Enzyme Surface Charge Engineering for Detergent Stability

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Solution Overview

Problem

Current enzyme technologies face challenges in optimizing enzyme performance in detergent formulations due to suboptimal properties when enzymes are used outside their natural environment, leading to inefficient stain removal and stability issues in laundry detergents.

Innovation Solution

Engineering proteins, specifically enzymes like proteases and amylases, by altering their net surface charge and surface charge distribution to create variants with improved wash performance, stability, and solubility through targeted amino acid modifications and combinatorial mutagenesis.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If enzymes are used outside their natural environment in detergent formulations, then they can provide cleaning functionality, but their performance becomes suboptimal due to instability and improper charge characteristics

Engineering Contradiction:
Improveenzyme stabilityVSAvoidperformance in detergent
Core Design Contradiction:
ReliabilityVSEase of operation

Solution Approach 1:

The patent applies parameter changes by systematically modifying the net surface charge and charge distribution of enzymes through amino acid substitutions. This involves changing electrostatic parameters (surface charge density, charge polarity) to optimize enzyme performance in detergent environments, transforming the enzyme's interaction with detergent components and fabric surfaces.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent implements local quality by making targeted amino acid substitutions at specific surface locations of the enzyme. Rather than random mutagenesis, the method focuses on residues with solvent-accessible surfaces that directly interact with the detergent environment, locally modifying charge characteristics to improve stability and performance while preserving overall enzyme structure and function.

Inventive Principle:
Principle #3Local quality

2Productivity

If traditional protein engineering methods are used to optimize enzyme performance, then some improvement can be achieved, but the process is time-consuming and lacks systematic approach to electrostatic property optimization

Engineering Contradiction:
Improveenzyme performance optimization speedVSAvoidelectrostatic property control
Core Design Contradiction:
ProductivityVSManufacturing precision

Solution Approach 1:

The patent applies preliminary action by pre-identifying target amino acid residues based on their solvent accessibility and electrostatic properties before conducting mutagenesis. The method uses computational analysis to predict which surface residues will have the greatest impact on enzyme-detergent interactions, allowing focused engineering efforts on the most promising candidates and reducing the search space for optimization.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent implements feedback through iterative cycles of enzyme variant screening and performance evaluation. Each round of mutagenesis is followed by systematic testing of enzyme activity, stability, and charge characteristics, with results fed back into the design of subsequent mutation rounds. This feedback loop enables progressive optimization of electrostatic properties based on actual performance data.

Inventive Principle:
Principle #23Feedback

Data Source

PatentEP2171056B1Methods for improving protein performance
Publication Date: 2016.10.19 DANISCO US INC
  • EP2171056B1 patent drawingFigure 1A~1B
  • EP2171056B1 patent drawingFigure 2
  • EP2171056B1 patent drawingFigure 3A~3B

AI summary

The present invention provides methods for engineering proteins to optimize their performance under certain environmental conditions of interest. In some embodiments, the present invention provides methods for engineering enzymes to optimize their catalytic activity under particular environmental conditions. In some preferred embodiments, the present invention provides methods for altering the net surface charge and/or surface charge distribution of enzymes {e.g., metalloproteases or serine proteases) to obtain enzyme variants that demonstrate improved performance in detergent formulations as compared to the starting or parent enzyme.