Interindividual Differences in In Vitro Human Intestinal Microbial Conversion of 3-Acetyl-DON and 15-Acetyl-DON

Abstract

In order to evaluate the potential differences between 3-Ac-DON and 15-Ac-DON in the human intestinal microbial metabolism, human fecal samples were anaerobically cultured in vitro. Quantitative fecal microbiota characteristics were obtained by 16S rRNA sequencing, and the data revealed several genera that may be relevant for the transformation of the acetylated DONs. Significant differences in the level of 3-Ac-DON and 15-Ac-DON conversion were observed among microbiota from different human individuals. 3-Ac-DON could be rapidly hydrolyzed; a ten-fold difference was observed between the highest and lowest in vitro conversion after 4 h. However, 15-Ac-DON was not fully transformed in the 4 h culture of all the individual samples. In all cases, the conversion rate of 3-Ac-DON was higher than that of 15-Ac-DON, and the conversion rate of 3-Ac-DON into DON varied from 1.3- to 8.4-fold that of 15-Ac-DON. Based on in vitro conversion rates, it was estimated that 45–452 min is required to convert all 3-Ac-DON to DON, implying that deacetylation of 3-Ac-DON is likely to occur completely in all human individuals during intestinal transit. However, for conversion of 15-Ac-DON, DON formation was undetectable at 4 h incubation in 8 out of the 25 human samples, while for 7 of these 8 samples conversion to DON was detected at 24 h incubation. The conversion rates obtained for these seven samples indicated that it would take 1925–4805 min to convert all 15-Ac-DON to DON, while the other 17 samples required 173–734 min. From these results it followed that for eight of the 25 individuals, conversion of 15-Ac-DON to DON was estimated to be incomplete during the 1848 min intestinal transit time. The results thus indicate substantial interindividual as well as compound specific differences in the deconjugation of acetylated DONs. A spearman correlation analysis showed a statistically significant relationship between deconjugation of both acetyl-DONs at 4 h and 24 h incubation. Based on the in vitro kinetic parameters and their scaling to the in vivo situation, it was concluded that for a substantial number of human individuals the deconjugation of 15-Ac-DON may not be complete upon intestinal transit.