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Dextrin-uricase conjugate: Preparation, characterization, and enzymatic properties
Xiuting Hu1, Yu Wang2, Chengmei Liu1
1State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China; School of Food Science and Technology, Nanchang University, Nanchang 330047, China.
International Journal of Biological Macromolecules
|January 3, 2018
Summary
Conjugating uricase with dextrin enhances its stability and resistance to degradation. This modified uricase retains significant activity, even after being triggered by alpha-amylase, showing promise for therapeutic applications.
Area of Science:
- Biochemistry
- Enzyme Engineering
- Bioconjugation
Background:
- Uricase is an enzyme crucial for uric acid metabolism.
- Native uricase suffers from poor stability and rapid degradation in physiological conditions.
- Developing more stable uricase formulations is essential for therapeutic use.
Purpose of the Study:
- To enhance the stability of uricase through conjugation with dextrin.
- To investigate the impact of dextrin modification degree on uricase conjugation and activity.
- To evaluate the enzymatic properties and stability of the dextrin-uricase conjugate.
Main Methods:
- Succvinylation of dextrin to create dextrin monosuccinate with varying degrees of substitution (DS).
- Synthesis of dextrin-uricase conjugate using prepared dextrin monosuccinate and uricase.
- Characterization of the conjugate using size-exclusion chromatography and anion exchange chromatography.
- Assessment of enzyme activity recovery using alpha-amylase.
- Determination of optimal pH and temperature, and evaluation of stability against simulated physiological conditions and trypsin.
Main Results:
- Dextrin-uricase conjugate formation was confirmed, with conjugation degree increasing with dextrin monosuccinate DS.
- Conjugation led to initial activity loss, but significant recovery (83.4%) was achieved after alpha-amylase treatment at a 30:1 molar ratio (DS=0.283).
- The dextrin-uricase conjugate exhibited a slightly shifted optimal pH (9.0 vs 8.5) and enhanced stability under physiological conditions and against trypsin compared to free uricase.
Conclusions:
- Conjugation with dextrin monosuccinate effectively improves uricase stability.
- The degree of substitution in dextrin monosuccinate influences conjugation efficiency and activity recovery.
- Dextrin-uricase conjugate demonstrates superior resistance to degradation, offering a promising approach for enzyme stabilization.
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