Presenting B-DNA as macromolecular crowding agent to improve efficacy of cytochrome c under various stresses


Existence of numerous biomolecules results in biological fluids to be extremely crowded. Thus, Macromolecular crowding is an essential phenomenon to sustain active conformation of proteins in biological systems. Herein, double helical deoxyribonucleic acid (B-DNA) is presented for the first time as a biomacromolecular crowding system for sustainable packaging of cytochrome c (Cyt C). The peroxidase activity of Cyt C was investigated in the presence of various concentrations of B-DNA (from salmon milt). At an optimized concentration of 0.125 mg/ mL B-DNA, an 11-fold higher catalytic activity was found than in native Cyt C with improved stability. Molecular docking and spectroscopic analyses revealed that electrostatic and H-bonding are the main interactions between DNA and Cyt C that affect the structural stability and activity of the protein. Moreover, the catalytic activity and stability of the protein were further investigated in the presence of severe process conditions by UV-visible, circular dichroism, and Fourier-transform infrared spectroscopies. Molecularly crowded Cyt C showed significantly higher activity and stability under severe environments such as high temperature (110 degrees C), oxidative stress, high pH (pH 10) and biological (trypsin) and chemical denaturants (urea) compared to bare Cyt C. The observed results support the suitability of DNA-based macromolecular crowding media as a viable and effective stabilizer of proteins against multiple stresses.



subject category

Biochemistry & Molecular Biology; Chemistry; Polymer Science


Shet, SM; Bharadwaj, P; Bisht, M; Pereira, MM; Thayallath, SK; Lokesh, V; Franklin, G; Kotrappanavar, NS; Mondal, D

our authors


DM thanks SERB grant (EEQ/2021/000059) and RSC Research fund (R21-9087288636) for financial support. SKN expresses gratitude for Department of Science and Technology, Ministry of Science and Tech-nology, Government of India for Technology Mission Division grant (DST/TMD/HFC/2K18/124G) , DST-INSPIRE Faculty grant (IFA12-CH-84) and NANOMISSION grant SR/NM/NT-1073/2016.

Share this project:

Related Publications

We use cookies for marketing activities and to offer you a better experience. By clicking “Accept Cookies” you agree with our cookie policy. Read about how we use cookies by clicking "Privacy and Cookie Policy".