High-throughput screening platform for identifying postbiotics that reduce trimethylamine production

Overview

This solution is a high-throughput screening platform designed to identify postbiotics from fermented plant foods that inhibit the gut microbial conversion of choline to trimethylamine (TMA). TMA is a deleterious microbial metabolite associated with inflammatory bowel disease and cardiovascular disease, as it is subsequently oxidized to trimethylamine N-oxide (TMAO) in the liver. Unlike conventional microbiome sequencing approaches that identify broad population shifts without functional context, this platform targets a specific, functionally meaningful outcome: reducing TMA production. By screening postbiotic candidates against a validated anaerobic fermentation model, the platform enables rapid identification of bioactive food-derived compounds with measurable health-relevant effects.

Technical specifications

Validated screening workflow:

• Uses an established high-throughput 96-well anaerobic human fecal fermentation model
• Employs isotopically labeled substrate (choline-d9) to eliminate background interference, producing TMA-d9 as the measurable output
• Validated stoichiometry showing 1:1 relationship between choline utilization and TMA production, dependent on fecal inoculum presence
• Rapid 96-well UPLC-MS/MS method for quantifying choline-d9 consumption and TMA-d9 production
• Includes simulated gastric and small intestinal digestion prior to fermentation
• Measures bacterial viability via MTT assay to distinguish cytostatic or cytotoxic effects from true TMA inhibition
• Uses unfermented foods and blank digesta as controls
• Calculates areas-under-the-curve and kinetic parameters for both analytes

Demonstrated efficacy with reference compounds:

• Dietary phytochemicals including chlorogenic acid, gallic acid, and epicatechin showed inhibitory activity
• Gut microbial metabolites including 3-(3,4-dihydroxyphenyl)propionic acid and 3-(3,4-dihydroxyphenyl)acetic acid reduced TMA production
• Whole foods including cocoa, coffee, and artichoke inhibited choline-to-TMA conversion

Technology readiness level

The platform has been extensively validated with reference compounds and whole foods, confirming reproducibility and specificity. The screening methodology is established and ready for application to novel fermented food samples. Future validation will apply this platform to evaluate fermented foods from industry partners, measuring their potential to inhibit choline conversion to TMA by fecal microbiota. The approach is positioned for pilot-scale screening campaigns and collaborative evaluation of proprietary fermented food products.