DOI: 10.5061/dryad.98sf7m0rt
title: Untargeted metabolomics data from Faecalibacterium prausnitzii FAAH experiment

description: Undernutrition in Bangladeshi children is associated with disruption of
 postnatal gut microbiota assembly; compared to standard therapy, a
 microbiota-directed complementary food (MDCF) significantly improved their
 ponderal and linear growth. Here, we characterize a fatty acid amide
 hydrolase (FAAH) from a growth-associated intestinal strain of
 Faecalibacterium prausnitzii cultured from these children. This enzyme,
 expressed and purified from E. coli, hydrolyzes a variety of N-acylamides,
 including oleoylethanolamide (OEA), neurotransmitters, and quorumsensing
 N-acyl homoserine lactones, and synthesizes a range of N-acylamides,
 notably N-acyl amino acids. Treating germ-free mice with N-oleoylarginine
 and N-oleolyhistidine, major products of FAAH OEA metabolism, markedly
 affected expression of intestinal immune function pathways. Administering
 MDCF to Bangladeshi children significantly reduced fecal OEA, a satiety
 factor whose levels were negatively correlated with the abundance and
 expression of their F. prausnitzii FAAH. This enzyme, structurally and
 catalytically distinct from mammalian FAAH, is positioned to regulate
 levels of a variety of bioactive molecules.
description: Untargeted metabolomic analyses were performed using an Agilent
 1290 LC system coupled to an Agilent Model 6545 Qtof mass spectrometer
 (Santa Clara, CA). Reversed phase separation was achieved on a BEH C18
 column (2.1 x 150 mm, 1.7 μm, Waters Corp., Milford, MA) that was heated
 to 35 °C. For analyses carried out in the positive ESI mode, the mobile
 phase consisted of 0.1% formic in water (A) and 0.1% formic acid in
 acetonitrile (B). For analyses in the negative mode, the mobile phase
 consisted of 5mM ammonium bicarbonate formic in water (A) and 5mM ammonium
 bicarbonate in acetonitrile/water (95:5 v/v) (B). A flow rate of 0.3
 mL/minute was applied (gradient program: from 0 to 14 minutes, mobile
 phase B eluted from 5% to 100%, followed by 3 minutes at 100% of B). An
 equilibration time of 3 minutes was used. Normal phase separation was
 conducted on a Cortex HILIC column (2.1 x 150 mm, 1.6 μm, Waters Corp.,
 Milford, MA) that was heated to 45 °C. 10 mm ammonium acetate in 5/95
 acetonitrile/water (A) and 10 mm ammonium acetate in 95/5
 acetonitrile/water (B) were the mobile phases. Metabolites were eluted
 from the columns at 0.4 ml/min using a 1–60% phase A gradient over 17 min
 followed by 4-minute-long equilibration. Data were collected in the range
 from m/z 50 to 1000, and m/z 50 to 650 for MS full-scan analysis and MS/MS
 analysis, respectively. The key parameters of Qtof were set as follows:
 nozzle voltage, 1000 V for positive and 1500V for negative; capillary
 voltage, 3000 V for positive and 3500v for negative; drying gas,
 N<sub>2</sub>; drying gas flow rate, 10 L/min; collision gas,
 high purity N2; drying gas (N<sub>2</sub>) temperature, 325
 °C; vaporizer/sheath gas temperature, 350 °C; sheath gas flow rate, 12
 L/min. 
description: # Untargeted metabolomics data
 [https://doi.org/10.5061/dryad.98sf7m0rt](https://doi.org/10.5061/dryad.98sf7m0rt) UPLC Qtof untargeted analysis of cecal samples from 4-5-week-old germ-free C57Bl/6J mice colonized with either 13 or 14 genome-sequenced human gut bacterial taxa ## Description of the data and file structure Files are named in the format of experiment.mouse number.group.diet. These files are the original Agilent Qtof raw data files.  Non-proprietary free software such as  ProteoWizard, OpenMS, MS-DIAL, MZmine and MetaboAnalyst can process these files.