Conjugated microporous polymers-scaffolded enzyme cascade systems with enhanced catalytic activity

doi:10.1016/S1872-2067(24)60088-4

Abstract

Abstract:

Enhancing catalytic activity of multi-enzyme in vitro through substrate channeling effect is promising yet challenging. Herein, conjugated microporous polymers (CMPs)-scaffolded integrated enzyme cascade systems (I-ECSs) are constructed through co-entrapping glucose oxidase (GOx) and horseradish peroxidase (HRP), in which hydrogen peroxide (H₂O₂) is the intermediate product. The interplay of low-resistance mass transfer pathway and appropriate pore wall-H₂O₂ interactions facilitates the directed transfer of H₂O₂, resulting in 2.4-fold and 5.0-fold elevation in catalytic activity compared to free ECSs and separated ECSs, respectively. The substrate channeling effect could be regulated by altering the mass ratio of GOx to HRP. Besides, I-ECSs demonstrate excellent stabilities in harsh environments and multiple recycling.

Key words: Biocatalysis, Immobilization, Enzyme cascade system, Substrate channeling effect, Conjugated microporous polymers

Zhenhua Wu, Jiafu Shi, Boyu Zhang, Yushuai Jiao, Xiangxuan Meng, Ziyi Chu, Yu Chen, Yiran Cheng, Zhongyi Jiang. Conjugated microporous polymers-scaffolded enzyme cascade systems with enhanced catalytic activity[J]. Chinese Journal of Catalysis, 2024, 63: 213-223.

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URL: https://www.cjcatal.com/EN/10.1016/S1872-2067(24)60088-4

https://www.cjcatal.com/EN/Y2024/V63/I8/213

Figures/Tables 5

Fig. 1. Schematic illustration of the preparation of CMPs-scaffolded I-ECSs and the substrate channeling effect.

Fig. 2. Structural feature and characterization of I-ECSs. (a) SEM image of I-ECSs. TEM (b) and EDS elemental mapping images (c-f) of I-ECSs. CLSM images of FITC-labeled GOx (green) (g) and coumarin-labeled HRP (blue) (h). FTIR spectra (i) and pore size distribution (j) of CMPs and I-ECSs.

Fig. 3. Creation of substrate channeling effect in I-ECSs. Catalytic activity curves and τ of different biocatalyst concentrations biocatalysts: 2c (a), c (b), and 0.5c (c). (d) Reaction rate ratios of different concentrations biocatalysts. Catalytic activity curves (e) and reaction rate ratios (f) of I-ECSs and S-ECSs treatment with varying concentrations of free catalase (CAT). Reaction rate ratios calculated from vI-ECSs/vS-ECSs (vI-ECSs and vS-ECSs refer to the reaction rate of I-ECSs and S-ECSs, respectively). (g) The binding energy (ΔE) of pore wall-H2O2 via DFT, and the insert showed the hydrogen-bonded bridges between pore wall and H2O2. (h) Schematic illustration of I-ECSs (above) and the modelling of mass transfer pathway of H2O2 from producing sites to consuming sites (below). (i) Schematic illustration of S-ECSs (above) and the modelling of mass transfer pathway of H2O2 from producing sites to consuming sites (below).

Fig. 4. Fortification of substrate channeling effect in I-ECSs. (a) Catalytic activity curves and τ of I-ECSs-x with different dosages of HRP (I-ECSs-x represented the dosages of HRP in I-ECSs and τx represented τ of I-ECSs-x, x = 1, 3, and 5 mg). (b) Relative activity of I-ECSs-1, I-ECSs-3 and I-ECSs-5 treatment with 25 μg mL-1 free CAT. Reaction rate ratios of I-ECSs-3 or I-ECSs-5 and I-ECSs-1 treatment with varying concentrations of free CAT 0 (c) and 25 (d) μg mL-1, which was calculated from v3/v1 and v5/v1 (v1, v2, and v3 represented the reaction rate of I-ECSs-1, I-ECSs-3 and I-ECSs-5, respectively). (e) Catalytic activity curves of I-ECSs and free GOx-HRP cascade systems (F-ECSs). Notably, the catalytic activity of GOx or HRP in F-ECSs was equivalent to that of GOx or HRP in I-ECSs, obtained by regulating the mass of GOx or HRP in F-ECSs. (f) Reaction rate ratios of I-ECSs and F-ECSs.

Fig. 5. Tolerance of I-ECSs to inhospitable conditions. Stability of I-ECSs and F-ECSs to inhospitable conditions pH resistance (a), heating resistance (b), and proteolytic agent resistance (urea, acetone and N,N-dimethylformamide) (c). (d) Recyclability of I-ECSs.

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