Protease Variants with Targeted Substitutions for Storage Stability
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Solution Overview
Problem
Existing proteases used in washing and cleaning agents lack sufficient catalytic activity and storage stability under standard washing conditions, leading to suboptimal cleaning performance on protease-sensitive stains.
Innovation Solution
A protease from Bacillus pumilus with specific amino acid substitutions, including 9T, 130D/V, 133A, 144K, 217M, 252T, and 271E, and optionally additional substitutions at positions 6W/F, 89A/G, 131H/Y/F, 166M/L/I, 189T/L/I, 211N/Q, and 224A/G, enhances storage stability and catalytic activity, making it suitable for use in washing and cleaning agents.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional proteases are used in washing and cleaning agents, then they provide basic proteolytic activity, but they lack sufficient storage stability and catalytic performance under standard washing conditions
Solution Approach 1:
The patent applies parameter changes by systematically modifying specific amino acid positions in the protease sequence. Seven key positions (9, 130, 133, 144, 217, 252, 271) were identified and modified with specific amino acid substitutions to enhance storage stability while maintaining catalytic activity under washing conditions
Solution Approach 2:
The patent implements local quality by making targeted amino acid substitutions at specific positions rather than random mutagenesis. Each position was carefully selected and modified with specific amino acids (e.g., position 9 with T, position 130 with D or V) to locally improve stability properties without altering the overall protein structure
2Productivity
If proteases are optimized for catalytic activity under washing conditions, then cleaning performance improves, but storage stability deteriorates
Solution Approach 1:
The patent simultaneously optimizes both catalytic activity and storage stability by modifying multiple amino acid parameters together. The coordinated changes at seven positions create a protease variant that maintains high catalytic performance during washing while exhibiting improved storage stability, resolving the trade-off between these two properties
3Adaptability or versatility
If proteases are used in liquid surfactant-containing preparations, then they can be applied in washing agents, but they exhibit insufficient catalytic performance and storage stability
Solution Approach 1:
The amino acid substitutions in the protease variant enhance its compatibility with liquid surfactant-containing preparations. The modified protease maintains structural integrity and activity in the presence of surfactants and other formulation components, enabling stable incorporation into washing agent formulations
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 increased catalytic activity, resulting in enhanced cleaning performance on protease-sensitive stains across various temperature ranges, particularly 20°C to 40°C.
Implementation Method 1
They act as non-specific endopeptidases and hydrolyze any acid amide bonds that are inside peptides or proteins
Data Source
AI summary
Proteases may include an amino acid sequence having at least 70% sequence identity with the amino acid sequence given in SEQ ID NO:1 over its entire length. The proteases may further have, in each case based on the numbering according to SEQ ID NO:1, amino acid substitutions 9T, 130D/V, 133A, 144K, 217M, 252T and 271E at the positions corresponding to positions 9, 130, 133, 144, 217, 252 and 271. The proteases may further include at least one further amino acid substitution at at least one of the positions corresponding to positions 6, 89, 131, 166, 189, 211 or 224, and to the production and use thereof. Such proteases exhibit very good stability with good cleaning performance.