Metaproteomic and 16S rRNA Gene Sequencing Analysis of the Infant Fecal Microbiome

International Journal of Molecular Sciences

Aude Tartiere¹  Laetitia Cortes¹  Julie Piquenot¹  Daniel Chelsky¹  Harm Wopereis²  Jan Knol²  JoostW. Gouw²  Sebastian Tims² 

[1] Caprion Biosciences, 201 President Kennedy, Suite PK-3900, Montreal, QC H2X3Y7, Canada;Danone Nutricia Research, Gut Biology and Microbiology, Uppsalalaan 12, 3584 CT Utrecht, The Netherlands;

关键词: metacluster;    metabolism;    infants;    mass spectrometry;    fecal;    intestinal;    microbiome;   

DOI  :  10.3390/ijms20061430

来源: DOAJ

【 摘 要 】

A metaproteomic analysis was conducted on the fecal microbiome of eight infants to characterize global protein and pathway expression. Although mass spectrometry-based proteomics is now a routine tool, analysis of the microbiome presents specific technical challenges, including the complexity and dynamic range of member taxa, the need for well-annotated metagenomic databases, and high inter-protein sequence redundancy and similarity. In this study, an approach was developed for assessment of biological phenotype and metabolic status, as a functional complement to DNA sequence analysis. Fecal samples were prepared and analysed by tandem mass spectrometry and a homology-based meta-clustering strategy was used to combine peptides from multiple species into representative proteins. In total, 15,250 unique peptides were sequenced and assigned to 2154 metaclusters, which were then assigned to pathways and functional groups. Differences were noted in several pathways, consistent with the dominant genera observed in different subjects. Although this study was not powered to draw conclusions from the comparisons, the results obtained demonstrate the applicability of this approach and provide the methods needed for performing semi-quantitative comparisons of human fecal microbiome composition, physiology and metabolism, as well as a more detailed assessment of microbial composition in comparison to 16S rRNA gene sequencing.

【 授权许可】

Unknown