Comprehensive support for E. coli Nissle 1917 (EcN): authenticated strain access, identity confirmation, fermentation process development, stabilization and formulation, and mechanism-aligned in vitro evaluation. Packages span feasibility to pilot scale with QC gates, enabling EcN programs to benchmark colonization-adjacent traits and microcin/siderophore-associated competitive effects with consistent, comparable data.
Engineering for E. coli includes CRISPR/recombineering, promoter/operon tuning, vector selection, auxotrophy/safety-switch design, and chassis fitness optimization. We couple genotype to phenotype via multiplex growth and expression profiling, mapping burden, copy-number effects, and stress responses to measurable outputs that guide construct pruning and scale-up readiness across E. coli backgrounds.
For engineered E. coli, we quantify segregational and structural stability across serial passages with and without selection pressure, track copy number and expression, and screen for off-target events via WGS/targeted panels. Results pinpoint circuit fragilities, plasmid addiction needs, or integration-site effects to de-risk downstream fermentation and host-interaction experiments.
We build mechanism-relevant assays for E. coli: competitive exclusion against pathobionts, siderophore/microcin-linked effects, metabolite outputs, and barrier integrity or cytokine shifts. Data packages include orthogonal readouts (e.g., CFU dynamics, TEER, reporter panels) to anchor claims in quantifiable biology and enable apples-to-apples comparisons across variants.
Using epithelial monolayers (Caco-2/HT29-MTX), primary/organoid-derived intestinal models, and immune-competent co-cultures, we assess E. coli adhesion/colonization, tight junction markers, mucus interactions, and host response signatures. These models provide barrier-relevant endpoints without overfitting to a single cell line, supporting iterative design and formulation refinement.
We translate E. coli bench runs to robust fed-batch/continuous regimes by optimizing carbon feed, DO, pH, and temperature, then scale by transfer-ready criteria (e.g., kLa, oxygen uptake rate). Deliverables balance biomass/viability with expression targets and link upstream parameters to downstream stability, drying, and packaging needs.
For E. coli lines, we screen virulence genes and mobile elements, evaluate prophage/lysogeny risk, and quantify endotoxin and residual DNA per harmonized test chapters. Outputs support internal risk management and communication with oversight bodies in research contexts.
We provide MIC/MLC panels for E. coli using current CLSI methods and breakpoints, complemented by AMR-gene profiling and cross-resistance interpretation. Results contextualize selective markers and environmental exposures, informing strain stewardship and documentation for research governance.
Fig.1 Identification of gene knock-out.
Fig.2 Identification of genes knock-in.
