Novel metalloproteases

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Solution Overview

Problem

There is a need for novel proteases suitable for specific conditions and uses in industrial applications, as existing proteases do not adequately address requirements for stability and performance in detergent compositions.

Innovation Solution

Development of novel metalloprotease enzymes derived from Bacillus species, such as Anoxybacillus rupiensis, Lysinibacillus macroides, and Brevibacillus brevis, which exhibit proteolytic activity across a range of pH and temperature conditions, making them suitable for use in detergent compositions and various cleaning applications.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If existing proteases are used in detergent compositions, then cleaning function is provided, but stability under detergent conditions is insufficient

Engineering Contradiction:
ImprovestabilityVSAvoidperformance under specific conditions
Core Design Contradiction:
ReliabilityVSDifficulty of detecting and measuring

Solution Approach 1:

The patent applies parameter changes by modifying amino acid residues at specific positions (e.g., positions 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100) in the protease sequence to enhance stability under detergent conditions while maintaining cleaning activity across different pH and temperature ranges

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent creates composite enzyme formulations by combining modified protease variants with detergent components, creating a composite cleaning system that provides both stability and cleaning function. The modified proteases are designed to work synergistically with surfactants and other detergent ingredients

Inventive Principle:
Principle #40Composite materials

2Adaptability or versatility

If proteases are designed for broad pH and temperature activity, then versatility is improved, but optimization for specific conditions becomes difficult

Engineering Contradiction:
Improveactivity rangeVSAvoidcondition optimization
Core Design Contradiction:
Adaptability or versatilityVSManufacturing precision

Solution Approach 1:

The patent uses parameter changes by introducing specific amino acid substitutions at multiple positions in the protease sequence to create variants with optimized activity ranges. Each variant is tailored for specific pH and temperature conditions while maintaining overall versatility through the modular nature of the sequence modifications

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent applies dynamics by creating a family of protease variants with different stability and activity profiles, allowing selection of the optimal variant for specific application conditions. The dynamic approach enables adaptation to different detergent formulations and washing conditions

Inventive Principle:
Principle #15Dynamics

3Productivity

If novel metalloproteases are developed for detergent use, then cleaning performance is improved, but development complexity increases

Engineering Contradiction:
Improvecleaning efficacyVSAvoidenzyme development
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The patent systematically changes amino acid parameters at specific positions to improve cleaning performance. The structured approach to sequence modification, focusing on key regions of the protease, reduces development complexity by providing a rational design framework rather than random mutagenesis

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent uses copying by creating multiple variants based on a parent sequence, allowing systematic exploration of sequence-space to improve cleaning performance. The modular variant design enables efficient screening and selection of high-performance enzymes

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 novel metalloproteases demonstrate enhanced stability and cleaning performance in detergent compositions, effectively hydrolyzing proteins in the presence of surfactants and other components, thereby improving cleaning efficacy across different conditions.

Implementation Method 1

Metalloproteases (MPs) are among the hydrolases that mediate nucleophilic attack on peptide bonds using a water molecule coordinated in the active site

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Implementation Method 2

a divalent ion, such as zinc, activates the water molecule. This metal ion is held in place by amino acid ligands, usually 3 in number

Methodology Applied
Scientific EffectCatalysis: Catalysis

Data Source

PatentEP3260538B1Novel metalloproteases
Publication Date: 2021.04.14 DANISCO US INC
  • EP3260538B1 patent drawingFigure 1.1~1.2
  • EP3260538B1 patent drawingFigure 1.3~1.4
  • EP3260538B1 patent drawingFigure 1.5A~1.5B

AI summary

Aspects of the present compositions and methods relate to novel metalloproteases polynucleotides encoding the novel metalloprotease, compositions and methods for use thereof.