Alpha-Amylase Variants With Lower Calcium Dependency and Stability

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Solution Overview

Problem

Current alpha-amylases used in industrial processes lack improved specific activity, tailored substrate specificity, enhanced thermal and pH stability, and reduced calcium dependency, which limits their effectiveness in applications such as starch conversion and cleaning.

Innovation Solution

Development of novel alpha-amylolytic variants with specific mutations in the amino acid sequence of parent alpha-amylases like SPEZYME Xtra or AmyS, altering properties such as substrate specificity, thermal stability, pH stability, and calcium dependency, to enhance their performance in starch processing and cleaning.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If current alpha-amylases are used in industrial processes, then basic starch conversion function is provided, but specific activity and substrate specificity are insufficient

Engineering Contradiction:
Improvespecific activityVSAvoidsubstrate specificity
Core Design Contradiction:
ProductivityVSReliability

Solution Approach 1:

The patent applies parameter changes by introducing specific amino acid substitutions at defined positions in the alpha-amylase sequence. These substitutions alter the enzyme's catalytic properties and substrate binding characteristics, thereby improving both specific activity and substrate specificity through controlled modification of molecular parameters

Inventive Principle:
Principle #35Parameter changes

2Reliability

If current alpha-amylases are used in industrial processes, then basic catalytic function is provided, but thermal stability and pH stability are insufficient

Engineering Contradiction:
ImprovestabilityVSAvoidthermal stability
Core Design Contradiction:
ReliabilityVSTemperature

Solution Approach 1:

The patent introduces amino acid substitutions at positions known to influence structural stability. These changes modify the enzyme's three-dimensional structure and chemical properties, enhancing resistance to thermal denaturation and pH extremes while preserving catalytic function

Inventive Principle:
Principle #35Parameter changes

3Productivity

If current alpha-amylases are used in industrial processes, then basic starch hydrolysis is achieved, but calcium dependency limits effectiveness

Engineering Contradiction:
ImproveeffectivenessVSAvoidcalcium dependency
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The patent modifies amino acid residues involved in calcium binding sites. These substitutions alter the enzyme's cofactor requirements, reducing dependency on calcium ions while maintaining or enhancing catalytic efficiency. This allows the enzyme to function effectively in calcium-limited environments

Inventive Principle:
Principle #35Parameter changes

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 mutated alpha-amylases exhibit improved specific activity, stability, and reduced calcium dependency, leading to enhanced performance in starch conversion, ethanol production, laundry washing, and dish washing, with increased efficiency and stability across various industrial applications.

Implementation Method 1

Alpha (α)-amylases (α-1,4-glucan-4-glucanohydrolases, E.C. 3.2.1.1) constitute a group of enzymes, which catalyze the hydrolysis of starch and other linear and branched 1,4-glucosidic oligo- and polysaccharides

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Data Source

PatentEP2447361B1Geobacillus stearothermophilus alpha-amylase (AMYS) variants with improved properties
Publication Date: 2014.10.08 DANISCO US INC
  • EP2447361B1 patent drawingFigure 1
  • EP2447361B1 patent drawingFigure 1
  • EP2447361B1 patent drawingFigure 1

AI summary

Described are variants of a parent alpha-amylase that exhibits an alteration in at least one of the following properties relative to said parent alpha-amylase: specific activity, substrate specificity, substrate binding, substrate cleavage, thermal stability, pH-dependent activity, pH-dependent stability, oxidative stability, calcium dependency, pI, and wash performance. The variants are suitable for starch conversion, ethanol production, laundry washing, dish washing, hard surface cleaning, textile desizing, and/or sweetener production.