Keratinase Mutant Thermal Stability via Site-Directed Mutation
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Solution Overview
Problem
The keratinase derived from Brevibacillus parabrevis has poor thermal stability, leading to enzyme deactivation at high temperatures, which limits its application in industrial processes such as washing and feed processing.
Innovation Solution
A keratinase mutant is created by mutating specific amino acids at positions 181, 217, and 236, resulting in improved thermal stability, with mutations such as asparagine to aspartate, tyrosine to serine, and serine to cysteine, enhancing the enzyme's half-life and optimal temperature.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If the keratinase is used at high temperatures for industrial applications, then the reaction efficiency and productivity are improved, but the enzyme rapidly deactivates due to poor thermal stability
Solution Approach 1:
The patent applies parameter changes by mutating specific amino acid residues (N181D, Y217S, S236C) in the keratinase protein sequence to alter its thermal stability parameters. These point mutations modify the enzyme's structural properties, enabling it to maintain activity at higher temperatures without rapid deactivation, thus resolving the contradiction between productivity and reliability.
2Duration of action of moving object
If the keratinase maintains high activity at elevated temperatures, then the half-life and operational duration are extended, but the enzyme structure becomes more prone to denaturation
Solution Approach 1:
The patent modifies the enzyme's structural parameters through site-directed mutagenesis at positions 181, 217, and 236. These parameter changes in the amino acid sequence enhance the enzyme's resistance to thermal denaturation, allowing it to maintain both extended half-life and structural integrity at elevated temperatures.
Data Source
AI summary
The invention relates to the technical field of industrial biotechnologies, and discloses a keratinase mutant with improved thermal stability and use thereof. The asparagine at position 181, the tyrosine at position 217, and the serine at position 236 in the keratinase derived from Brevibacillus parabrevis (CGMCC No. 10798) are engineered by site-direction mutation, and combined at random to obtain an enzyme with combined mutations. The invention realizes the remarkable improvement of the thermal stability of keratinase, and has good theoretical value and application prospect.