Protease Variants with Targeted Substitutions for Liquid Detergent Stability
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Solution Overview
Problem
Existing proteases used in detergents and cleaning agents often lack sufficient catalytic activity and storage stability, particularly in liquid surfactant-containing preparations, leading to suboptimal cleaning performance on protease-sensitive soils.
Innovation Solution
A protease with an amino acid sequence that has at least 80% sequence identity to a specific sequence, featuring targeted substitutions at positions 9, 271, and additional positions such as 29, 48, 101, 130, 131, 133, 144, 224, and 252, enhancing storage stability and catalytic activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If proteases are used in liquid surfactant-containing preparations, then cleaning performance on protease-sensitive soils can be achieved, but the proteases exhibit insufficient storage stability and catalytic activity
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the protease through site-directed mutagenesis. Specific amino acid substitutions at positions 9, 271, and other positions (29, 48, 101, 130, 131, 133, 144, 224, 252) are introduced to alter the enzyme's physical and chemical properties, thereby improving both storage stability and catalytic activity in liquid surfactant-containing preparations.
Solution Approach 2:
The patent applies local quality by making targeted local modifications to the protease structure through specific amino acid substitutions at defined positions. Rather than global changes, the invention focuses on localized sequence modifications at critical residues to optimize performance in liquid detergent formulations while maintaining overall enzyme functionality.
2Productivity
If standard proteases are used in detergents, then protein-containing soils can be degraded, but the proteases lack sufficient catalytic performance under washing conditions
Solution Approach 1:
The patent applies parameter changes by systematically modifying amino acid parameters at specific positions to enhance catalytic performance. The substitutions at positions 9 (e.g., P9T, P9H, P9S, P9A), 271 (Q271E), and other positions are designed to optimize the enzyme's catalytic efficiency, substrate binding, and stability under washing conditions while maintaining storage stability.
Solution Approach 2:
The patent applies self-service by engineering the protease to be self-stabilizing through intrinsic structural modifications. The amino acid substitutions create an enzyme variant that inherently maintains its catalytic activity and structural integrity in liquid surfactant environments without requiring external stabilizers or protective measures.
3Duration of action of stationary object
If proteases are stored in liquid formulations, then they can be used in cleaning agents, but they lose activity over extended storage periods
Solution Approach 1:
The patent applies preliminary action by pre-modifying the protease amino acid sequence before storage to prevent activity loss. The site-directed mutagenesis creates a stabilized enzyme variant that is pre-adapted to resist denaturation and maintain catalytic activity during extended storage in liquid formulations, eliminating the need for post-storage optimization.
Solution Approach 2:
The patent applies beforehand cushioning by introducing amino acid substitutions that act as protective measures against storage-induced degradation. The modifications at critical positions create structural resilience that cushions the enzyme against denaturation, aggregation, and activity loss during long-term storage in liquid detergent preparations.
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 protease exhibits improved storage stability and catalytic activity, maintaining effective cleaning performance on protease-sensitive soils even after extended storage periods and varying temperatures.
Implementation Method 1
They act as nonspecific endopeptidases and hydrolyze any acid amide bonds found within peptides or proteins
Implementation Method 2
Proteases are among the most technically important enzymes of all. They degrade protein-containing soils on the items being cleaned
Data Source
AI summary
The invention relates to proteases comprising an amino acid sequence which has at least 70% sequence identity to the amino acid sequence given in SEQ ID No.1 over its entire length, and (a) amino acid substitutions on the positions corresponding to the positions 9 and 271 relating in each case to the numbering according to SEQ ID No. 1, and (b) an amino acid substitution on at least one of the positions corresponding to the positions 29, 48, 101, 30, 131, 133, 144, 224 or 252, relating in each case to the numbering according to SEQ ID No.1. The invention also relates to the production and use thereof. Said type of proteases have a very good cleaning performance.