Lipase Polypeptide Mutations for Detergent Wash Performance

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Solution Overview

Problem

Current lipase variants from Thermomyces lanuginosus lack optimal performance in removing long fatty acid chains while maintaining low odor generation and good wash performance in detergents.

Innovation Solution

Development of isolated polypeptides with specific mutations such as I202G + T231R + N233R, I86V + L227G + T231R + N233R + P256K, and others, which exhibit enhanced lipase activity and reduced odor potential, along with methods for their production and application in detergents.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional lipase variants from Thermomyces lanuginosus are used, then good wash performance is achieved, but long fatty acid chains are not effectively removed and odor generation occurs

Engineering Contradiction:
Improvewash performanceVSAvoidodor generation
Core Design Contradiction:
ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The patent applies parameter changes by introducing specific amino acid mutations at positions 202, 231, and 233 in the lipase protein sequence. These mutations alter the enzyme's substrate specificity and odor generation characteristics while maintaining wash performance. The mutations change the physical-chemical parameters of the enzyme active site to prefer long fatty acid chains and reduce odor-causing byproducts.

Inventive Principle:
Principle #35Parameter changes

2Object-generated harmful factors

If lipase variants with reduced odor potential are developed, then odor generation is decreased, but performance in removing long fatty acid chains may be compromised

Engineering Contradiction:
Improveodor potentialVSAvoidremoval of long fatty acid chains
Core Design Contradiction:
Object-generated harmful factorsVSProductivity

Solution Approach 1:

The patent applies local quality by making targeted mutations at specific positions (202, 231, 233) within the lipase protein structure. These localized changes in the active site region selectively modify substrate binding characteristics to favor long fatty acid chains while simultaneously reducing odor generation, rather than making global changes to the entire protein.

Inventive Principle:
Principle #3Local quality

3Reliability

If multiple mutations are introduced to improve lipase activity and reduce odor, then performance and odor control are improved, but protein stability and folding may be affected

Engineering Contradiction:
Improvelipase activityVSAvoidprotein stability
Core Design Contradiction:
ReliabilityVSStability of the object's composition

Solution Approach 1:

The patent applies segmentation by introducing mutations at three separate, spatially distributed positions (202, 231, 233) in the protein sequence. This segmented approach allows each mutation to contribute a specific function (substrate binding, odor reduction, or structural stabilization) while distributing the structural perturbation across multiple locations rather than concentrating it at one site, thereby maintaining overall protein folding and stability.

Inventive Principle:
Principle #1Segmentation

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

These variants demonstrate improved relative performance and reduced odor risk, achieving better wash performance and stability in detergent solutions.

Implementation Method 1

lipase activity is defined as a carboxylic ester hydrolase activity which catalyzes the hydrolysis of triacylglycerol under the formation of diacylglycerol and a carboxylate

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Implementation Method 2

The term 'lipase activity' is defined herein as a carboxylic ester hydrolase activity which catalyzes the hydrolysis of triacylglycerol

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Data Source

PatentEP1979476B1Polypeptides having lipase activity and polynucleotides encoding same
Publication Date: 2016.08.10 NOVOZYMES AS
  • EP1979476B1 patent drawingFigure 1
  • EP1979476B1 patent drawingFigure 1
  • EP1979476B1 patent drawingFigure 1

AI summary

The present invention relates to polypeptide having lipase activity and which further has a RP of at least 0.8 and a BR of at least 1.1 at the test conditions given in the specification.