<figcaptionclassName='caption'>Figure 2. Validation of the feasibility of the metabolic module. (a)Schematic representation of the construction and mechanism of engineered EcN with metabolic module. EcN was transformed with plasmid Ptac-GS via electroporation. After co-culturing with different concentration of NH4Cl for different time, NH3 concentration was measured and calculated by NH3 detection kit based on indigol blue reaction via microplate reader (OD 630nm). The structure of GS was predicted based on AlphaFold3. (b)NH3 concentration after coculturing 0μM, 0.5μM, 5μM and 50μM NH4Cl with engineered EcNs for 12 hours. EcN_vector was transformed with the vector plasmid, pET-32a. Data shows mean±SD, n=3 independent experiments. (c)NH3 concentration in 0h, 4h, 8h, 12h and 24h after coculturing 50μM NH3Cl with engineered EcNs. Data shows mean±SD, n=3 independent experiments.</figcaption>
<h3>Tackling Intrinsic Ammonia of Engineered EcN</h3>
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<h3>Safety concerns of GS enzyme to degrade normal level of ammonia</h3>