Fungal Serine Protease Variants for Detergent and Thermal Stability
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Solution Overview
Problem
There is a need for alternative serine proteases that are effective at low or moderate temperatures and stable in detergents with varying properties, as existing proteases face challenges in removing proteinaceous stains efficiently and maintaining stability during storage and use.
Innovation Solution
Development of serine protease variants from Fusarium equiseti RF6318 with specific amino acid substitutions, such as V208I, and additional changes at other positions, which enhance thermal stability and detergent stability while maintaining or improving wash performance compared to the wild-type protease.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing proteases are used for removing proteinaceous stains, then stain removal function is provided, but thermal stability and detergent stability are insufficient
Solution Approach 1:
The patent applies parameter changes by modifying amino acid residues at specific positions (e.g., position 208, 216, 223, 230, 236, 247, 252, 268, 281, 287, 288) in the protease sequence to alter the enzyme's thermal stability and detergent stability parameters while preserving its catalytic activity for stain removal
Solution Approach 2:
The patent applies local quality by making targeted amino acid substitutions at specific local positions within the protease molecule rather than global modifications, allowing localized structural changes that enhance stability without compromising overall enzymatic function
2Use of energy by moving object
If existing proteases are used at low temperatures, then energy consumption is reduced, but enzyme activity and stability deteriorate
Solution Approach 1:
The patent modifies the protease's thermal parameters through amino acid substitutions, enabling the enzyme to maintain stability across a broader temperature range including low temperatures, thus allowing energy-efficient cold wash applications without sacrificing enzyme reliability
3Reliability
If protease variants with multiple amino acid substitutions are created, then thermal stability and detergent stability are improved, but protein structure and function may be affected
Solution Approach 1:
The patent applies local quality by selecting specific amino acid positions for substitution that are strategically located to enhance stability without disrupting the core catalytic structure, maintaining local structural integrity while improving global stability properties
Solution Approach 2:
The patent carefully controls the parameters of amino acid substitutions to achieve desired stability improvements while staying within thresholds that preserve the protein's fundamental structure and catalytic function
Data Source
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AI summary
The present invention relates to fungal serine protease variants, which comprise an amino acid substitution of valine at position 208 of the parent Fusarium equiseti Fe_RF6318 serine protease, wherein the position of the substitution corresponds to the amino acid sequence of the mature Fe_RF6318 enzyme defined in SEQ ID NO: 2. The variants have improved thermal stability and/or detergent stability compared to the parent Fe_RF6318 enzyme. Preferably the substitution is V208I and more preferably the variants comprise additional amino acid changes which further increase the stability. Also disclosed are nucleic acid sequences encoding said protease variants as well as recombinant vectors and host cells for the production of the variants. The serine protease variants are applicable in laundry and dish-washing detergent compositions, in treating fibers, in treating wool, in treating hair, in treating leather, in treating food or feed, or in any application involving modification, degradation or removal of proteinaceous material.