Colicin M Antimicrobial Protein for EHEC Contamination Control

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Solution Overview

Problem

Current methods for preventing or reducing contamination with Enterohaemorrhagic Escherichia coli (EHEC) are often specific to particular serotypes, requiring prior knowledge and are not always effective, as they may involve heating, irradiation, or have undesirable effects on food, and existing colicins have narrow target cell specificity.

Innovation Solution

The use of colicin M, which has broad target cell specificity and peptidoglycanase activity, allowing for effective reduction of EHEC contamination across various serotypes without prior serotype identification, expressed in plants using viral vectors for broad antimicrobial activity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If heating or irradiation is used to eliminate EHEC from foods, then EHEC is effectively eliminated, but the food quality is altered in undesirable ways

Engineering Contradiction:
ImproveEHEC elimination effectivenessVSAvoidfood quality alteration
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent replaces physical/thermal methods (heating, irradiation) with a biological method using colicin M protein. Instead of applying external energy to kill bacteria, the invention uses a specifically designed peptide that selectively targets and destroys EHEC cells through membrane disruption, thereby eliminating the need for heating or irradiation that would alter food quality.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The colicin M protein acts as an intermediary substance that mediates between the desired outcome (EHEC elimination) and the constraint (food quality preservation). The protein specifically binds to and disrupts EHEC cell membranes without requiring thermal or radiative energy transfer to the food, thus achieving bacterial elimination while maintaining food integrity.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Reliability

If colicins with narrow target cell specificity are used, then they are effective against specific EHEC strains, but they require prior serotype identification which reduces productivity

Engineering Contradiction:
Improvestrain-specific effectivenessVSAvoidresponse time and processing efficiency
Core Design Contradiction:
ReliabilityVSProductivity

Solution Approach 1:

The patent employs colicin M, which has been engineered or selected to possess broad-spectrum activity against multiple EHEC serotypes including O157:H7, O26, O45, O103, O111, O121, and O145. This single colicin variant can target and eliminate diverse EHEC strains without requiring prior identification of the specific serotype present, thereby providing universal protection across the 'Big 7' pathogenic serogroups.

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

3Adaptability or versatility

If broad-spectrum antimicrobial treatments are applied, then they cover multiple EHEC serotypes, but they may affect beneficial bacteria and have non-specific effects

Engineering Contradiction:
Improvecoverage of EHEC serotypesVSAvoidnon-specific effects on beneficial bacteria
Core Design Contradiction:
Adaptability or versatilityVSObject-affected harmful factors

Solution Approach 1:

The colicin M protein exhibits local quality in its antimicrobial action by specifically targeting Gram-negative bacteria through its unique mechanism of action. The colicin selectively binds to and disrupts the outer membrane and cell wall structures characteristic of Gram-negative bacteria like EHEC, while leaving Gram-positive bacteria and other beneficial microorganisms unaffected due to their different cellular structures.

Inventive Principle:
Principle #3Local quality

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

Colicin M effectively reduces EHEC contamination on food and objects by targeting a wide range of EHEC serotypes, including the 'Big 7' and 'Big 6' groups, with high toxicity and broad spectrum activity, making it suitable for use in food processing and handling without altering the food's quality.

Implementation Method 1

coli M has broad target cell specificity and peptidoglycanase activity, allowing for effective reduction of EHEC contamination

Methodology Applied
Scientific EffectPeptidoglycanase activity: Enzyme

Data Source

PatentEP3097783B1Colicin m for the control of ehec
Publication Date: 2019.11.13 NOMAD BIOSCI
  • EP3097783B1 patent drawingFigure 1A
  • EP3097783B1 patent drawingFigure 1B
  • EP3097783B1 patent drawingFigure 1C

AI summary

The invention provides a method of preventing or reducing contamination of an object such as food with enterohaemorrhagic E. coli (EHEC), comprising contacting said object with colicin M or a derivative thereof.