Subtilisin Variants for Egg Yolk Stain Removal
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Solution Overview
Problem
Despite advancements in protease technology, there remains a need for improved proteases with enhanced properties such as wash performance, thermal stability, and catalytic activity to effectively remove tough stains like egg yolks from hard surfaces under varying washing conditions.
Innovation Solution
Development of subtilisin variants with specific amino acid substitutions, including positions 9{R,K,H}, 15{G,A,S,T,M}, 68{G,A,S,T,M}, 218{D,S,G,V}, and 245{R,K,H}, along with modifications like G61E, A98S, S99G, and N218D, to enhance wash performance, stability, and catalytic activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional proteases are used in detergent formulations, then basic washing function is provided, but wash performance on tough stains like egg yolks is insufficient
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid residues at positions 9, 15, 68, 218, and 245 of the subtilisin sequence to create variants with enhanced catalytic activity and stability. These parameter changes at the molecular level directly improve the enzyme's ability to remove tough stains like egg yolks while maintaining functional stability under varying washing conditions.
Solution Approach 2:
The invention applies local quality by introducing specific amino acid substitutions at targeted positions (9, 15, 68, 218, 245) rather than throughout the entire protein sequence. This localized modification approach optimizes specific functional regions of the subtilisin molecule to enhance stain removal capability while preserving the overall structural integrity and basic washing function.
2Reliability
If protease variants with enhanced catalytic activity are developed, then wash performance improves, but thermal stability may be compromised
Solution Approach 1:
The patent simultaneously optimizes multiple parameters by selecting specific amino acid substitutions that enhance both catalytic activity and thermal stability. The variants at positions 9, 15, 68, 218, and 245 are designed to improve enzyme activity while the structural modifications at these positions also contribute to maintaining thermal stability through enhanced protein folding and reduced conformational flexibility.
Solution Approach 2:
The invention creates composite enzyme variants by combining multiple amino acid substitutions at different positions within the subtilisin sequence. This composite approach allows the enzyme to integrate multiple functional improvements, where certain substitutions enhance catalytic activity while others contribute to thermal stability, achieving a balance between activity and stability.
3Reliability
If multiple amino acid substitutions are introduced to improve performance, then wash performance and stability improve, but protein engineering complexity increases
Solution Approach 1:
The patent reduces engineering complexity by focusing modifications on specific key positions (9, 15, 68, 218, 245) rather than throughout the entire sequence. This localized approach simplifies the protein engineering process by identifying and optimizing only the most critical residues that have the greatest impact on performance and stability.
Solution Approach 2:
The invention simplifies the engineering process by establishing a defined set of parameter changes at specific positions with predictable outcomes. By pre-identifying the key positions and their optimal amino acid substitutions, the patent reduces the complexity of screening and characterization that would be required if modifications were made throughout the entire protein sequence.
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 variants demonstrate improved stain removal capabilities, particularly for egg yolks, and increased stability, making them suitable for use in detergent compositions for enhanced cleaning performance.
Implementation Method 1
The present invention concerns variants of parent subtilisins... improved wash performance, such as improved stain removal capability... improved catalytic activity
Data Source
AI summary
The present invention relates to protease variants.The present invention also relates to polynucleotides encoding the variant protease variants and to nucleic acid constructs, vectors, and host cells comprising the polynucleotides, and methods of using the variant enzymes, such as, in laundry and detergent compositions.