Variant Phytase Enzyme Stability in Feed Processing

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Solution Overview

Problem

Phytase enzymes in animal feeds are not stable under high temperatures and acidic conditions, leading to inefficient phytate degradation and increased costs due to the need for external phosphorus supplementation, which also contributes to environmental pollution.

Innovation Solution

Development of variant phytase enzymes with specific amino acid substitutions, such as I55V/G157Q/R159Y/Y255D/F354Y, and additional substitutions like H60Q/D69N/K74D/S120R/N137P, enhancing thermostability and pH stability, thereby maintaining activity during feed processing and digestion.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If phytase is added to animal feeds to degrade phytate, then phosphorus availability is improved, but the enzyme is inactivated by high temperature during feed processing and low pH in the gastrointestinal tract

Engineering Contradiction:
Improvephytase activity stabilityVSAvoidfeed processing temperature
Core Design Contradiction:
ReliabilityVSTemperature

Solution Approach 1:

The patent applies parameter changes by modifying the amino acid sequence of phytase to alter its thermal and pH stability parameters. Specifically, substitutions at positions 55, 157, 159, 255, 354, and 380 were made to enhance the enzyme's stability under feed processing conditions (65-85°C) and gastrointestinal pH conditions, allowing it to maintain activity where wild-type phytase would be inactivated

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent creates a copy of the phytase enzyme with modified amino acid sequences based on sequences from thermally stable sources. By copying and adapting sequences from thermophilic organisms, the invention produces a variant phytase that inherits thermal stability properties while maintaining phytate degradation function

Inventive Principle:
Principle #26Copying

2Productivity

If wild-type phytase is used in feed, then phosphorus liberation is achieved, but the enzyme loses activity under acidic conditions and protease degradation

Engineering Contradiction:
Improvephytate degradation efficiencyVSAvoidacidic conditions and protease degradation
Core Design Contradiction:
ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent modifies the enzyme's resistance parameters to acidic conditions and protease degradation through specific amino acid substitutions. The variant phytase maintains catalytic activity at low pH levels found in the gastrointestinal tract and resists proteolytic degradation, thereby preserving productivity in the harmful environment of the animal digestive system

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

Instead of using expensive inorganic phosphorus supplements that provide only temporary nutritional value, the patent employs a engineered phytase enzyme that continuously degrades phytate in the digestive tract, providing sustained phosphorus liberation without the need for repeated supplementation

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

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 variant phytase enzymes exhibit improved thermostability and pH stability, ensuring effective phytate degradation and reducing phosphorus excretion, thus enhancing nutritional value and reducing environmental impact.

Implementation Method 1

Phytase is a phosphatase that catalyzes the hydrolysis of O-P bonds in phytate and releases inorganic usable phosphorous

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Data Source

PatentEP3263698B1Novel phytases and uses thereof
Publication Date: 2021.03.24 FORNIA BIOSOLUTIONS INC
  • EP3263698B1 patent drawingFigure 1
  • EP3263698B1 patent drawingFigure 2
  • EP3263698B1 patent drawingFigure 3

AI summary

The present invention relates to variant phytase enzymes and their use thereof.