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Immobilization of Cytochrome C by Benzoic Acid (BA)-Functional UiO-66-NO2 and the Enzyme Activity Assay

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Abstract

Recently, metal–organic frameworks (MOFs) are considered to be the moderate hosts for the bio-enzymes owing to their unique 3D pores and controllable surface affinity to the target molecules. In this work, the benzoic acid (BA)–modulated UiO-66-NO2 was introduced, and cytochrome c (Cyt C) was chosen as the target enzyme to evaluate the immobilization efficiency of the resulting UiO-66-NO2-BA. The immobilization conditions including pH, adsorption time, and temperature and the initial concentrations of BA were optimized. The adsorption kinetics and thermodynamics were analyzed to further explore the enhanced adsorption mechanism. It is worth noted that all the UiO-66-NO2-BA exhibited evidently enhanced adsorption capacities in comparison with the unmodified UiO-66-NO2 due to the formation of the chemical bonds between the UiO-66-NO2-BA and cytochrome C, indicating the positive roles of BA modification. Finally, the activities of the immobilized cytochrome C were assessed by using the catalytic oxidation of ABTS in the presence of H2O2, which reactions were also conducted over the free cytochrome C for comparison. The evidently improved stability under definite pH range, prolonged durability against the organic solvents, and the good reusability of the immobilized cytochrome C highlight the prospect applications of functional MOF immobilized enzymes in the practical catalytic reactions.

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Funding

The authors are very grateful for the financial support of LiaoNing Science and Technology Development Foundation Guided by Central government (2021JH6/10500141).

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  1. Peide An and Fenfen Zhu contributed equally to this wok.

Authors and Affiliations

  1. Department of Chemistry, College of Sciences, Northeastern University, Shenyang, 10819, People’s Republic of China

    Peide An, Fenfen Zhu, Shiji Liu, Xiaolin Zhou, Hao Meng & Xia Zhang

  2. Donggang No. 2 high school, Donggang, 118300, People’s Republic of China

    Chunfang Wang

  3. College of Pharmacy, Liaoning University, Shenyang, 110036, China

    Yufeng Liu

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  1. Peide An
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  2. Fenfen Zhu
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  3. Shiji Liu
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  4. Xiaolin Zhou
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Fenfen Zhu conceived and designed research. Peide An performed software analyses and the most writing work. All the authors have read and approved the manuscript.

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Correspondence to Xia Zhang.

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Highlights

1. It is the first time to use BA-functional UiO-66-NO2 in cytochrome C immobilization.

2. UiO-66-NO2-BA showed distinct enhanced immobilization efficiency for cytochrome C.

3. BA modification endows UiO-66-NO2 hierarchical pores and diminished particle size.

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An, P., Zhu, F., Liu, S. et al. Immobilization of Cytochrome C by Benzoic Acid (BA)-Functional UiO-66-NO2 and the Enzyme Activity Assay. Appl Biochem Biotechnol 194, 5167–5184 (2022). https://doi.org/10.1007/s12010-022-04018-z

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