Variant Amylase Enzymes with Amino Acid Substitutions
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Solution Overview
Problem
Current industrial processes utilizing amylases face challenges with low activity, thermostability, acid tolerance, and pH stability, particularly under harsh conditions such as high temperatures and acidic environments, limiting their effectiveness in applications like starch processing, food, detergent, textile, paper, and biofuel industries.
Innovation Solution
Development of variant amylases with specific amino acid substitutions at positions such as 18, 19, 22, 35, 54, 71, 75, 130, 165, 175, 182, 187, 232, 233, 262, 271, 321, 325, 351, 372, 377, 378, 384, 392, 393, 394, 395, 417, and 420, which enhance their thermostability, acid tolerance, and pH stability, resulting in improved activity and stability compared to parent amylases.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If variant amylases with amino acid substitutions are developed to improve thermostability and acid tolerance, then stability under harsh conditions is enhanced, but enzyme complexity increases
Solution Approach 1:
The patent applies parameter changes by systematically modifying amino acid sequences at specific positions (18, 19, 22, 35, 54, 71, 75, 130, 165, 175, 182, 187, 232, 233, 262, 271, 321, 325, 351, 372, 377, 378, 384, 392, 393, 394, 395, 417, and 420) to enhance thermostability and acid tolerance. This involves changing the chemical parameters of the enzyme structure through targeted amino acid substitutions while maintaining overall enzyme function.
Solution Approach 2:
The patent implements local quality by introducing specific amino acid substitutions at predetermined positions within the enzyme sequence rather than uniform modifications throughout. This localized approach allows specific regions of the enzyme to acquire enhanced stability properties while preserving the overall catalytic function and minimizing structural complexity increases.
2Productivity
If amino acid substitutions are introduced to enhance activity and stability, then enzyme performance under harsh conditions improves, but manufacturing complexity increases
Solution Approach 1:
The patent changes the amino acid sequence parameters at specific positions to simultaneously improve amylase activity and stability. By targeting specific positions (such as 18, 19, 22, 35, 54, 71, 75, and others), the invention optimizes enzyme performance without requiring complete redesign of the manufacturing process.
Solution Approach 2:
The patent creates variant amylases that are copies of the parent enzyme with specific modifications. These variant enzymes replicate the core structure and function of the original while incorporating targeted amino acid substitutions that enhance performance, allowing for relatively straightforward production using existing manufacturing infrastructure.
3Stability of the object's composition
If multiple amino acid substitutions are made to improve pH stability and thermostability, then enzyme robustness increases, but the number of modifications required increases
Solution Approach 1:
The patent systematically changes amino acid parameters at multiple specific positions to achieve enhanced pH stability and thermostability. By selecting specific positions (18, 19, 22, 35, 54, 71, 75, 130, 165, 175, 182, 187, 232, 233, 262, 271, 321, 325, 351, 372, 377, 378, 384, 392, 393, 394, 395, 417, and 420), the invention achieves composition stability with a focused set of modifications rather than random extensive changes.
Solution Approach 2:
The patent segments the enzyme structure into specific modification positions and non-modification regions. By concentrating amino acid substitutions at predetermined positions while leaving other regions unchanged, the invention achieves enhanced stability without requiring comprehensive modification of the entire enzyme structure, thus reducing overall complexity.
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 variant amylases exhibit at least 1.1-fold better activity and stability under various conditions, including elevated temperatures and acidic pH, making them more suitable for diverse industrial applications.
Implementation Method 1
Amylases are enzymes that catalyze the hydrolysis of starch molecules to yield dextrins and oligosaccharides
Implementation Method 2
α-amylases (E.C.3.2.1.1) are enzymes that catalyses the hydrolysis of internal α-1,4-glycosidic linkages in starch
Data Source
AI summary
The invention is directed to novel amylases.