PEG Adsorption for Non-Denatured Protein Purification

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Solution Overview

Problem

Existing commercial protein extraction processes denature low temperature-denaturing proteins and are hindered by protein-associated colors and odors, making them unsuitable for food replica products like cheese or meat replicas.

Innovation Solution

A method involving the use of hydrophilic polymers like polyethylene glycol (PEG) to adsorb color and odor compounds from protein solutions, followed by aqueous two-phase separation and ultrafiltration to purify proteins such as rubisco, maintaining their non-denatured state and removing impurities.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If existing commercial protein extraction processes are used, then protein extraction is achieved, but the proteins denature and lose their functional state

Engineering Contradiction:
Improveprotein structural integrityVSAvoidprotein functional state
Core Design Contradiction:
Manufacturing precisionVSReliability

Solution Approach 1:

The patent employs aqueous two-phase extraction using PEG and salt systems, adjusting phase composition ratios and molecular weights to create extraction conditions that maintain protein stability. The process operates at controlled temperatures and pH levels throughout extraction and purification, preventing thermal and chemical denaturation while achieving high purity recovery of functional proteins

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent uses hydrophilic polymers (PEG) and flocculants as intermediary substances that facilitate protein separation and purification without directly contacting or denaturing the protein. These intermediaries form distinct phases that selectively partition proteins while maintaining their native conformation, acting as protective mediators during the extraction process

Inventive Principle:
Principle #24Intermediary (Mediator)

2Productivity

If proteins are extracted using conventional methods, then protein recovery is achieved, but associated colors and odors adversely affect product quality

Engineering Contradiction:
Improveprotein recovery efficiencyVSAvoidcolor and odor impurities
Core Design Contradiction:
ProductivityVSObject-generated harmful factors

Solution Approach 1:

The patent selectively extracts color and odor compounds from protein solutions using adsorbent materials and phase separation techniques. The aqueous two-phase system partitions hydrophobic color/odor molecules into the PEG-rich phase while retaining proteins in the salt-rich phase, effectively removing harmful impurities without sacrificing protein recovery

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent applies different functional zones within the extraction system: one zone optimized for protein solubilization and another for impurity adsorption. By creating local chemical environments with specific pH, ionic strength, and adsorbent concentrations, the process simultaneously achieves high protein recovery and selective impurity removal

Inventive Principle:
Principle #3Local quality

3Manufacturing precision

If multiple purification steps including chromatography are used, then high protein purity is achieved, but process complexity and cost increase

Engineering Contradiction:
Improveprotein purityVSAvoidprocess steps
Core Design Contradiction:
Manufacturing precisionVSDevice complexity

Solution Approach 1:

The patent combines extraction and purification into a single integrated aqueous two-phase system. The PEG-salt phase separation simultaneously achieves protein concentration, purification, and stabilization in one operation, eliminating the need for separate chromatography steps while maintaining high purity and reducing process complexity

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The patent employs a universal PEG-salt extraction system that performs multiple functions: protein solubilization, impurity removal, concentration, and stabilization. This multi-functional approach replaces multiple specialized unit operations with a single versatile process that achieves equivalent or superior results with reduced complexity

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

This method allows for the extraction and purification of high-purity proteins in a single step without chromatography, suitable for scaling up and suitable for use in food replica products, ensuring proteins remain undenatured and free from colors and odors.

Implementation Method 1

hydrophilic polymers, and particularly polyethylene glycol (PEG), can adsorb color and odorous compounds from protein solutions

Methodology Applied
Scientific EffectAdsorption: Adsorption

Implementation Method 2

adding salt to form a two-phase mixture; separating the two-phase mixture to generate a PEG phase and a product phase

Methodology Applied
Scientific EffectLiquid-Liquid Extraction: Liquid-Liquid Extraction

Implementation Method 3

concentrating and diafiltering the filtered product phase to generate a product concentrate. The diafiltering can include using an ultrafiltration membrane system

Methodology Applied
Scientific EffectUltrafiltration: Semipermeable Membrane

Data Source

PatentEP3201336B1Methods for extracting and purifying non-denatured proteins
Publication Date: 2021.09.22 IMPOSSIBLE FOODS INC
  • EP3201336B1 patent drawingFigure 1
  • EP3201336B1 patent drawingFigure 2A
  • EP3201336B1 patent drawingFigure 2B

AI summary

Materials and methods for extracting and purifying proteins are provided. For example, the materials and methods provided herein can be used for extracting and purifying proteins that denature at low temperature.