Thiol-Based Deep Eutectic Solvent for Sortase Stability
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Solution Overview
Problem
Current deep eutectic solvents are limited in their ability to facilitate sortase-mediated bioconjugation reactions due to hydrolysis side reactions and instability of sortases in aqueous and organic solvents, which hampers the efficient conjugation of hydrophobic substrates and stability of enzyme intermediates.
Innovation Solution
Development of thiol-based deep eutectic solvents comprising a hydrogen bond acceptor and a hydrogen bond donor with thiol groups, which suppress hydrolysis side reactions and stabilize sortase enzymes, enabling efficient transpeptidation reactions in a water-free environment.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of substance
If water-free deep eutectic solvents are used, then hydrolysis side reactions are suppressed, but sortase enzyme stability decreases
Solution Approach 1:
The invention changes the chemical composition parameters of the deep eutectic solvent by incorporating specific thiols (such as ethanethiol, propanethiol) and hydroxyl compounds (such as ethanol, propanol) in optimized ratios. This parameter optimization allows the solvent to simultaneously suppress hydrolysis while maintaining sortase enzyme stability through favorable solvation effects.
Solution Approach 2:
The invention creates a composite deep eutectic solvent system combining multiple components: hydrogen bond acceptors (choline chloride, ethylene amine salts), hydrogen bond donors with thiol groups (ethanethiol, propanethiol), and hydroxyl compounds (ethanol, propanol). This composite formulation achieves synergistic effects where each component contributes to both hydrolysis suppression and enzyme stabilization.
2Loss of substance
If conventional deep eutectic solvents are used, then hydrolysis is suppressed, but solubility of hydrophobic substrates is insufficient
Solution Approach 1:
The invention optimizes the hydrophobicity parameters of the deep eutectic solvent by selecting specific thiol compounds with varying carbon chain lengths (ethanethiol, propanethiol) and adjusting their ratios. This parameter tuning enhances the solvent's ability to dissolve hydrophobic substrates while maintaining hydrolysis suppression through the water-free formulation.
Solution Approach 2:
The composite solvent system combines hydrophobic thiols with hydrophilic hydroxyl compounds and ionic components, creating a amphiphilic environment that can simultaneously accommodate hydrophobic substrates and maintain low water activity to suppress hydrolysis.
3Loss of substance
If organic solvents are used, then hydrolysis is suppressed, but sortase enzyme activity decreases
Solution Approach 1:
The invention changes the solvent class from conventional organic solvents to a specifically formulated deep eutectic solvent with optimized thiol and hydroxyl content. This parameter change creates a reaction environment that is sufficiently anhydrous to suppress hydrolysis but has appropriate polarity and hydrogen bonding capacity to maintain sortase enzyme activity.
Solution Approach 2:
The composite deep eutectic solvent combines ionic components (for structure and stability), thiol compounds (for hydrolysis suppression and enzyme stabilization), and hydroxyl compounds (for solubility and enzyme compatibility), creating a synergistic system that overcomes the limitations of simple organic solvents.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The thiol-based deep eutectic solvents enhance the stability of sortase enzymes and reduce hydrolysis, allowing for effective conjugation of hydrophobic substrates and improved reaction yields in bioconjugation processes.
Implementation Method 1
DESs are based on hydrogen bonds
Implementation Method 2
deep eutectics solvents can be produced water free and are therefore inert for hydrolysis
Implementation Method 3
enabling efficient transpeptidation reactions
Implementation Method 4
transpeptidation reaction can be performed in a thiol-based deep eutectic solvent
Data Source
AI summary
Herein is reported a deep eutectic solvent consisting of (2-hydroxyethyl) trimethyl ammonium chloride and dithiothreitol in a molar ratio of from 1:2 to 1:3 and of from 0 % to 10 % co-solvent.