Stabilization of Industrial Enzymes Using Heterocyclic Compounds: Investigating the Effect of Indole and Trans-Chalcone on the Thermal Stability and Amorphous Aggregation of Bacillus amyloliquefaciens Alpha-Amylase
Subject Areas : biochemicalLeila Adibi 1 , Parichehreh Yaghmaei 2 , parvaneh Magami 3 , Azadeh Ebrahim-Habibi 4 *
1 - Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
2 - Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
3 - Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
4 - Biosensor Research Center, Institute of Cellular-Molecular Sciences of Endocrinology and Metabolism, Tehran University of Medical Sciences; Endocrinology and Metabolism Research Center, Institute of Clinical Sciences of Endocrinology and Metabolism, Tehran University of Medical Sciences, Tehran, Iran
Keywords: Industrial enzymes, heterocyclic compounds, indole, trans-chalcone, thermal stability, alpha-amylase, Bacillus amyloliquefaciens,
Abstract :
Objective: Proteins can undergo structural changes leading to amyloid or amorphous aggregates. Amorphous aggregates are particularly significant for industrial or therapeutic enzymes, as they may trigger adverse reactions in biological systems.
Alpha-amylase, a widely used industrial enzyme, was studied here to evaluate the stabilizing effects of indole and trans-chalcone on its thermal stability and amorphous aggregation.
Materials and Methods: The thermal stability of the enzyme was assessed by measuring its residual activity over 20 minutes at 60°C. Amorphous aggregation
was kinetically monitored via turbidity measurements using a spectrophotometer. Deamidation assays and Congo red tests were performed to evaluate covalent changes and amyloid formation, respectively. These experiments were conducted in the presence and absence of trans-chalcone and indole derivatives. The amorphous aggregation state was further analyzed using transmission electron microscopy (TEM).
Findings: Indole, a nitrogen-containing heterocyclic compound, demonstrated superior stabilization and anti-aggregation effects compared to trans-chalcone, a flavonoid precursor. Enzyme deamidation was significantly lower in the presence of indole. TEM images confirmed reduced amorphous particle aggregation in indole-treated samples.
Conclusion: Indole-based structures are proposed as promising candidates for designing compounds that inhibit amorphous aggregation in industrial enzymes.
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