Protease Sequence Variants for Detergent Storage Stability

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Solution Overview

Problem

Existing proteases in washing and cleaning agents, particularly textile washing agents, suffer from insufficient catalytic activity and storage stability, especially when exposed to protease-sensitive stains like blood, egg yolk, and milk, under standard washing conditions.

Innovation Solution

Introduction of specific amino acid substitutions, such as N121F, A194C, A209V, T218I, and N237P, into the protease sequence of Halalkalibacter okhensis Kh10-101 or similar proteases, enhancing their storage stability and proteolytic activity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If proteases are used in washing and cleaning agents, then cleaning performance on protease-sensitive stains is improved, but storage stability deteriorates

Engineering Contradiction:
Improvecleaning performanceVSAvoidstorage stability
Core Design Contradiction:
ReliabilityVSStability of the object's composition

Solution Approach 1:

The patent applies parameter changes by modifying the amino acid sequence of the protease through specific substitutions (N121F, A194C, A209V, T218I, N237P). These molecular-level parameter changes in the enzyme structure improve both storage stability and catalytic activity, resolving the contradiction between stability and performance.

Inventive Principle:
Principle #35Parameter changes

2Productivity

If proteases are stored at higher temperatures, then storage stability deteriorates, but catalytic activity improves

Engineering Contradiction:
Improvecatalytic activityVSAvoidstorage stability
Core Design Contradiction:
ProductivityVSStability of the object's composition

Solution Approach 1:

The patent modifies the protease's amino acid sequence parameters to create a more stable enzyme structure that maintains catalytic activity across a broader temperature range. The specific amino acid substitutions enable the enzyme to resist thermal denaturation while preserving its catalytic function.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent performs preliminary stabilization of the protease through amino acid modification before storage. By pre-modifying the enzyme structure with stabilizing substitutions, the protease is prepared to withstand storage conditions and maintain activity without requiring optimal temperature conditions throughout storage.

Inventive Principle:
Principle #10Preliminary action

3Reliability

If proteases are stored for prolonged periods, then storage stability deteriorates, but cleaning performance should be maintained

Engineering Contradiction:
Improvecleaning performanceVSAvoidstorage duration
Core Design Contradiction:
ReliabilityVSDuration of action of stationary object

Solution Approach 1:

The patent applies preliminary action by modifying the protease structure in advance through specific amino acid substitutions. This pre-modification creates a more stable enzyme that resists degradation during prolonged storage, ensuring cleaning performance is maintained over extended periods.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent provides beforehand cushioning against storage degradation by incorporating stabilizing amino acid substitutions that protect the protease from denaturation and loss of activity during long-term storage, thereby cushioning against the deterioration that would otherwise occur over time.

Inventive Principle:
Principle #11Beforehand cushioning (Prior cushioning)

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 modified proteases exhibit improved storage stability and proteolytic activity, maintaining higher residual activity and enhanced cleaning performance on protease-sensitive stains even after prolonged storage at temperatures ranging from 20°C to 40°C.

Implementation Method 1

They cause the degradation of protein-containing stains on the articles to be cleaned... hydrolyze any acid amide bonds within peptides or proteins

Methodology Applied
Scientific EffectProteolytic activity: Hydrolysis

Data Source

PatentUS20260028609A1Protease variants with improved storage stability
Publication Date: 2026.01.29 HENKEL KGAA

AI summary

The invention relates to proteases that exhibit proteolytic activity and comprise an amino acid sequence that is at least 70% identical to the amino acid sequence indicated in SEQ ID NO:1 over its entire length, wherein the proteases, in each case based on the numbering in accordance with SEQ ID NO:1, have at least one amino acid substitution selected from the group consisting of N121F, A194C, A209V, T218I, N237P and N237W in at least one of the positions corresponding to positions 121, 194, 209, 218 and 237. Such proteases are suitable for use in washing and cleaning agents, in particular textile washing agents, and have improved storage stability in comparison with a reference protease. The invention also relates to the use of said proteases and methods in which they are used, and to washing and cleaning agents containing said proteases, in particular textile washing agents.