Modified Protease Variants for Enhanced Cleaning Performance
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Solution Overview
Problem
Existing proteases used in detergents and cleaning agents often lack sufficient catalytic activity and storage stability at standard washing temperatures (20°C to 40°C), resulting in suboptimal cleaning performance on protease-sensitive soils.
Innovation Solution
A protease from Bacillus pumilus with specific amino acid substitutions at positions 271, 9, and other positions, such as 18, 29, 48, 92, 101, 130, 131, 133, 144, 149, 156, 162, 166, 172, 192, 217, 224, and 252, enhancing proteolytic activity and storage stability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If wild-type proteases from Bacillus pumilus are used in detergents, then the formulation can be produced, but the proteases exhibit insufficient catalytic activity and storage stability at standard washing temperatures (20°C to 40°C)
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid substitutions at defined positions (271E, 9T/H/S/A, and additional positions from predefined lists) in the protease sequence. These sequence parameter modifications result in enhanced catalytic activity and storage stability at standard washing temperatures, directly resolving the contradiction between reliability and productivity for wild-type proteases.
2Productivity
If proteases are modified to improve catalytic activity, then cleaning performance increases, but the complexity of protein modification increases
Solution Approach 1:
The patent applies local quality by making targeted amino acid substitutions at specific positions (271, 9, and other predefined positions) rather than random or comprehensive modifications. This localized approach to protein engineering improves catalytic activity while maintaining manageable modification complexity, as only specific residues are changed according to predefined patterns.
3Productivity
If proteases are optimized for high catalytic activity, then cleaning performance improves, but storage stability deteriorates
Solution Approach 1:
The patent applies composite materials by creating protease variants that simultaneously incorporate multiple amino acid substitutions (e.g., 271E combined with 9T/H/S/A and additional positions) to achieve both high catalytic activity and storage stability. This composite approach integrates multiple modifying elements into a single protein structure, resolving the contradiction between productivity and reliability.
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 catalytic activity and storage stability, leading to enhanced cleaning performance on protease-sensitive soils, with activity levels exceeding 100% of wild-type proteases and maintaining stability for extended periods.
Implementation Method 1
They act as non-specific endopeptidases and hydrolyze any amide bonds located within peptides or proteins
Implementation Method 2
Proteases are among the most technically important enzymes. They break down protein-based 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) an amino acid substitution on the position corresponding to the position 271, and (b1) at least one other amino acid substitution on at least one of the positions corresponding to the positions 18, 61, 92, 99, 137, 149, 156, 159, 162, 166, 172, 192, 199, 217 or 265; and/or (b2) an amino acid substitution on the position corresponding to the position 9, and another amino acid substitution on at least one position corresponding to the positions 29, 48, 101, 130, 31, 133, 144, 217, 224 and 252. The invention also relates to the production and use thereof. Said type of proteases have a very good cleaning performance.