Title
Folding and stability of recombinant azoreductase enzyme from Chromobacterium violaceum
Author
Kamalesh Verma, Debanjan Kundu, Lal Mohan Kundua, Ashish Kumar Singh, Vikash Kumar Dubey
Year
2019
Abstract
Azoreductase from Chromobacterium violaceum was characterized biophysically using experimental and computational tools. The in-silico docking and cross-linking experiments using glutaraldehyde suggest dimeric nature of the enzyme. The enzyme structure was modelled and also studied using circular dichroism (CD) spectroscopy which suggests 40% α- helix, 30% β- sheet and 30% random coils. In the modelled structure of the azoreductase, the cofactor flavin mononucleotide (FMN) binding energy was -3.8 kJ/mol. The binding of FMN affects the azoreductase-cofactor complex stability. The stability-folding studies indicate that the cofactor, FMN is required for folding, stability and activity. Overall, the data provides interesting insight into stability and biophysical parameters of the azoreductase protein.
Instrument
J-1500
Keywords
Circular dichroism, Secondary structure, Protein folding, Protein stability, Biochemistry