Engineered Bacteria for Staphylococcus aureus Quorum Sensing Inhibition

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Solution Overview

Problem

Current methods lack effective solutions to inhibit the accessory gene regulator (agr) quorum sensing system in Staphylococcus aureus, which is crucial for controlling virulence factors in pathogenic bacteria, as existing approaches do not efficiently antagonize the system in mixed-microbial environments.

Innovation Solution

Engineered bacterial cells are developed to produce autoinducing peptides (AIP) or non-native AIP analogs by transforming them with specific plasmids encoding agrBD loci, mroQ gene, argB gene, and argD gene, along with promoters, to inhibit the agr quorum sensing system in Staphylococcus aureus.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If existing approaches are used to inhibit the agr quorum sensing system, then some level of inhibition may be achieved, but the effectiveness is insufficient in mixed-microbial environments

Engineering Contradiction:

Improveeffectiveness of agr inhibitionVSAvoidperformance in mixed-microbial environments

Core Design Contradiction:

ReliabilityVSAdaptability or versatility

Solution Approach 1:

The patent modifies the chemical structure of autoinducing peptides by changing parameters such as amino acid sequences, modifications at specific positions (e.g., D5A, D4A mutations), and structural analogs. These parameter changes create non-native AIP analogs that selectively inhibit agr systems across different S. aureus strains and species, achieving reliable inhibition that works effectively in mixed-microbial environments where natural AIPs fail to cross-species barriers.

Inventive Principle:

Principle #35Parameter changes

2Productivity

If native AIP biosynthesis system is manipulated via AgrB/D, then AIP or AIP analogs can be produced, but the system complexity increases due to plasmid transformation and gene expression requirements

Engineering Contradiction:

Improveproduction of AIP or AIP analogsVSAvoidplasmid transformation and gene expression system

Core Design Contradiction:

ProductivityVSDevice complexity

Solution Approach 1:

The patent divides the AIP production system into separate functional modules: (1) plasmid-based expression systems containing specific gene combinations (agrBD loci, mroQ, argB, argD), (2) engineered bacterial host cells, and (3) secretion/transport mechanisms. This segmentation allows independent optimization of each component and simplifies the overall system by using modular plasmids that can be transformed into different host strains depending on the specific AIP analog needed.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent employs plasmids as intermediary carriers that facilitate the introduction and expression of foreign genes (agrBD loci, mroQ, argB, argD) into bacterial cells. These plasmids act as mediators between the desired AIP production function and the host cell machinery, enabling controlled production of native and non-native AIP analogs through regulated gene expression from the plasmid-encoded promoters.

Inventive Principle:

Principle #24Intermediary (Mediator)

Data Source

PatentUS20240352402A1Engineered bacteria for biosynthesis of peptide-based quorum sensing inhibitors

Publication Date: 2024.10.24 WISCONSIN ALUMNI RES FOUND

AI summary

Described herein are engineered bacterial cells capable of producing an autoinducing peptide (AIP) or non-native AIP analog (including AIP inhibitors), methods for producing the AIP or non-native AIP analog, plasmids and kits. The bacterial cells are transformed with at least one plasmid into gram-positive bacterial cells expressing a mutation in the argD gene manipulating the AIP biosynthesis system via the AgrB/D to produce inhibitors of the S. aureus arg quorum sensing.