Lipase Variants for Low Calcium Detergent Activity

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current lipases used in detergents have reduced activity at low calcium concentrations, leading to ineffective lipid stain removal and 'graying' of treated surfaces, as the catalytic site is shielded by a lid domain that requires activation at a water/lipid interface.

Innovation Solution

Development of lipase variants with specific mutations at positions 86, 87, 90, and 92, such as substitutions to D, E, Q, and C, which alter the activation mechanism and improve activity at low calcium concentrations, enhancing wash performance.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If lipase is used in detergent compositions, then lipid stain removal is improved, but at low calcium concentrations the lipase activity is reduced due to the lid domain shielding the catalytic site

Engineering Contradiction:

Improvelipid stain removal efficiencyVSAvoidlipase activity at low calcium concentrations

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent applies parameter changes by introducing specific amino acid substitutions at positions 86, 87, 90, and/or 92 in the lid domain sequence. These substitutions (such as I86D, E87C, I90Q, N92D) alter the physical-chemical properties of the lid domain, changing its conformational behavior and activation characteristics, thereby improving lipase activity at low calcium concentrations while maintaining effectiveness at higher concentrations

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent effectively extracts or removes the dependency on calcium for lid activation by introducing mutations that enable calcium-independent activation. The modified lid domain can open and activate the catalytic site without requiring calcium ions, thus taking out the harmful dependency on calcium concentration that limits lipase activity in low-calcium detergent formulations

Inventive Principle:

Principle #2Taking out (Extraction)

2Strength

If the lid domain shields the catalytic site, then the lipase is protected when inactive, but the catalytic activity is reduced at water/lipid interfaces without proper activation

Engineering Contradiction:

Improveprotection of catalytic siteVSAvoidcatalytic activity at water/lipid interface

Core Design Contradiction:

StrengthVSProductivity

Solution Approach 1:

The patent applies dynamics by introducing mutations that enhance the dynamic flexibility of the lid domain. The substitutions at positions 86, 87, 90, and/or 92 make the lid more prone to opening and adopting active conformations at water/lipid interfaces, allowing the enzyme to dynamically transition between protected and active states more effectively in response to interface conditions

Inventive Principle:

Principle #15Dynamics

Solution Approach 2:

The patent applies preliminary action by pre-configuring the lid domain through specific amino acid substitutions to be more readily activated. The mutations create a lid domain that is pre-positioned or pre-configured to open more easily upon contact with water/lipid interfaces, eliminating the need for extensive conformational changes or calcium-dependent activation steps

Inventive Principle:

Principle #10Preliminary action

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 lipase variants exhibit improved specific activity and wash performance compared to parent lipases, maintaining effectiveness even at low calcium levels, thereby enhancing detergent efficiency in lipid stain removal.

Implementation Method 1

the lid opens on contact with the interface

Methodology Applied

Scientific EffectConformational change:

Implementation Method 2

the catalytic site in many lipases is shielded by a lid domain

Methodology Applied

Scientific EffectProtein folding:

Implementation Method 3

Lipases are included in detergent compositions to increase wash performance and specifically to improve lipid stain removal

Methodology Applied

Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 4

Lipase activity

Methodology Applied

Scientific EffectHydrolysis: Hydrolysis

Data Source

PatentUS9909109B2Lipase variants and polynucleotides encoding same

Publication Date: 2018.03.06 NOVOZYMES AS

AI summary

The present invention relates to isolated lipase variants, comprising a substitution at one or more positions corresponding to positions I86D,E,N,Q, E87C, I90D,E,Q, and N92D,E,Q of the mature polypeptide of SEQ ID NO: 2, wherein the variant has lipase activity. In some embodiments the present invention relates to isolated polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; methods of producing the variants; and compositions comprising the variants. The present invention also relates to methods of obtaining lipase variants; methods of cleaning; and use of lipase variants for cleaning.