# Characterization of dietary choline uptake by the gut microbiome reveals choline assimilating microbes and influences on host choline metabolism

**Authors:** Elizabeth Johnson, Paula Bañuelos, Janine Comrie, Annett Richter, Henry Le, Sharon Thompson

PMC · DOI: 10.21203/rs.3.rs-8491198/v1 · Research Square · 2026-01-07

## TL;DR

This study shows how gut bacteria like L. reuteri process dietary choline, affecting host metabolism and lipid production.

## Contribution

The study identifies new microbial pathways for choline assimilation and their impact on host lipid metabolism.

## Key findings

- L. reuteri assimilates choline and converts it to long chain fatty acylcholines.
- L. reuteri colonization increases serum choline and lysophosphatidylcholine production.
- Choline assimilation by L. reuteri upregulates hepatic genes involved in lipid metabolism.

## Abstract

Choline is an essential nutrient with diverse roles in host metabolism; however, the current understanding of its microbial fate is largely restricted to trimethylamine production. Here, we apply the BioOrthogonal-labeling, Sorting, Sequencing, and mass Spectrometry (BOSSS) workflow to map dietary choline-specific gut microbial interactions. Using an alkyne-modified choline analog (propargylcholine) in mice, we fluorescently label and flow-sort choline-assimilating gut bacteria, identifying a varied set of taxa enriched in probiotic species, including Limosilactobacillus reuteri (L. reuteri). In vitro studies confirm that L. reuteri assimilates choline and converts it to long chain fatty acylcholines. Metabolomic and transcriptomic profiling revealed that L. reuteri colonization, with a choline sufficient diet, selectively elevates serum choline and increases lysophosphatidylcholine production, findings corroborated by transcriptomic evidence of upregulated hepatic genes involved in lipid metabolism. These findings uncover a new metabolic fate for dietary choline, expand the known repertoire of microbiome-derived lipids, and illustrate how specific host–microbe–diet interactions can influence host status.

## Linked entities

- **Chemicals:** choline (PubChem CID 305), trimethylamine (PubChem CID 1146), lysophosphatidylcholine (PubChem CID 5311264)
- **Species:** Limosilactobacillus reuteri (taxon 1598), Mus musculus (taxon 10090)

## Full-text entities

- **Genes:** ALB (albumin) [NCBI Gene 280717], CUTC (cutC copper transporter) [NCBI Gene 51076] {aka CGI-32}
- **Diseases:** inflammatory (MESH:D007249), neurodegenerative disorders (MESH:D019636), muscle atrophy (MESH:D009133), MCD (MESH:D002796), liver disease (MESH:D008107), sterility (MESH:D007246)
- **Chemicals:** formic acid (MESH:C030544), Cu(I) (MESH:C073870), acetylcholine (MESH:D000109), alkyne (MESH:D000480), azide (MESH:D001386), C18:0 (MESH:C031183), AF647 (MESH:C569686), TMA (MESH:C023336), palmitoylcholine (MESH:C032703), glycerol (MESH:D005990), isopentane (MESH:C067038), DAPI (MESH:C007293), methanol (MESH:D000432), triethylamine (MESH:C016162), H&amp;E (MESH:D006371), palmitoyl chloride (MESH:C032023), betaine (MESH:D001622), ice (MESH:D007053), NaCl (MESH:D012965), B (MESH:D001895), water (MESH:D014867), phosphatidylcholine (MESH:D010713), copper (MESH:D003300), sodium acetate (MESH:D019346), PC (MESH:C000603143), phospholipids (MESH:D010743), fatty acid (MESH:D005227), glycogen (MESH:D006003), sphingomyelin (MESH:D013109), TRIzol (MESH:C411644), TE (MESH:D013691), isoamyl alcohol (MESH:C029683), chloroform (MESH:D002725), isopropanol (MESH:D019840), Lipids (MESH:D008055), C19:1-PC (-), methionine (MESH:D008715), Triton X-100 (MESH:D017830), acetonitrile (MESH:C032159), sphingolipid (MESH:D013107), copper(II) sulfate (MESH:D019327), LPC (MESH:D008244), SDS (MESH:D012967), MDC (MESH:C039696), nitrogen (MESH:D009584), polystyrene (MESH:D011137), cholesterol (MESH:D002784), propargyl bromide (MESH:C458239), EtOH (MESH:D000431), phenol (MESH:D019800), Choline (MESH:D002794), formalin (MESH:D005557)
- **Species:** Desulfovibrio desulfuricans (species) [taxon 876], Mus musculus (house mouse, species) [taxon 10090], Turicibacter (genus) [taxon 191303], Bacteria Latreille et al. 1825 (Bacteria stick insect, genus) [taxon 629395], Oryza sativa (Asian cultivated rice, species) [taxon 4530], gut metagenome (species) [taxon 749906], Klebsiella pasteurii (species) [taxon 2587529], Escherichia fergusonii (species) [taxon 564], Bifidobacterium pseudolongum (species) [taxon 1694], Lactobacillus intestinalis (species) [taxon 151781], Romboutsia ilealis (species) [taxon 1115758], Eukaryota (eukaryotes, domain) [taxon 2759], Lactobacillus johnsonii (species) [taxon 33959], Lentinula sp. R (species) [taxon 1445722], Limosilactobacillus reuteri (species) [taxon 1598], Homo sapiens (human, species) [taxon 9606], Bacteroides intestinalis (species) [taxon 329854]
- **Mutations:** C032825G
- **Cell lines:** AF647 — Homo sapiens (Human), Ataxia telangiectasia syndrome, Finite cell line (CVCL_1L40)

## Full text

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## Figures

4 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12803358/full.md

## References

42 references — full list in the complete paper: https://tomesphere.com/paper/PMC12803358/full.md

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Source: https://tomesphere.com/paper/PMC12803358