Protease Mutations Enhance Low-Temperature Catalytic Activity
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Solution Overview
Problem
Proteases used in detergents and cleaning agents often lack sufficient catalytic activity and storage stability, particularly at low temperatures (20 °C to 40 °C), resulting in suboptimal cleaning performance on protein-containing dirt.
Innovation Solution
A protease from Bacillus pumilus with specific amino acid substitutions at positions N144 or S224, such as N144K, S224A, or S224T, which maintains at least 80% sequence identity to the wild-type sequence, exhibits enhanced catalytic activity and storage stability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If wild-type protease is used in detergents, then the formulation contains a protease with natural catalytic activity, but the protease lacks sufficient catalytic performance and storage stability at low temperatures (20°C to 40°C)
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid positions (N144, S224) in the protease sequence to alter its catalytic properties. The mutations N144K, S224A, and S224T were identified to enhance both catalytic activity and storage stability at low temperatures, directly resolving the contradiction between reliability and productivity.
Solution Approach 2:
The patent applies local quality by making targeted substitutions at specific positions (N144, S224) rather than global modifications. These localized changes in the amino acid sequence specifically improve low-temperature catalytic performance and storage stability while maintaining overall protease structure and function.
2Productivity
If proteases are optimized for catalytic activity, then washing performance improves, but storage stability decreases
Solution Approach 1:
The patent simultaneously optimizes both washing performance and storage stability through parameter changes at specific amino acid positions. The mutations N144K, S224A, and S224T were selected to improve both catalytic activity for washing performance and resistance to degradation during storage, eliminating the traditional trade-off between these properties.
Solution Approach 2:
The patent creates composite functional properties by combining multiple amino acid substitutions in the protease sequence. The combination of mutations at positions N144 and S224 produces a protease variant that exhibits both enhanced catalytic activity for washing and improved storage stability, achieving dual optimization.
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 demonstrates increased proteolytic activity of up to 160% compared to the wild-type variant, ensuring improved washing performance on proteolytic-sensitive soils and extended storage stability, even after several days.
Implementation Method 1
The modified protease demonstrates increased proteolytic activity of up to 160% compared to the wild-type variant, ensuring improved washing performance on proteolytic-sensitive soils
Data Source
AI summary
The invention 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, K22, V28, A29, A48, Q62, T78, L82, V84, S87, D101, N130, G131, T133, N144, G166, T215, S216, S224, N240, N252, S260 or Q271, 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.