|
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. |
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publisher |
AMER CHEMICAL SOC |
|
issn |
2168-0485 |
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year published |
2021 |
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volume |
9 |
|
issue |
24 |
|
beginning page |
8327 |
|
ending page |
8335 |
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digital object identifier (doi) |
10.1021/acssuschemeng.1c03173 |
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web of science category |
9 |
|
subject category |
Chemistry, Multidisciplinary; Green & Sustainable Science & Technology; Engineering, Chemical |
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unique article identifier |
WOS:000664614900032
|
