Buttiauxella Phytase Variants for Thermal Stability

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Solution Overview

Problem

Monogastric animals such as pigs, poultry, and fish cannot metabolize phytate, leading to phosphorous pollution and the need for costly inorganic phosphate supplements, which also pose nutritional challenges due to phytic acid's antinutritional effects.

Innovation Solution

Development of a phytase variant with improved thermal activity and stability, specifically designed for animal feeds, which hydrolyzes phytate to release inorganic phosphate, enhancing phosphate digestion and reducing environmental pollution.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If inorganic phosphate is added to animal diets to provide sufficient phosphates, then phosphate availability for growth and health is improved, but cost increases and phosphorous pollution worsens

Engineering Contradiction:
Improvephosphate availabilityVSAvoidphosphorous pollution
Core Design Contradiction:
Quantity of substanceVSObject-generated harmful factors

Solution Approach 1:

The patent applies parameter changes by modifying the thermal stability parameter of phytase through site-directed mutagenesis. Specific amino acid substitutions (e.g., S75P, Q76R, A374P) are introduced to enhance the enzyme's resistance to thermal denaturation, allowing it to maintain activity at higher temperatures during feed processing and storage, thereby improving phosphate release efficiency without requiring additional inorganic phosphate

Inventive Principle:
Principle #35Parameter changes

2Quantity of substance

If phytase is used to hydrolyze phytate, then organic phosphorus availability is improved and phosphate pollution decreases, but enzyme stability under processing conditions deteriorates

Engineering Contradiction:
Improveorganic phosphorus availabilityVSAvoidenzyme stability
Core Design Contradiction:
Quantity of substanceVSReliability

Solution Approach 1:

The patent modifies the thermal stability parameter of phytase through site-directed mutagenesis. Specific amino acid substitutions (e.g., S75P, Q76R, A374P) are introduced to enhance the enzyme's resistance to thermal denaturation, allowing it to maintain activity at higher temperatures during feed processing and storage

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent applies local quality by making specific localized changes to the phytase protein structure at critical positions. Site-directed mutagenesis targets specific amino acid residues (such as positions 75, 76, and 374) to introduce substitutions that locally enhance thermal stability without altering the overall enzyme function or structure

Inventive Principle:
Principle #3Local quality

3Productivity

If phytase variant with improved thermal activity is developed, then phosphate digestion is enhanced, but manufacturing complexity increases

Engineering Contradiction:
Improvephosphate digestion efficiencyVSAvoidmanufacturing complexity
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The patent modifies the thermal stability parameter of phytase through site-directed mutagenesis. Specific amino acid substitutions (e.g., S75P, Q76R, A374P) are introduced to enhance the enzyme's resistance to thermal denaturation, allowing it to maintain activity at higher temperatures during feed processing and storage

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent applies universality by creating phytase variants that maintain broad substrate specificity while gaining enhanced thermal stability. The modified enzymes can hydrolyze various phytate forms and remain stable under diverse feed processing conditions, making them universally applicable in different animal feed formulations and processing scenarios

Inventive Principle:
Principle #6Universality (Multi-functionality)

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 phytase variant effectively liberates phosphorous from phytate, improving nutrient availability for animals, reducing phosphorous pollution, and providing a cost-effective alternative to inorganic phosphate supplements.

Implementation Method 1

Through the action of phytase, phytate is generally hydrolysed to give lower inositol-phosphates and inorganic phosphate

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Data Source

PatentEP3118309B1Buttiauxella SP. phytase variants
Publication Date: 2020.09.16 DANISCO US INC
  • EP3118309B1 patent drawingFigure 1~2
  • EP3118309B1 patent drawingFigure 3
  • EP3118309B1 patent drawingFigure 4

AI summary

Provided herein are variants of Buttiauxella sp. phytases that may be used in industrial applications including methods for starch liquefaction, alcohol fermentations and for enhancing phosphate digestion in foods and animal fees.