Engineering Cytochrome C with Quantum Dots and Ionic Liquids: A Win-Win Strategy for Protein Packaging against Multiple Stresses
authors Shet, SM; Thayallath, SK; Bisht, M; Pereira, MM; Coutinho, JAP; Kotrappanavar, NS; Mondal, D
nationality International
journal ACS SUSTAINABLE CHEMISTRY & ENGINEERING
author keywords Cytochrome C; Quantum dots; Ionic liquids; Multiple stresses; Protein engineering; Biocatalysis
keywords KINETIC ENHANCEMENT; ENZYMATIC-ACTIVITY; STABILITY; RESIDUES; ENZYMES
abstract Enhancing the structural stability and catalytic activity of Cytochorme c (Cyt C) against harsh process conditions would boost its use in biocatalysis. Herein, a new protein engineering strategy with improved efficacy is demonstrated through judicious task-specific functionalization of Cyt C with quantum dots (QDs) and ionic liquids (ILs). Mn2+ doped ZnS QD and ILs ([Cho][Ac]; [Cho][Dhp]) were concomitantly used to decorate Cyt C, which was characterized using various analytical tools. The peroxidase activity at room temperature of engineered Cyt C (Cyt C-QD-IL) increased markedly (1.2 to 3.5-fold) as compared to that for bare Cyt C, Cyt C with QD, and Cyt C with ILs. Further, Cyt C-QD-IL showed better catalytic activity under various stresses such as high temperature (110 degrees C), presence of a chemical denaturant (6 M GuHCl), high oxidative stress (30 min H2O2), and presence of proteases. Molecular docking results indicate that QD interacted with the active site of Cyt C and IL interacted with side chain amino acids via electrostatic and H-bonding interactions. Such favorable allosteric interactions might be behind the improved activity of Cyt C-QD-IL. The observed catalytic activity is in harmony with the structural stability of the protein as confirmed by UV-vis, ATR-IR, and CD analysis. Thus, the unveiled strategy represents an innovative dimension of protein packaging foreseeing the development of more robust biocatalysts that can be used at high temperatures.
publisher AMER CHEMICAL SOC
issn 2168-0485
year published 2021
volume 9
issue 24
beginning page 8327
ending page 8335
digital object identifier (doi) 10.1021/acssuschemeng.1c03173
web of science category 9
subject category Chemistry, Multidisciplinary; Green & Sustainable Science & Technology; Engineering, Chemical
unique article identifier WOS:000664614900032
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journal impact factor 7.632
5 year journal impact factor 7.741
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