Whey Protease Hydrolysis for ACE Inhibitory Peptides

Resolve Bottlenecks,
Find Innovative Solutions
Generate Solutions

Solution Overview

Problem

Current methods for obtaining angiotensin converting enzyme (ACE) inhibitory peptides from whey proteins are inefficient in terms of enzyme consumption and optimization, often resulting in high costs and bitter taste due to hydrophobic amino acid residues, and have not adequately optimized enzyme mixtures to reduce necessary enzyme concentrations for desired IC50 values and inhibition percentages.

Innovation Solution

An optimized method combining two proteases with complementary action and specificity for whey hydrolysis, using a statistical mathematical algorithm to determine optimal enzyme concentrations and ratios, reducing enzyme use while achieving high ACE inhibition percentages and IC50 values, and employing ultrafiltration and HPLC for peptide purification.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If conventional enzymatic hydrolysis methods are used to obtain ACE inhibitory peptides from whey proteins, then peptide production is achieved, but enzyme consumption is high and costs increase

Engineering Contradiction:
Improveenzyme consumptionVSAvoidproduction cost
Core Design Contradiction:
Quantity of substanceVSEase of manufacture

Solution Approach 1:

The patent combines two different proteases (microbial and animal) in a sequential hydrolysis process. The first protease performs initial hydrolysis under specific conditions, followed by a second protease that completes the peptide release. This synergistic combination achieves higher ACE inhibition percentages with lower total enzyme consumption compared to using single enzymes or conventional methods.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The patent optimizes hydrolysis parameters including pH (adjusted to specific ranges for each enzyme), temperature, and hydrolysis time to maximize peptide release efficiency. By controlling these parameters precisely, the process achieves high ACE inhibition with reduced enzyme requirements, directly addressing the contradiction between enzyme quantity and production cost.

Inventive Principle:
Principle #35Parameter changes

2Reliability

If high enzyme concentrations are used to achieve desired ACE inhibition percentages, then inhibition effectiveness is improved, but production costs increase

Engineering Contradiction:
ImproveACE inhibition effectivenessVSAvoidproduction cost
Core Design Contradiction:
ReliabilityVSEase of manufacture

Solution Approach 1:

The sequential use of two proteases with different specificities allows the process to achieve high ACE inhibition percentages (greater than 70%) with lower total enzyme concentrations. The first protease generates intermediate peptides, and the second protease further hydrolyzes them to release active ACE inhibitory peptides, providing cost-effective inhibition effectiveness.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The patent employs continuous hydrolysis where the first protease acts on whey proteins to generate peptides, which are then immediately subjected to further hydrolysis by the second protease. This continuous action ensures complete release of ACE inhibitory peptides without requiring excessive enzyme concentrations, maintaining reliability while reducing cost.

Inventive Principle:
Principle #20Continuity of useful action

3Quantity of substance

If enzymatic hydrolysis is performed to release bioactive peptides from whey proteins, then ACE inhibitory peptides are produced, but bitter taste is generated due to hydrophobic amino acid residues

Engineering Contradiction:
Improvepeptide productionVSAvoidbitter taste
Core Design Contradiction:
Quantity of substanceVSObject-affected harmful factors

Solution Approach 1:

The patent carefully controls hydrolysis parameters (pH, temperature, time) to achieve optimal peptide release while minimizing the generation of hydrophobic peptides responsible for bitter taste. By adjusting these parameters within specific ranges, the process produces ACE inhibitory peptides with reduced bitterness, balancing peptide production with sensory quality.

Inventive Principle:
Principle #35Parameter changes

4Device complexity

If conventional single-enzyme hydrolysis is used, then process simplicity is maintained, but enzyme consumption and cost are high

Engineering Contradiction:
Improveprocess complexityVSAvoidenzyme consumption
Core Design Contradiction:
Device complexityVSQuantity of substance

Solution Approach 1:

The patent integrates two protease treatments into a unified sequential process with optimized parameters for each stage. While the process involves two enzymes, the systematic optimization of pH, temperature, and timing for each hydrolysis stage creates a manageable workflow that achieves superior enzyme efficiency and reduced consumption compared to conventional single-enzyme methods.

Inventive Principle:
Principle #5Merging (Combining)

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 significantly reduces enzyme consumption, achieving ACE inhibition percentages greater than 70% and IC50 values between 30 to 100 µg/ml, with the potential for cost-effective scaling and production of functional foods with high-purity ACE inhibitory peptides.

Implementation Method 1

enzymatic hydrolysis of this whey concentrate for a minimum time of 30 minutes

Methodology Applied
Scientific EffectEnzymatic hydrolysis: Hydrolysis

Implementation Method 2

selecting and adding a mixture having a suitable proportion and amount of two microbial proteases and having complementary action to said whey concentrate for the hydrolysis thereof

Methodology Applied
Scientific EffectProtease action: Enzyme

Implementation Method 3

protein hydrolysate ultrafiltration and ACE inhibitory peptide separation and production

Methodology Applied
Scientific EffectUltrafiltration: Semipermeable Membrane

Data Source

PatentEP2728011B1Optimised method for obtaining angiotensin converting enzyme inhibitory peptides, inhibitory peptides and food containing same
Publication Date: 2017.10.25 QUEIZUAR

AI summary

The present invention relates to an optimized method for obtaining ACE activity inhibitory peptides wherein it includes the steps of whey ultrafiltration for obtaining a whey concentrate, enzymatic hydrolysis, hydrolysate ultrafiltration and inhibitory peptide separation, which comprises selecting and adding a mixture having suitable proportion and amount of two microbial proteases and having complementary action to said whey concentrate for the hydrolysis thereof in order to obtain peptides with an ACE inhibition percentage greater than 70% and an IC50 index of 30 to 100 µg/ml.