Variant Nitrile Hydratase Mutations for Stable Acrylamide Synthesis
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Solution Overview
Problem
Existing nitrile hydratase biocatalysts used for converting nitriles to amides face challenges with stability and activity, as they are deactivated by acrylonitrile and acrylamide, and modified biocatalysts often lack rapid conversion rates.
Innovation Solution
Engineering a novel nitrile hydratase with specific mutations in its alpha and beta subunits, optimized for enhanced stability and activity, and integrating it into suitable microorganisms for improved biocatalytic performance.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Temperature
If nitrile hydratase biocatalysts are used for converting nitriles to amides, then the conversion reaction can proceed under milder conditions compared to chemical synthesis, but the biocatalysts are deactivated by acrylonitrile and acrylamide, reducing stability
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid mutations (L6T, A19V, F126Y in alpha subunit; E108D, A200E in beta subunit) to modify the biocatalyst's physical and chemical properties. These mutations change the enzyme's structure to enhance its resistance to deactivation by acrylonitrile and acrylamide, thereby improving stability while maintaining the mild reaction conditions
2Reliability
If nitrile hydratase biocatalysts are modified to enhance stability, then resistance to deactivation by acrylonitrile and acrylamide is improved, but the activity and rapid conversion rate are reduced
Solution Approach 1:
The patent uses parameter changes by introducing specific amino acid mutations that simultaneously improve both stability and activity. The mutations L6T, A19V, F126Y in the alpha subunit and E108D, A200E in the beta subunit are designed to enhance resistance to deactivation while maintaining or improving catalytic activity, resolving the trade-off between stability and productivity
3Productivity
If unmodified nitrile hydratase biocatalysts are used, then high activity and rapid conversion of acrylonitrile to acrylamide is achieved, but the biocatalysts lack stability and are quickly deactivated
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid mutations (L6T, A19V, F126Y in alpha subunit; E108D, A200E in beta subunit) that modify the enzyme's structure to enhance its resistance to deactivation. These changes allow the biocatalyst to maintain high conversion rates while significantly improving stability compared to unmodified biocatalysts
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 modified nitrile hydratase exhibits increased stability and rapid conversion of acrylonitrile to acrylamide, suitable for industrial production with high yield and stability under reaction conditions.
Implementation Method 1
nitrile hydratase, a microbial enzyme that hydrolyses nitriles to amides
Implementation Method 2
nitrile hydratase producing microorganisms as biocatalysts
Data Source
AI summary
The present invention relates to a variant nitrile hydratase which is engineered to comprise greater activity and/or stability, nucleic acids encoding said nitrile hydratase, and microbia engineered to express said novel nitrile hydratase. Additionally the invention relates to the use of this nitrile hydratase and microbia which express said nitrile hydratase as a biocatalyst, particularly in methods for producing an amide compound from a nitrile compound, preferably for use in converting acrylonitrile to acrylamide.