques such as circular dichroism (CD), ANS fluorescence and proteolysis were used to study its molecular structure under denaturing conditions such as acid and heat to which these allergens are commonly exposed during food processing. Reduction of native SKTI leads to its complete and rapid proteolysis by pepsin in simulated gastric fluid (SGF). Limited proteolysis with chymotrypsin during renaturation after heating showed that the native structure reforms at around 60 jC reversing the denaturation. CD spectra revealed that under acid denaturing conditions, SKTI shows major changes in conformation, indicating the possibility of a molten structure. The existence of this intermediate was established by ANS fluorescence studies at different concentrations of HCl. The remarkable stability of SKTI to both thermal and acid denaturation may be important for its role as a food allergen.