RP-II Protease Variants Low-Temperature Activity
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Solution Overview
Problem
Current proteases used in detergents and other applications lack improved properties such as increased activity at low temperatures, thermostability, and altered Ca2+ dependency, which are essential for enhanced performance in detergent formulations and other industrial processes.
Innovation Solution
Modification of the amino acid sequence of RP-II proteases based on their three-dimensional structure to create variants with altered properties, such as increased activity at low temperatures, thermostability, and altered Ca2+ dependency, by analyzing and modifying specific amino acid residues or structural regions identified through structural considerations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If wild-type RP-II proteases are used, then the enzyme maintains natural stability and activity, but it lacks improved properties such as increased activity at low temperatures, thermostability, and altered Ca2+ dependency
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid residues in the protease sequence to alter its physical-chemical properties. Site-directed mutagenesis is used to change parameters such as temperature stability, pH optima, and calcium dependency by substituting specific residues at defined positions in the protein sequence.
Solution Approach 2:
The patent implements local quality by making targeted modifications at specific locations in the protease molecule rather than global changes. Specific amino acid residues at defined positions are modified to confer local improvements in stability or activity while maintaining the overall protein structure and function.
2Reliability
If the amino acid sequence of RP-II proteases is modified to create variants with altered properties, then improved stability and activity are achieved, but the complexity of protein engineering and characterization increases
Solution Approach 1:
The patent applies segmentation by dividing the protease molecule into distinct functional regions and modifying specific segments independently. The protein sequence is divided into defined positions and regions, allowing targeted modification of specific segments to achieve desired properties while maintaining other functional regions unchanged.
Solution Approach 2:
The patent uses parameter changes to systematically modify protease properties by altering amino acid sequences at defined positions. This approach enables controlled changes to stability, activity, and other parameters through site-directed mutagenesis followed by systematic characterization.
Data Source
AI summary
The present invention relates to methods for producing variants of a parent RP-II protease and the variants having altered properties as compared to the parent RP-II protease.