Determination of Butyrate Synthesis Capacity in Gut Microbiota: Quantification of but Gene Abundance by qPCR in Fecal Samples

Daskova, Nikola; Heczkova, Marie; Modos, Istvan; Videnska, Petra; Splichalova, Petra; Pelantova, Helena; Kuzma, Marek; Gojda, Jan; Cahova, Monika

doi:10.3390/biom11091303

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Determination of Butyrate Synthesis Capacity in Gut Microbiota: Quantification of but Gene Abundance by qPCR in Fecal Samples

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Monika Cahova

Institute for Clinical and Experimental Medicine, Videnska 1958, 140 21 Prague 4, Czech Republic

First Faculty of Medicine, Charles University, Katerinska 1660/32, 121 08 Prague, Czech Republic

RECETOX, Faculty of Science Masaryk University, Kamenice 753, 625 00 Brno, Czech Republic

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Institute of Microbiology, AS CR, Videnska 1083, 142 20 Prague 4, Czech Republic

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Department of Analytical Chemistry, Faculty of Science, Palacky University Olomouc, 17. Listopadu 1192/12, 779 00 Olomouc, Czech Republic

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Department of Internal Medicine, Kralovske Vinohrady University Hospital and Third Faculty of Medicine, Charles University, Srobarova 1150, 100 34 Prague 10, Czech Republic

Author to whom correspondence should be addressed.

Academic Editor: Piotr Ceranowicz

Biomolecules 2021, 11(9), 1303; https://doi.org/10.3390/biom11091303

Received: 28 July 2021 / Revised: 30 August 2021 / Accepted: 1 September 2021 / Published: 2 September 2021

(This article belongs to the Special Issue Nutrition, Gut Microbiome/Metabolome and Immune System: Prospects for Non-Communicable Diseases)

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Abstract

Butyrate is formed in the gut during bacterial fermentation of dietary fiber and is attributed numerous beneficial effects on the host metabolism. We aimed to develop a method for the assessment of functional capacity of gut microbiota butyrate synthesis based on the qPCR quantification of bacterial gene coding butyryl-CoA:acetate CoA-transferase, the key enzyme of butyrate synthesis. In silico, we identified bacteria possessing but gene among human gut microbiota by searching but coding sequences in available databases. We designed and validated six sets of degenerate primers covering all selected bacteria, based on their phylogenetic nearness and sequence similarity, and developed a method for gene abundance normalization in human fecal DNA. We determined but gene abundance in fecal DNA of subjects with opposing dietary patterns and metabolic phenotypes—lean vegans (VG) and healthy obese omnivores (OB) with known fecal microbiota and metabolome composition. We found higher but gene copy number in VG compared with OB, in line with higher fecal butyrate content in VG group. We further found a positive correlation between the relative abundance of target bacterial genera identified by next-generation sequencing and groups of but gene-containing bacteria determined by specific primers. In conclusion, this approach represents a simple and feasible tool for estimation of microbial functional capacity. View Full-Text

Keywords: gut microbiota; butyrate; functional capacity

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Daskova, N.; Heczkova, M.; Modos, I.; Videnska, P.; Splichalova, P.; Pelantova, H.; Kuzma, M.; Gojda, J.; Cahova, M. Determination of Butyrate Synthesis Capacity in Gut Microbiota: Quantification of but Gene Abundance by qPCR in Fecal Samples. Biomolecules 2021, 11, 1303. https://doi.org/10.3390/biom11091303

AMA Style

Daskova N, Heczkova M, Modos I, Videnska P, Splichalova P, Pelantova H, Kuzma M, Gojda J, Cahova M. Determination of Butyrate Synthesis Capacity in Gut Microbiota: Quantification of but Gene Abundance by qPCR in Fecal Samples. Biomolecules. 2021; 11(9):1303. https://doi.org/10.3390/biom11091303

Chicago/Turabian Style

Daskova, Nikola, Marie Heczkova, Istvan Modos, Petra Videnska, Petra Splichalova, Helena Pelantova, Marek Kuzma, Jan Gojda, and Monika Cahova. 2021. "Determination of Butyrate Synthesis Capacity in Gut Microbiota: Quantification of but Gene Abundance by qPCR in Fecal Samples" Biomolecules 11, no. 9: 1303. https://doi.org/10.3390/biom11091303

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Determination of Butyrate Synthesis Capacity in Gut Microbiota: Quantification of but Gene Abundance by qPCR in Fecal Samples

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