Variant Alpha-Amylases for Extended Bread Freshness
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Solution Overview
Problem
Existing amylases do not effectively maintain bread freshness over an extended period, leading to staling due to starch crystallization, and their use in other industries is limited by inefficient exoamylase activity.
Innovation Solution
Development of variant alpha-amylases with increased exoamylase activity through targeted amino acid modifications, particularly at specific positions, enhancing starch degradation while maintaining product quality.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If existing amylases are used, then bread can be made, but the bread freshness is not maintained over an extended period due to staling
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the alpha-amylase enzyme to alter its catalytic properties. Specifically, mutations are introduced to increase exoamylase activity while maintaining or reducing endoamylase activity, thereby changing the enzyme's functional parameters to achieve prolonged bread freshness and reduced staling.
Solution Approach 2:
The patent applies local quality by making specific localized amino acid substitutions at particular positions in the enzyme sequence (e.g., positions 2, 3, 4, 21, 22, 25, 26, 29, 32, 35, 45, 53, 59, 68, 76, 82, 88, 90, 91, 96, 105, 117, 126, 128, 134, 141, 152, 160, 175, 197, 200, 234, 236, 243, 256, 257, 258, 261, 264, 270, 292, 311, 380, 416, 423, 433, 435). These localized modifications target specific functional regions to optimize exoamylase activity for anti-staling.
2Productivity
If existing amylases are used, then starch degradation occurs, but exoamylase activity is insufficient for efficient starch processing
Solution Approach 1:
The patent applies parameter changes by introducing amino acid substitutions that specifically enhance exoamylase activity. The mutations are designed to increase the enzyme's ability to perform exoamylase function (degradation of starch from chain ends) while maintaining overall alpha-amylase activity, thereby improving productivity in starch processing applications.
Solution Approach 2:
The patent applies local quality by targeting specific amino acid positions that are critical for exoamylase function. By making localized substitutions at these key positions, the enzyme's exoamylase activity is selectively enhanced, improving starch processing efficiency without requiring complete redesign of the enzyme structure.
3Reliability
If amino acid modifications are introduced, then exoamylase activity increases, but the enzyme sequence becomes more complex
Solution Approach 1:
The patent applies local quality by introducing a limited number of targeted amino acid substitutions at specific positions rather than making widespread modifications. This approach increases exoamylase activity while minimizing sequence complexity, as only essential residues are changed to achieve the desired functional improvement.
Solution Approach 2:
The patent applies parameter changes by making precise, minimal modifications to the amino acid sequence parameters. The substitutions are carefully selected to achieve the maximum increase in exoamylase activity with the minimum change in sequence complexity, maintaining ease of production and characterization.
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 alpha-amylases provide improved bread freshness and are applicable in various industries such as baking, detergents, personal care, and ethanol production, offering enhanced starch processing and viscosity control.
Implementation Method 1
amylase enzymes have been known to help maintain freshness over time (anti-staling or hardness) and maintain resilience overtime. The staling of bread is caused by the crystallization of amylopectin which takes place in starch granules after baking.
Implementation Method 2
The present inventors have surprisingly found that introducing amino acid modifications in the amino acid sequence of an alpha-amylase increases the exoamylase activity of the variant compared to the activity of the parent enzyme.
Data Source
AI summary
The present invention relates to variants of an alpha-amylase which have an increased exoamylase activity 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 baking, detergents, personal care products, in the processing of textiles, in pulp and paper processing, in the production of ethanol, lignocellulosic ethanol or syrups and as viscosity breaker in oilfield and mining industries.