Mannanase Variant Stability via Position 283 Substitution

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Solution Overview

Problem

Mannans in industrial applications, such as laundry and animal feed, pose challenges due to their high water binding capacity and viscosity, leading to issues like reduced digestibility and stability of mannanase enzymes, which affects performance in various processes.

Innovation Solution

Development of mannanase variants with a non-glycosylated amino acid at position 283, specifically designed to prevent N-linked glycosylation, enhancing stability and activity, and their use in enzyme compositions for improved performance in detergents and industrial processes.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If wild type mannanase is produced in a host cell capable of N-linked glycosylation, then the enzyme is produced with N-linked glycosylation at position 283, but this glycosylation reduces enzyme stability and specific activity

Engineering Contradiction:
Improveenzyme stabilityVSAvoidproduction yield
Core Design Contradiction:
ReliabilityVSEase of manufacture

Solution Approach 1:

The patent applies parameter changes by modifying the amino acid sequence at position 283 from asparagine (N) to threonine (T), thereby changing the glycosylation status of the enzyme. This substitution prevents N-linked glycosylation at the critical position 283, improving enzyme stability and specific activity while maintaining production feasibility in host cells

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent extracts the harmful glycosylation component by removing the N-linked glycosylation capability at position 283 through amino acid substitution. By eliminating this specific post-translational modification, the patent achieves improved enzyme stability and performance without affecting overall enzyme production

Inventive Principle:
Principle #2Taking out (Extraction)

2Productivity

If mannanase enzyme is used in laundry detergents, then it degrades mannan effectively, but the enzyme shows reduced stability in detergent conditions

Engineering Contradiction:
Improvemannan degradation activityVSAvoidenzyme stability in detergent
Core Design Contradiction:
ProductivityVSReliability

Solution Approach 1:

The patent changes the physical-chemical parameters of the enzyme by substituting amino acid residues at positions 283, 285, and/or 286, which alters the enzyme's interaction with detergent components and improves its stability under laundry conditions while preserving mannan degradation activity

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent applies local quality changes by making specific amino acid substitutions at positions 283, 285, and/or 286 in the enzyme sequence, thereby improving detergent stability at these specific locations without affecting the overall catalytic function of the enzyme

Inventive Principle:
Principle #3Local quality

3Quantity of substance

If mannan is present in animal feed, then it provides fiber intake, but it reduces feed digestibility and animal growth rate due to high viscosity

Engineering Contradiction:
Improvefiber content in feedVSAvoidfeed digestibility and growth rate
Core Design Contradiction:
Quantity of substanceVSProductivity

Solution Approach 1:

The patent uses mannanase enzyme as an intermediary substance that breaks down mannan in animal feed. By introducing this enzyme, the harmful high-viscosity mannan is degraded into smaller oligosaccharides, improving feed digestibility and animal growth while maintaining fiber intake benefits

Inventive Principle:
Principle #24Intermediary (Mediator)

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 variants exhibit improved stability and specific activity, leading to better performance in mannan degradation, increased yield, and enhanced applicability in laundry detergents and animal feed, improving digestibility and nutritional value.

Implementation Method 1

Mannanase increases average daily gain, feed efficiency, weight uniformity and livability in all monogastric animals

Methodology Applied
Scientific EffectEnzymatic hydrolysis: Hydrolysis

Data Source

PatentEP3409767B1Mannanase variants
Publication Date: 2020.10.21 AB ENZYMES OY
  • EP3409767B1 patent drawingFigure 1~2
  • EP3409767B1 patent drawingFigure 3A~3B
  • EP3409767B1 patent drawingFigure 4A~4B

AI summary

The present description is related to variants of mannanase, compositions comprising said variants, to methods for their production to methods of using said variants to degrade and modify mannan containing material.