Comprehensive assessment of functional effects of commonly used sweeteners on ex vivo human gut microbiome

Abstract

The gut microbiome composition and function are associated with health and diseases. Sweeten-ers are widely used food additives, although many studies using animal models have linked sweetener consumption to gut microbial changes and health issues. Whether sweeteners directly change the human gut microbiome functionality remains largely unknown. In this study, we sys-tematically investigated the responses of five human gut microbiomes to 21 common sweeteners, using an approach combining high-throughput ex vivo microbiome culturing and metaproteomics to quantify functional changes in different taxa. Hierarchical clustering based on metaproteomic responses of individual microbiomes resulted in two clusters. The first cluster was composed of non-caloric artificial sweeteners (NAS) and two sugar alcohols with shorter carbon backbones (4-5 carbon atoms), and the second cluster was composed of sugar alcohols with longer carbon backbones. The metaproteomic functional responses of the second cluster were similar to the prebiotic fructooligosaccharides and kestose, indicating that these sugar alcohol-type sweeteners have potential prebiotic functions. This study provides a comprehensive evaluation of the direct effects of commonly used sweeteners on the functions of the human gut microbiome using a func-tional metaproteomics approach, improving our understanding of the roles of sweeteners on mi-crobiome-associated human health and disease issues.

An economic and robust TMT labeling approach for high throughput proteomic and metaproteomic analysis

“…Marked differences between the metaproteomes of cultured microbiomes with and without the treatment of kestose were observed along the first principal component (horizontal axis), which explains 49% of the variations displayed in PCA score plot. These results are in agreement with previous studies showing the modulating effects of kestose on in vitro cultured microbiota [24, 25]. As expected, microbiome culturing using fresh stools clustered closer to the baseline samples compared to those using frozen microbiotas.…”