Engineering Cytochrome C with Quantum Dots and Ionic Liquids: A Win-Win Strategy for Protein Packaging against Multiple Stresses,ACS Sustainable Chemistry & Engineering

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Engineering Cytochrome C with Quantum Dots and Ionic Liquids: A Win-Win Strategy for Protein Packaging against Multiple Stresses
ACS Sustainable Chemistry & Engineering ( IF 7.1 ) Pub Date : 2021-06-04 , DOI: 10.1021/acssuschemeng.1c03173
Sachin M. Shet ₁ , Sarath Kumar Thayallath ₁ , Meena Bisht ₂ , Matheus M. Pereira ₂ , João A. P. Coutinho ₂ , Nataraj Sanna Kotrappanavar _{1,

3} , Dibyendu Mondal ₁

Affiliation

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 °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.

中文翻译：

用量子点和离子液体设计细胞色素 C：一种对抗多重压力的蛋白质包装的双赢策略

增强细胞色素 c (Cyt C) 对苛刻工艺条件的结构稳定性和催化活性将促进其在生物催化中的应用。在此，通过使用量子点 (QD) 和离子液体 (IL) 对 Cyt C 进行明智的任务特定功能化，证明了一种具有改进功效的新蛋白质工程策略。锰²⁺掺杂的 ZnS QD 和 IL（[Cho][Ac]；[Cho][Dhp]）同时用于装饰 Cyt C，并使用各种分析工具对其进行表征。与裸 Cyt C、带有 QD 的 Cyt C 和带有 ILs 的 Cyt C 相比，工程改造的 Cyt C (Cyt C-QD-IL) 在室温下的过氧化物酶活性显着增加（1.2 至 3.5 倍）。此外，Cyt C-QD-IL 在高温 (110 °C)、存在化学变性剂 (6 M GuHCl)、高氧化应激（30 分钟 H ₂ O ₂)，以及蛋白酶的存在。分子对接结果表明 QD 与 Cyt C 的活性位点相互作用，IL 通过静电和 H 键相互作用与侧链氨基酸相互作用。这种有利的变构相互作用可能是 Cyt C-QD-IL 活性提高的原因。观察到的催化活性与蛋白质的结构稳定性一致，这通过 UV-vis、ATR-IR 和 CD 分析得到证实。因此，公布的战略代表了蛋白质包装的创新维度，预见了可在高温下使用的更强大的生物催化剂的发展。

更新日期：2021-06-21

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