Engineered E. coli Constitutive Microcin Production Against Enteric Bacteria
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Solution Overview
Problem
The increasing prevalence of drug-resistant bacteria, particularly carbapenem-resistant Enterobacteriaceae, poses a significant threat due to high morbidity, mortality, and healthcare costs, with limited treatment options, necessitating the development of novel therapeutics to selectively target and eliminate these pathogens from the gastrointestinal tract.
Innovation Solution
Genetically engineered microorganisms, such as E. coli Nissle 1917, are designed to constitutively produce microcins like MccH47 and MccI47 using a self-retaining multicopy plasmid vector, allowing continuous expression and secretion of these antimicrobial peptides to target and kill drug-resistant bacteria like Klebsiella pneumoniae, without the need for antibiotic selection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional antibiotics are used to treat drug-resistant bacteria, then treatment options are limited, but antibiotic resistance increases and healthcare costs increase
Solution Approach 1:
The patent uses bacteriocins produced by engineered E. coli Nissle 1917 to convert the harmful effect of bacterial competition into a beneficial therapeutic effect. The engineered bacteria produce microcins that selectively kill drug-resistant Enterobacteriaceae while the probiotic E. coli Nissle 1917 provides beneficial gut colonization effects, thus converting the potential harm of bacterial competition into a dual benefit of pathogen elimination and probiotic support
Solution Approach 2:
The patent introduces an intermediary organism (engineered E. coli Nissle 1917) that produces bacteriocins to mediate the elimination of drug-resistant bacteria. Instead of using antibiotics directly, the engineered probiotic bacteria serve as living factories that produce and secrete microcins, which then act as the actual antimicrobial agents against the target pathogens
2Adaptability or versatility
If engineered microorganisms are used to produce microcins, then selective targeting of drug-resistant bacteria is achieved, but device complexity increases
Solution Approach 1:
The patent uses the universal probiotic strain E. coli Nissle 1917 that can serve multiple functions: (1) constitutive colonization of the gastrointestinal tract, (2) production of bacteriocins against drug-resistant bacteria, and (3) provision of probiotic benefits. This multi-functionality reduces the need for multiple separate therapeutic agents while maintaining selective targeting capability
Solution Approach 2:
The engineered E. coli Nissle 1917 is designed to be self-sufficient by incorporating constitutive promoters that enable continuous production of microcins without requiring external induction or complex regulatory systems. The bacteria autonomously produce and secrete the antimicrobial peptides while maintaining their probiotic functions
3Productivity
If constitutive production of microcins is implemented, then continuous elimination of drug-resistant bacteria is achieved, but loss of substance increases due to continuous peptide production
Solution Approach 1:
The patent employs constitutive production of microcins at low to moderate levels rather than maximizing production intensity. The constitutive promoters drive continuous but controlled expression of bacteriocin genes, providing sufficient antimicrobial activity for pathogen elimination while avoiding excessive resource consumption and metabolic burden on the engineered bacteria
4Reliability
If engineered E. coli Nissle 1917 is administered, then gastrointestinal colonization of drug-resistant bacteria is reduced, but ease of operation decreases due to requiring genetic engineering
Solution Approach 1:
The patent performs preliminary genetic engineering of E. coli Nissle 1917 in controlled laboratory settings before clinical administration. The constitutive microcin production capability is pre-installed in the bacteria through genetic modification, allowing the engineered strain to be manufactured once and then administered repeatedly without requiring further genetic manipulation at the point of use
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
The engineered microorganisms effectively reduce gastrointestinal colonization of drug-resistant bacteria, including hypervirulent strains, thereby decreasing host-to-host transmission and minimizing systemic side effects, while maintaining stability in the gut microbiota.
Implementation Method 1
The constitutive promoter allows for continual transcription and expression of the one or more microcin genes
Implementation Method 2
The constitutive promoter drives the expression of the microcin operon constitutively
Implementation Method 3
allowing continuous expression and secretion of these antimicrobial peptides to target and kill drug-resistant bacteria like Klebsiella pneumoniae
Data Source
AI summary
This disclosure relates to genetically engineered microorganisms for treating or reducing the risk of bacterial infections or dysbiosis, and further discloses methods of making and using such microorganisms.