Modified Protease Amino Acid Substitutions for Detergent Stability
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Solution Overview
Problem
Existing washing and cleaning agents with proteases often lack sufficient proteolytic activity under standard washing conditions, leading to suboptimal cleaning performance on protein-containing stains.
Innovation Solution
A protease from Bacillus pumilus with specific amino acid substitutions at positions P9, Q10, Q62, L82, P86, N130, T141, N187, S236, or T253, which enhances its catalytic activity and makes it more suitable for use in washing agents.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If standard proteases are used in washing and cleaning agents, then the formulations contain proteolytic activity, but the proteases do not have satisfactory levels of proteolytic activity under standard washing conditions
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the protease through site-directed mutagenesis. Specific amino acid residues at positions P9, Q10, Q62, L82, P86, N130, T141, N187, S236, and T253 are altered to optimize the enzyme's catalytic activity, stability, and substrate binding under washing conditions, thereby resolving the contradiction between reliability and productivity.
2Productivity
If proteases are altered by point, deletion or insertion mutagenesis to optimize performance, then catalytic activity may be improved, but the complexity of enzyme preparation and characterization increases
Solution Approach 1:
The patent applies local quality by implementing site-directed mutagenesis at specific positions (P9, Q10, Q62, L82, P86, N130, T141, N187, S236, T253) rather than random mutagenesis. This targeted approach modifies only the critical regions of the protease molecule that affect catalytic activity and stability, thereby improving productivity while minimizing the overall complexity of the optimization process.
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 increased proteolytic activity, providing improved cleaning performance on proteolytically sensitive stains across a wide temperature range, with activity levels up to 160% of the wild-type variant.
Implementation Method 1
They cause the breakdown of protein-containing stains on the item to be cleaned
Implementation Method 2
proteases are the longest established enzymes that are contained in virtually all modern, high-performance washing and cleaning agents
Data Source
AI summary
The present disclosure relates to proteases having an amino acid sequence with at least 70% sequence identity to the amino acid sequence given in SEQ ID No. 1, across its whole length, and comprising an amino acid substitution on at least one of the positions P9, Q10, Q62, L82, P86, N130, T141, N187, S236 or T253, relating in each case to the numbering according to SEQ ID No. 1. The present disclosure also relates to the production and use thereof. Said type of proteases have a very good cleaning performance.