Alpha-Amylase Variants for Wash Stability and Catalytic Efficiency
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Solution Overview
Problem
Existing alpha-amylases used in industrial applications, such as detergents, lack improved properties such as wash performance, stability, and catalytic efficiency.
Innovation Solution
Development of alpha-amylase variants with specific deletions and alterations at defined positions, resulting in improved properties like increased wash performance, stability, and catalytic efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If alpha-amylases derived from microorganisms are used in industrial applications, then the enzymes can catalyze hydrolysis of starch and oligosaccharides, but the wash performance and stability are insufficient
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid residues at defined positions in the alpha-amylase sequence. Multiple variants are created with substitutions at positions such as 1, 2, 3, 4, 5, 9, 16, 17, 22, 23, 25, 28, 29, 30, 31, 32, 33, 35, 36, 37, 40, 42, 44, 48, 51, 54, 56, 60, 62, 70, 75, 82, 83, 84, 86, 87, 89, 90, 91, 93, 94, 95, 98, 105, 109, 113, 116, 118, 125, 128, 130, 131, 132, 134, 135, 136, 138, 140, 142, 144, 150, 151, 152, 154, 155, 158, 160, 165, 167, 169, 172, 174, 178, 186, 190, 192, 193, 195, 206, 208, 212, 214, 217, 218, 219, 222, 225, 227, 228, 229, 233, 235, 243, 246, 247, 250, 251, 252, 253, 255, 256, 260, 261, 263, 265, 267, 269, 275, 276, 280, 281, 282, 284, 285, 286, 288, 291, 296, 297, 299, 302, 303, 304, 306, 311, 312, 313, 315, 317, 319, 320, 321, 323, 324, 325, 328, 330, 334, 337, 338, 339, 343, 345, 346, 347, 355, 356, 360, 361, 371, 374, 375, 376, 377, 378, 379, 381, 382, 383, 384, 385, 387, 389, 391, 394, 395, 400, 402, 407, 408, 410, 411, 420, 423, 429, 430, 431, 437, 439, 442, 444, 445, 446, 449, 450, 451, 452, 454, 459, 460, 461, 463, 465, 467, 469, 470, 471, 473, 476, 477, 481, 482, and 485. These sequence modifications resolve the contradiction by improving both wash performance and catalytic efficiency simultaneously.
2Stability of the object's composition
If alpha-amylases are modified to improve wash performance, then the enzyme stability increases, but the sequence identity deviates from the parent enzyme
Solution Approach 1:
The patent applies local quality by making targeted amino acid substitutions at specific positions rather than throughout the entire sequence. The variants maintain high sequence identity (at least 60%, preferably at least 80%, more preferably at least 90%, and most preferably at least 95% identity) to the parent alpha-amylase while introducing localized modifications at defined positions to improve stability. This localized approach allows stability enhancement while preserving overall sequence similarity.
3Productivity
If multiple amino acid positions are altered to enhance catalytic efficiency, then the enzyme activity improves, but the structural complexity increases
Solution Approach 1:
The patent applies segmentation by dividing the alpha-amylase sequence into specific positions that can be independently modified. Rather than attempting to optimize the entire enzyme structure simultaneously, the invention focuses on altering amino acid residues at defined positions (such as positions 1, 2, 3, 4, 5, 9, 16, 17, 22, 23, 25, 28, 29, 30, 31, 32, 33, 35, 36, 37, 40, 42, 44, 48, 51, 54, 56, 60, 62, 70, 75, 82, 83, 84, 86, 87, 89, 90, 91, 93, 94, 95, 98, 105, 109, 113, 116, 118, 125, 128, 130, 131, 132, 134, 135, 136, 138, 140, 142, 144, 150, 151, 152, 154, 155, 158, 160, 165, 167, 169, 172, 174, 178, 186, 190, 192, 193, 195, 206, 208, 212, 214, 217, 218, 219, 222, 225, 227, 228, 229, 233, 235, 243, 246, 247, 250, 251, 252, 253, 255, 256, 260, 261, 263, 265, 267, 269, 275, 276, 280, 281, 282, 284, 285, 286, 288, 291, 296, 297, 299, 302, 303, 304, 306, 311, 312, 313, 315, 317, 319, 320, 321, 323, 324, 325, 328, 330, 334, 337, 338, 339, 343, 345, 346, 347, 355, 356, 360, 361, 371, 374, 375, 376, 377, 378, 379, 381, 382, 383, 384, 385, 387, 389, 391, 394, 395, 400, 402, 407, 408, 410, 411, 420, 423, 429, 430, 431, 437, 439, 442, 444, 445, 446, 449, 450, 451, 452, 454, 459, 460, 461, 463, 465, 467, 469, 470, 471, 473, 476, 477, 481, 482, and 485). This segmented approach to modification simplifies the complexity management while achieving improved catalytic efficiency.
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 alpha-amylase variants exhibit enhanced wash performance and stability, providing improved catalytic efficiency and substrate specificity compared to parent enzymes.
Implementation Method 1
Alpha-amylases (alpha-1,4-glucan-4-glucanohydrolases, E. C. 3.2.1.1) constitute a group of enzymes, which catalyses hydrolysis of starch and other linear and branched 1,4-gluosidic oligo- and polysaccharides.
Data Source
AI summary
The present invention relates to alpha-amylase variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.