Development of a High-Throughput Method to Study the Inhibitory Effect of Phytochemicals on Trimethylamine Formation |
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Authors: | Lisard Iglesias-Carres Lauren A. Essenmacher Kathryn C. Racine Andrew P. Neilson |
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Affiliation: | 1.Plants for Human Health Institute, Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Kannapolis, NC 28081, USA; (L.I.-C.); (K.C.R.);2.Department of Food Science and Technology, Virginia Polytechnic and State University, Blacksburg, VA 24061, USA; |
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Abstract: | Choline is metabolized by the gut microbiota into trimethylamine (TMA), the precursor of pro-atherosclerotic molecule trimethylamine N-oxide (TMAO). A reduction in TMA formation has shown cardioprotective effects, and some phytochemicals may reduce TMA formation. This study aimed to develop an optimized, high-throughput anaerobic fermentation methodology to study the inhibition of choline microbial metabolism into TMA by phenolic compounds with healthy human fecal starter. Optimal fermentation conditions were: 20% fecal slurry (1:10 in PBS), 100 µM choline, and 12 h fermentation. Additionally, 10 mM of 3,3-dimethyl-1-butanol (DMB) was defined as a positive TMA production inhibitor, achieving a ~50% reduction in TMA production. Gallic acid and chlorogenic acid reported higher TMA inhibitory potential (maximum of 80–90% TMA production inhibition), with IC50 around 5 mM. Neither DMB nor gallic acid or chlorogenic acid reduced TMA production through cytotoxic effects, indicating mechanisms such as altered TMA-lyase activity or expression. |
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Keywords: | atherosclerosis gallic acid chlorogenic acid microbiota trimethylamine |
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