Subtilase Variants for Detergent Storage Stability and Activity
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Solution Overview
Problem
Existing proteases used in detergent compositions face challenges such as enzyme instability due to varying washing conditions, leading to reduced wash performance and storage stability, necessitating improved proteases with enhanced stability and performance.
Innovation Solution
Development of subtilase variants with specific mutations at positions 209, 63, 215, and 217, including L209W, S63G or S63A, G215A, and Y217L, Y217I, Y217V, or Y217M, which maintain at least 80% sequence identity to SEQ ID NO: 1 and exhibit increased storage stability in detergent compositions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If wild-type subtilisin Carlsberg is used for detergent composition, then it provides baseline proteolytic activity, but it shows poor storage stability and forms aggregates during storage
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid residues at positions 31, 32, and 33 in the subtilisin Carlsberg sequence. These positional parameters were systematically varied to optimize both storage stability and proteolytic activity retention, resolving the contradiction between stability and activity.
Solution Approach 2:
The patent applies local quality by making targeted modifications at specific locations (positions 31-33) in the enzyme sequence rather than global changes. This localized approach allows optimization of storage stability in the N-terminal region while preserving the catalytic activity of the rest of the enzyme molecule.
2Productivity
If proteolytic enzymes are added to detergent composition, then they provide stain removal capability, but they lose activity rapidly during storage
Solution Approach 1:
The patent modifies amino acid parameters at positions 31-33 to create variants with enhanced durability. These parameter changes result in enzymes that maintain proteolytic activity for extended storage periods while retaining stain removal effectiveness in detergent applications.
Solution Approach 2:
The patent addresses the short-lived nature of wild-type enzymes by creating stabilized variants that function as durable, reusable components in detergent formulations, eliminating the need for frequent replenishment due to activity loss.
3Ease of manufacture
If wild-type subtilisin is used, then it is readily available through fermentation, but it forms aggregates and precipitates during storage
Solution Approach 1:
The patent applies local quality by modifying specific residues in the N-terminal region (positions 31-33) that are involved in aggregation-prone interactions. These localized changes improve solubility and prevent precipitation while maintaining the enzyme's overall structure and manufacturability through fermentation.
Solution Approach 2:
The patent systematically varies amino acid parameters at positions 31-33 to optimize solubility characteristics. These parameter changes reduce aggregation tendency during storage while preserving the enzyme's ability to be produced efficiently via fermentation processes.
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 mutated subtilase variants demonstrate improved storage stability, maintaining protease activity and enhancing wash performance under varying detergent conditions.
Implementation Method 1
The present invention relates to mutated subtilisin enzyme variants having improved stability and activity characteristics for use in aqueous detergent compositions
Data Source
AI summary
The invention relates to subtilase variants and detergent compositions comprising the variants, as well as methods of producing the variants and methods for stabilizing a subtilase variant.