Title
Quillaja bark saponin effects on Kluyveromyces lactis β-galactosidase activity and structure
Author
Cíntia Tiemi Misugi Kayukawa, Marco Aurélio Schüler Oliveira, Elaine Kaspchak, Heloisa Bruna Soligo Sanchuki, LucianaIgarashi-Mafra, Marcos R. Mafra
Year
2020
Journal
Food Chemistry
Abstract
Saponins are known for their bioactive and surfactant properties, showing applicability to the food, cosmetic and pharmaceutical industries. This work evaluated the saponins effects on Kluyveromyces lactis β-galactosidase activity and correlated these changes to the protein structure. Enzyme kinetic was evaluated by catalytic assay, protein structure was studied by circular dichroism and fluorescence, and isothermal titration calorimetry was used to evaluate the interactions forces. In vitro enzymatic activity assays indicated an increase in the protein activity due to the saponin–protein interaction. Circular dichroism shows that saponin changes the β-galactosidase secondary structure, favoring its protein-substrate interaction. Besides, changes in protein microenvironment due to the presence of saponin was observed by fluorescence spectroscopy. Isothermal titration calorimetry analyses suggested that saponins increased the affinity of β-galactosidase with the artificial substrate o-nitrophenyl-β-galactoside. The increase in the enzyme activity by saponins, demonstrated here, is important to new products development in food, cosmetic, and pharmaceutical industries.
Instrument
J-815
Keywords
Circular dichroism, Secondary structure, Tertiary structure, Chemical stability, Biochemistry, Food science