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Quantification of Site-specific Protein Lysine Acetylation and Succinylation Stoichiometry Using Data-independent Acquisition Mass Spectrometry
Published on: April 4, 2018
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Stoichiometry and Kinetics of Sequential Dimethacrylate Enzymolysis.
S Frukhtbeyn1, K Van Dongen1, J Sun1
11 Volpe Research Center, American Dental Association Foundation, Frederick, MD, USA.
Journal of Dental Research
|July 23, 2019
Summary
This study developed methods to assess methacrylate degradation by enzymes. Cholesterol esterase proved more effective than pseudocholine esterase in breaking down dimethacrylates, highlighting the need for toxicity assessments of intermediate products.
Area of Science:
- Biomaterials Science
- Enzyme Kinetics
- Chemical Engineering
Background:
- Methacrylate-based materials are widely used in tissue engineering and dentistry.
- Concerns regarding toxicity, durability, and biocompatibility necessitate detailed evaluation of their degradation.
- Enzymolysis is a key degradation pathway requiring kinetic assessment.
Purpose of the Study:
- To develop tools for assessing and ranking the enzymolysis kinetics of dimethacrylate (DMA) compounds.
- To compare the efficacy of different esterases in DMA degradation.
- To investigate the hydrolysis resistance of intermediate mono-methacrylates (mono-MAs).
Main Methods:
- Kinetic studies of 2-step enzymolysis using pseudocholine esterase and cholesterol esterase on model DMAs (triethyleneglycol DMA, diurethane DMA).
- Quantification of enzymolysis products via high-performance liquid chromatography.
- Stoichiometric analysis and Berkeley Madonna modeling to compare esterase efficacy.
- Structural verification of mono-MAs using NMR spectroscopy and mass spectrometry.
Main Results:
- Cholesterol esterase demonstrated higher efficacy than pseudocholine esterase in sequential DMA degradation (>90% decomposition).
- Both enzymolysis steps (DMA to mono-MA, mono-MA to diol) followed first-order reaction kinetics.
- Mono-methacrylates exhibited greater resistance to hydrolysis compared to dimethacrylates.
- Stoichiometric analysis provided reliable conclusions on degradation pathways.
Conclusions:
- Stoichiometric analysis is a valuable tool for assessing DMA enzymolysis, especially when intermediates are difficult to quantify.
- The hydrolysis resistance of mono-MAs underscores the importance of their toxicity evaluation.
- Findings advocate for alternative material design strategies to enhance durability and biocompatibility.
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