Alpha-amylase variants for cold water starch removal

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Solution Overview

Problem

Current alpha-amylases used in detergents and cleaning processes have limited effectiveness at low temperatures, making it difficult to maintain stain removal performance and activity in cold water washing cycles.

Innovation Solution

Development of alpha-amylase variants with specific alterations at positions G304, W140, W189, D134, E260, F262, W284, W347, W439, W469, G476, and G477, which enhance activity and stability at low temperatures, improving wash performance and stability compared to parent enzymes.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Loss of energy

If alpha-amylase is used in low temperature washing, then energy consumption is reduced and environmental impact is lowered, but enzyme activity and wash performance deteriorate

Engineering Contradiction:
Improveenergy consumptionVSAvoidenzyme activity
Core Design Contradiction:
Loss of energyVSReliability

Solution Approach 1:

The patent applies parameter changes by modifying the amino acid sequence of alpha-amylase at specific positions (G304, W140, W189, D134, E260, F262, W284, W347, W439, W469, G476, and G477) to alter the enzyme's temperature-activity profile. These sequence modifications enable the enzyme to maintain high activity at low temperatures (5-35°C) while preserving stability, thus resolving the contradiction between energy efficiency and enzyme performance.

Inventive Principle:
Principle #35Parameter changes

2Reliability

If alpha-amylase temperature optimum is maintained for high activity, then wash performance is improved, but washing temperature must be increased which raises energy consumption

Engineering Contradiction:
Improvewash performanceVSAvoidenergy consumption
Core Design Contradiction:
ReliabilityVSLoss of energy

Solution Approach 1:

The patent changes the biochemical parameters of alpha-amylase through site-directed mutagenesis at 12 specific positions, fundamentally altering the enzyme's temperature-activity relationship. The modified enzyme achieves optimal activity at low temperatures (5-35°C) compared to conventional enzymes that require higher temperatures, thereby improving wash performance without increasing energy consumption.

Inventive Principle:
Principle #35Parameter changes

3Loss of energy

If conventional alpha-amylase is used in cold water washing, then energy savings are achieved, but stain removal effectiveness is insufficient

Engineering Contradiction:
Improveenergy savingsVSAvoidstain removal effectiveness
Core Design Contradiction:
Loss of energyVSProductivity

Solution Approach 1:

The patent modifies key parameters of alpha-amylase including amino acid composition at 12 specific positions, catalytic efficiency, and temperature optimum. These changes enable the enzyme to maintain high catalytic activity and stain removal effectiveness at cold water temperatures, resolving the contradiction between energy savings and cleaning performance.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent creates engineered copies of alpha-amylase with modified amino acid sequences that replicate and enhance the desired low-temperature activity profile. These engineered enzyme variants serve as improved copies that maintain the fundamental amylolytic function while optimizing performance for cold water washing applications.

Inventive Principle:
Principle #26Copying

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 variants demonstrate improved wash performance and stability at low temperatures, effectively removing starch-based stains in cold water washing, addressing the limitations of existing alpha-amylases.

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.

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Data Source

PatentUS20240247244A1Alpha-amylase variants
Publication Date: 2024.07.25 NOVOZYMES AS

AI summary

The present invention relates to variants of a parent alpha-amylase. 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.