Alpha-amylase variants with enhanced solubility at pH 6.0
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Solution Overview
Problem
Commercially available alpha-amylases have limited solubility at low and high pH levels, such as pH 6.0 and 10.0, which restricts their usage in various industrial applications due to limited pH-range activity and solubility.
Innovation Solution
Introduction of amino acid modifications at specific positions in the alpha-amylase enzyme sequence, such as positions 23, 33, 181, 260, 272, 323, 349, 357, and 408, to enhance solubility at pH levels ≤6.0, including substitutions, insertions, and deletions, resulting in variant polypeptides with increased solubility compared to parent alpha-amylases.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If commercially available alpha-amylases are used, then enzyme activity is maintained, but solubility at low and high pH levels deteriorates
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid residues in the alpha-amylase enzyme sequence at positions 23, 33, 181, 260, 272, 323, 349, 357, and/or 408. These sequential modifications alter the enzyme's physical and chemical properties, specifically improving solubility at pH ≤6.0 while maintaining enzyme activity across a broad pH range from 4.0 to 12.0.
2Ease of manufacture
If amino acid modifications are introduced to improve solubility at pH ≤6.0, then solubility improves, but enzyme sequence complexity increases
Solution Approach 1:
The patent applies local quality by making targeted modifications at specific amino acid positions (23, 33, 181, 260, 272, 323, 349, 357, and/or 408) rather than altering the entire enzyme sequence. This localized approach improves solubility at pH ≤6.0 while preserving the overall enzyme structure and function, avoiding unnecessary complexity throughout the entire protein sequence.
Data Source
AI summary
The present invention relates to variants of an alpha-amylase which have an increased solubility at pH 6.0 compared to the parent alpha-amylase. The present invention also relates to methods of making the variant alpha-amylase and the use of the variant alpha-amylase in processing starch, cleaning or washing textiles, hard surfaces, or dishes, making ethanol, treating an oil well, processing pulp or paper, animal feed, syrup production, and preparing a dough or a baked product prepared from the dough.