Bacillus pumilus protease variants for cleaning stability
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Solution Overview
Problem
Existing proteases used in washing and cleaning agents often lack sufficient catalytic activity and storage stability, particularly in liquid surfactant-containing preparations, leading to suboptimal cleaning performance on protein-containing stains under standard washing conditions.
Innovation Solution
A protease from Bacillus pumilus with specific amino acid substitutions, such as 9T, 130D/V, 133A, 144K, 217M, 224A, 252T, and 271E, along with additional substitutions like 89A/G, 131H/Y/F, and 189T/L/I, is developed to enhance cleaning performance and stability, maintaining at least 70% sequence identity with the wild-type sequence.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional proteases are used in liquid surfactant-containing preparations, then the formulation is simple and easy to manufacture, but the proteolytic activity is insufficient and storage stability is poor under standard washing conditions
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid residues in the protease sequence (e.g., positions 9, 130, 133, 144, 217, 224, 252, 271) to alter the enzyme's physical and chemical properties. These sequence variations improve storage stability and proteolytic activity while maintaining the overall subtilisin structure, resolving the contradiction between stability and structural simplicity.
2Productivity
If conventional proteases are used in washing agents, then the formulation is straightforward, but the cleaning performance on protein-containing stains is suboptimal at standard washing temperatures
Solution Approach 1:
The patent modifies specific parameters of the protease molecule through amino acid substitutions at defined positions, enhancing catalytic activity and cleaning performance on protein stains. The sequence variations improve effectiveness at standard washing temperatures while maintaining manufacturability, addressing the contradiction between productivity and complexity.
3Productivity
If proteases are optimized for high catalytic activity in liquid formulations, then cleaning performance improves, but storage stability deteriorates
Solution Approach 1:
The patent carefully selects specific amino acid positions for modification to achieve a balance between catalytic activity and storage stability. By targeting particular residues (e.g., positions 9, 130, 133, 144, 217, 224, 252, 271) with specific substitutions, the invention optimizes both proteolytic activity and stability simultaneously, 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 proteolytic activity and increased storage stability, achieving enhanced cleaning performance on proteolytically sensitive stains across various temperature ranges and maintaining stability for extended periods in washing agents.
Implementation Method 1
They act as non-specific endopeptidases and hydrolyze any acid amide bonds that are inside peptides or proteins
Implementation Method 2
Proteases are some of the technically most important enzymes. They bring about the decomposition of protein-containing stains on the item to be cleaned
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, 224A, 252T and 271E at the positions corresponding to positions 9, 130, 133, 144, 217, 224, 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 89, 131 and 189, and to the production and use thereof. Such proteases exhibit very good cleaning performance.