Protease-Compatible Alpha-Amylase for Low-Temperature Cleaning
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Solution Overview
Problem
Existing cleaning agents struggle to effectively remove starch stains at low temperatures due to the instability of α-amylases in the presence of proteases, leading to reduced detergency and cleaning performance.
Innovation Solution
A mutant α-amylase with specific amino acid residue substitutions at positions E187, S241, N192, H240, and K278, enhancing stability and cleaning performance even at low temperatures when combined with protease in cleaning compositions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If conventional α-amylase is used in cleaning agents containing protease, then protease can be included for synergistic cleaning effect, but the stability of amylase is impaired
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of α-amylase through site-directed mutagenesis. Specific amino acid residues at positions 187, 241, 192, 240, and 278 are substituted to alter the enzyme's physical and chemical properties, thereby improving its stability in protease-containing cleaning agents while maintaining cleaning performance
Solution Approach 2:
The patent applies local quality by making targeted amino acid substitutions at specific positions (187, 241, 192, 240, and 278) in the α-amylase sequence. These localized changes at critical residues enhance the enzyme's resistance to proteolysis without compromising its overall catalytic function and starch-stain removal capability
2Loss of energy
If low temperature cleaning is used for environmental protection and cost reduction, then energy consumption is reduced, but amylase activity decreases because optimal temperatures are higher
Solution Approach 1:
The patent applies parameter changes by engineering the α-amylase to function optimally at lower temperatures. The amino acid substitutions modify the enzyme's thermal properties, allowing it to maintain high amylolytic activity at low cleaning temperatures (e.g., 20-40°C), thus enabling energy-efficient cleaning without sacrificing enzymatic performance
Solution Approach 2:
The patent applies dynamics by creating an α-amylase with adaptable temperature characteristics. The mutated enzyme can effectively operate across a broader temperature range, particularly excelling at low temperatures where conventional amylases would be inactive, providing flexible cleaning performance under varying environmental conditions
3Productivity
If combined use of α-amylase and protease is used for synergistic cleaning effect, then starch and protein stain removal is improved, but the stability of amylase in the cleaning agent is impaired
Solution Approach 1:
The patent applies parameter changes by modifying the α-amylase's amino acid sequence to increase its resistance to proteolytic degradation. The substitutions at positions 187, 241, 192, 240, and 278 create a more stable enzyme structure that can coexist with protease in the same cleaning formulation, enabling both enzymes to function synergistically without the amylase being degraded
Solution Approach 2:
The patent applies the intermediary principle by using the mutated α-amylase as a stable partner that can mediate synergistic action with protease. The engineered amylase serves as a reliable intermediary that maintains its structural integrity in the presence of protease, allowing both enzymes to work together on mixed starch-protein stains without one enzyme compromising the other
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 mutant α-amylase maintains high amylolytic activity and stability in protease-containing cleaning agents at low temperatures, ensuring effective starch stain removal and improved storage stability.
Implementation Method 1
α-amylases are known to have suitability for containing in cleaning agents, and are contained in dishwashing cleaning agents for automatic dishwashers and clothing cleaning agents as components removing starch stains
Implementation Method 2
the mutant α-amylase maintains high amylolytic activity and stability in protease-containing cleaning agents at low temperatures
Implementation Method 3
the combined use of protease causes the problem in that the stability of amylase in the cleaning agent is impaired
Implementation Method 4
the α-amylase mutant is a mutant of a parent α-amylase and comprises one or more amino acid residue substitutions at positions selected from the group consisting of positions corresponding to positions E187 and S241, and further one or more amino acid residue substitutions at positions selected from the group consisting of positions corresponding to positions N192, H240, and K278
Data Source
AI summary
Provided is an α-amylase-containing cleaning composition which has high amylolytic activity at low temperatures and also has high stability even in a protease-containing composition. A cleaning composition comprising an α-amylase mutant and protease, wherein the α-amylase mutant is a mutant of a parent α-amylase and comprises one or more amino acid residue substitutions at positions selected from the group consisting of positions corresponding to positions E187 and S241 in the amino acid sequence of SEQ ID NO: 2, and further one or more amino acid residue substitutions at positions selected from the group consisting of positions corresponding to positions N192, H240, and K278, wherein the parent α-amylase or α-amylase mutant has at least 85% sequence identity to the amino acid sequence of SEQ ID NO: 4.