Abstract
Stem bromelain, a widely accepted phytotherapeutical drug, exists as a partially folded intermediate at pH 2.0, condition also encountered in the gastrointestinal tract (its site of adsorption). Numerous studies have demonstrated that various environmental and intracellular factors affect the fibrillation property of stem bromelain, by accelerating the process of assembly. In this study, the effect of calf thymus DNA (CT-DNA) on stem bromelain is investigated using multiple approaches such as turbidity measurements, ANS binding assays, Rayleigh scattering, ThT binding, far-UV circular dichroism, dynamic light scattering fluorescence microscopy and transmission electron microscopy. CT-DNA induced large sized β-sheet aggregates of SB at pH 2.0, and propensity of aggregation concomitantly increases with increasing concentration of CT-DNA (0-100 μM) followed by saturation phase at higher CT-DNA concentration. Furthermore, isothermal titration calorimetric measurements suggested that electrostatic interaction between positively charged SB at pH 2.0 and negatively charged phosphate group of CT-DNA is the probable mechanism that leads to aggregate formation. However, the hydrophobic interaction between CT-DNA and SB cannot be neglected.
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