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Enhancing Chymotrypsin Activity and Stability of Capillary Immobilized Enzyme Microreactors Using Zeolitic
Shuyi Zhang1, Yijia Gan1, Han Wang1
1Beijing Key Laboratory of Environmentally Harmful Chemical Analysis, College of Chemistry, Beijing University of Chemical Technology, Beijing 100029, P. R. China.
Analytical Chemistry
|May 23, 2024
Summary
This study introduces a new method for creating highly stable open-tubular immobilized enzyme microreactors (OT-IMERs) using DNA-enzyme composites and zeolitic imidazolate frameworks (ZIF-L). The novel OT-IMERs demonstrate enhanced enzyme activity, stability, and efficiency for applications like proteolytic digestion analysis.
Area of Science:
- Analytical Chemistry
- Materials Science
- Biochemistry
Background:
- Open-tubular immobilized enzyme microreactors (OT-IMERs) are crucial for rapid sample processing but face limitations in enzyme loading, sample interference, and stability.
- Traditional OT-IMER preparation methods often result in reduced enzyme activity and poor operational stability.
Purpose of the Study:
- To develop a novel strategy for preparing highly active and stable OT-IMERs by integrating DNA-enzyme composites with zeolitic imidazolate frameworks (ZIF-L).
- To enhance enzyme stability and performance in microreactor systems for advanced analytical applications.
Main Methods:
- Immobilization of single-stranded DNA-enzyme composites within capillaries.
- In situ encapsulation of composites using zeolitic imidazolate frameworks (ZIF-L) triggered by DNA phosphate groups and Zn²⁺.
- Characterization of enzyme activity, affinity, and stability (thermal, pH, organic solvent) of the prepared microreactors using chymotrypsin (ChT) and glucose oxidase (GOx).
- Application of the developed microreactors in proteolytic digestion analysis.
Main Results:
- The prepared Chymotrypsin@ZIF-L-IMER exhibited higher activity and affinity compared to free chymotrypsin and traditional ChT-IMER.
- Significantly improved thermal, pH, and organic solvent stability were observed for ChT@ZIF-L-IMER over multiple reaction cycles.
- Glucose oxidase@ZIF-L-IMER also demonstrated superior stability compared to free glucose oxidase and GOx-IMER.
- ChT@ZIF-L-IMER achieved efficient proteolytic digestion with a reduced reaction time.
Conclusions:
- The novel ZIF-L encapsulation strategy effectively enhances enzyme stability and activity in OT-IMERs.
- This method offers a versatile approach for synthesizing robust enzyme microreactors with broad applicability.
- The developed OT-IMERs show significant promise for capillary electrophoresis-based high-performance enzyme analysis and other biochemical assays.
