Quantitative assay to detect bacterial glycan-degrading enzyme activities in mouse and human fecal samples

Alex Steimle; Erica T Grant; Mahesh S Desai

doi:10.1016/j.xpro.2021.100326

Quantitative assay to detect bacterial glycan-degrading enzyme activities in mouse and human fecal samples

STAR Protoc. 2021 Feb 17;2(1):100326. doi: 10.1016/j.xpro.2021.100326. eCollection 2021 Mar 19.

Authors

Alex Steimle¹, Erica T Grant^{1

2}, Mahesh S Desai^{1

3}

Affiliations

¹ Department of Infection and Immunity, Luxembourg Institute of Health, Esch-sur-Alzette 4354, Luxembourg.
² Faculty of Science, Technology and Medicine, University of Luxembourg, Esch-sur-Alzette 4365, Luxembourg.
³ Odense Research Center for Anaphylaxis, Department of Dermatology and Allergy Center, Odense University Hospital, University of Southern Denmark, 5000 Odense, Denmark.

Abstract

The gut microbiome expresses a multitude of enzymes degrading polysaccharides in dietary plant fibers and in host-secreted mucus. The quantitative detection of these glycan-degrading enzymes in fecal samples is important to elucidate the functional activity of the microbiome in health and disease. We describe a protocol for detection of glycan-degrading enzyme activity in mouse and human fecal samples, namely sulfatase and four carbohydrate-active enzymes. Assessing their activity can inform treatment strategies for diseases linked to the gut microbiome. For complete details on the use and execution of this protocol, please refer to Desai et al. (2016).

Keywords: Metabolism; Protein expression and purification.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Bacteria / enzymology*
Bacterial Proteins / metabolism*
Feces* / enzymology
Feces* / microbiology
Glycoside Hydrolases / metabolism*
Humans
Mice
Microbiota*

Substances

Bacterial Proteins
Glycoside Hydrolases
glycanase