Transgenic Bacterial Host Cell for MEP Pathway Screening
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Solution Overview
Problem
The widespread resistance of bacteria to antibiotics and the environmental toxicity of herbicides necessitate the development of new antibacterial and herbicidal agents that are effective yet less harmful to humans and the environment, with a need for methods to screen such compounds that target the methylerythritol phosphate biosynthetic pathway for essential isoprenoid compounds.
Innovation Solution
A transgenic bacterial host cell with disruptions in the methylerythritol phosphate pathway is used to screen for novel antibacterial and herbicidal agents, where the genome contains a cloned transgene to replace impaired genes, and a mevalonic acid mini operon provides an alternate pathway, allowing for the identification of compounds that inhibit the MEP pathway without toxicity to humans or significant environmental impact.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If new antibiotics are discovered and introduced to treat resistant bacteria, then bacterial resistance is overcome, but the new agents become toxic to humans and kill resident microflora
Solution Approach 1:
The patent targets a specific pathway (MEP pathway for isoprenoid biosynthesis) that is present in bacteria and plants but absent in humans. By designing compounds that selectively inhibit this bacterial-specific pathway, the invention achieves local quality differentiation - the antibacterial agent is effective against bacteria while being non-toxic to human cells, as humans utilize the mevalonate pathway instead
Solution Approach 2:
The invention segments the isoprenoid biosynthesis pathways into two distinct routes: the mevalonate pathway (present in eukaryotes including humans) and the MEP pathway (present in bacteria and plants). By targeting the MEP pathway specifically, the patent creates a segmented approach that allows selective antibacterial activity while sparing human biological systems
2Productivity
If effective herbicides are used to combat undesirable plants, then crop yields increase, but the herbicides cause environmental harm and potential harm to humans
Solution Approach 1:
The patent applies local quality by targeting the MEP pathway which is present in plants and bacteria but absent in animals. Herbicides designed to inhibit this pathway achieve selective plant control while being safe for humans and animals, thereby reducing environmental harm while maintaining crop productivity
3Object-affected harmful factors
If the MEP pathway is targeted for screening antibacterial and herbicidal agents, then selective toxicity is reduced, but a screening system is required to identify effective compounds
Solution Approach 1:
The patent introduces a transgenic bacterial host cell as an intermediary screening system. This host contains disruptions in MEP pathway genes and relies on a cloned transgene for pathway function. The transgenic system acts as a mediator that translates compound activity against the MEP pathway into measurable phenotypic outcomes, simplifying the screening process while maintaining selectivity
Solution Approach 2:
The transgenic bacterial host cell is designed to be self-sufficient for screening purposes. It contains all necessary genetic components (disrupted endogenous genes, cloned transgenes, and the mevalonate mini-operon) within a single organism that can autonomously indicate pathway inhibition through growth phenotypes, eliminating the need for complex external screening apparatus
Data Source
AI summary
A transgenic bacterial host cell that can be used as a screen for novel antibiotics and herbicides is provided. The genome of the transgenic bacterial host cell comprises disruptions in a first endogenous gene in the MEP pathway) and a transgene that functionally replaces the disrupted first gene. In other embodiments, the genome comprises a disruption in a first endogenous gene in the MEP pathway and a second endogenous gene which is located downstream of the first gene in the MEP pathway. A transgene that functionally replaces the disrupted downstream gene is cloned into the host cell. A mini operon containing the essential genes for the MVA pathway may also be cloned into the host cell. The transgenic host cell may be used in a method for screening compounds for antibiotic and herbicidal properties. The agents determined by the screening method may be used to kill bacteria or plants.