Engineered EcN Probiotics for AmTARS Secretion in Gut Inflammation
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Solution Overview
Problem
Akkermansia muciniphila, a beneficial gut microbiome bacterium, is difficult to culture and proliferate in vitro, limiting its use for preventing or treating inflammatory diseases, and existing probiotics like Escherichia coli Nissle 1917 (EcN) do not effectively produce and secrete anti-inflammatory TARS (AmTARS) for therapeutic purposes.
Innovation Solution
A transformed Escherichia coli Nissle 1917 (EcN) strain is developed using a vector comprising a nucleic acid encoding TARS of Akkermansia muciniphila (AmTARS), utilizing a 5′-homology arm, a promoter, a ribosome binding site, and a 3′-homology arm, along with a guide RNA and a Cas protein, to stably produce and secrete AmTARS.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If Akkermansia muciniphila is used to produce anti-inflammatory TARS, then therapeutic efficacy is improved, but cultivation difficulty increases
Solution Approach 1:
The patent uses E. coli Nissle 1917 as a host organism to copy and produce the AmTARS protein that would otherwise require cultivation of Akkermansia muciniphila. The host strain contains a plasmid vector with the AmTARS gene, enabling production of the therapeutic protein without needing to cultivate the difficult-to-grow Akkermansia bacterium.
Solution Approach 2:
The patent introduces an intermediary system consisting of a plasmid vector and host strain (E. coli Nissle 1917) that mediates the production of AmTARS. This intermediary system bridges the gap between the desired therapeutic protein and the difficult-to-cultivate source organism, allowing indirect production through a more manageable host.
2Object-affected harmful factors
If E. coli Nissle 1917 is used as a probiotic strain, then safety is improved, but production of anti-inflammatory TARS deteriorates
Solution Approach 1:
The patent merges the safe characteristics of E. coli Nissle 1917 with the therapeutic capability of producing AmTARS by introducing a plasmid vector containing the AmTARS gene into the host strain. This combination creates a probiotic strain that is both safe (inheriting E. coli Nissle 1917 safety) and therapeutically active (capable of producing AmTARS).
Solution Approach 2:
The engineered E. coli Nissle 1917 strain achieves multi-functionality by simultaneously serving as a safe probiotic organism and as a production system for anti-inflammatory TARS. The host strain performs both its original function as a beneficial gut bacterium and the new function of producing therapeutic protein through the introduced plasmid.
3Productivity
If a plasmid vector is introduced into E. coli Nissle 1917 to express AmTARS, then production capability is improved, but strain complexity increases
Solution Approach 1:
The patent extracts and isolates the AmTARS gene from Akkermansia muciniphila and places it into a separate plasmid vector system. This extraction allows the gene to be independently manipulated, optimized for expression, and then introduced into the host strain, thereby managing complexity through modular separation of functional elements.
Data Source
AI summary
The present invention relates to: a transformed strain expressing and secreting Akkermansia muciniphila TARS (AmTARS); and a vector for producing same. More specifically, the present invention relates to an Escherichia coli Nissle 1917 (EcN) strain expressing AmTARS, a composition comprising same, a novel vector for producing same, and an EcN transformation method using same.