PlyP825 Endolysin Modular Domain Segmentation for Listeria Control
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Solution Overview
Problem
Current methods for controlling Listeria contamination and infection are limited by the lack of effective, broad-host-range bacteriophages that can specifically target and lyse Listeria serovars, particularly serovar 3, which is clinically relevant, and existing endolysins face challenges with proteolytic instability and limited spectrum of activity.
Innovation Solution
A novel Listeria bacteriophage, ProCC P825, and its encoded endolysin, PlyP825, are developed, capable of lysing Listeria serovars 1/2, 3, 4, 5, and 6, with a unique ability to target serovar 3, and exhibit improved stability and broad-spectrum activity through a modular organization of enzymatically active and cell wall binding domains.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing endolysins are used to control Listeria, then Listeria contamination can be reduced, but the endolysins suffer from proteolytic instability and limited spectrum of activity
Solution Approach 1:
The endolysin PlyP825 is segmented into distinct functional domains: an N-terminal catalytic domain (residues 1-162) responsible for lytic activity and a C-terminal cell wall binding domain (residues 163-315) responsible for substrate recognition. This modular organization allows the enzyme to maintain stability through its structured domains while achieving broad spectrum activity through the specific properties of the cell wall binding domain that recognizes conserved structures across multiple Listeria serovars.
2Measurement precision
If bacteriophages are used to specifically target Listeria serovars, then specific killing of bacteria is achieved, but the host-specificity limits the range of strains that can be controlled
Solution Approach 1:
The endolysin PlyP825 achieves universality by targeting conserved cell wall structures present across multiple Listeria serovars (1/2, 3, 4, 5, and 6) rather than serovar-specific antigens. The C-terminal cell wall binding domain recognizes conserved peptidoglycan structures that are universally present in Listeria species, enabling a single enzyme to function against diverse strains while maintaining specific killing activity.
3Adaptability or versatility
If broad-spectrum endolysins are developed to target multiple Listeria serovars, then the spectrum of activity is improved, but the complexity of the enzyme structure increases
Solution Approach 1:
The enzyme is organized into two compact, independently folded domains with clear functional separation. The N-terminal catalytic domain (162 residues) and C-terminal binding domain (153 residues) are connected by a flexible linker, allowing each domain to fold independently and perform its specific function. This modular segmentation achieves broad spectrum activity through the binding domain's recognition of conserved structures while keeping the overall structure manageable and suitable for production.
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
PlyP825 effectively reduces Listeria titers, demonstrating consistent lytic activity across various strains and environments, including milk, with enhanced stability compared to existing endolysins, making it suitable for controlling Listeria contamination and infection.
Implementation Method 1
Endolysins from Listeria bacteriophages are promising tools for detection and control of Listeria contamination and infection. These proteins have a modular organization, which is characterized by an N-terminal localized enzymatically active domain (EAD), which contributes lytic activity
Data Source
AI summary
The present invention relates to a novel Listeria bacteriophage designated ProCC P825. In particular, the present invention relates to the endolysin PlyP825 encoded by the novel phage ProCC P825 and uses of the novel endolysin PlyP825 for controlling Listeria contamination and infection.