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Title: A continuous fluorescence assay for simple quantification of bile salt hydrolase activity in the gut microbiome

Abstract

The microbiota of the mammalian gut plays a dynamic role in controlling host physiology. The effect of gut microbiota activity on host health is particularly evident in the case of bile homeostasis. Bile is produced by the host and is modified by the gut microbiota, which impacts the net hydrophobicity of the total bile acid pool, and also modulates host signaling pathways. A key mechanism by which the microbiota modify bile is through de-conjugation of bile salts through bile salt hydrolase (BSH) enzymatic activity, which is postulated to be a prerequisite for all further microbial metabolism. BSH activity in the gut is largely considered to be beneficial, and genes encoding BSHs are found in the genomes of many taxa found in over-the-counter probiotics. Despite the therapeutic relevance of this enzyme, there is no reliable and simple assay for continuous monitoring of BSH activity, and there are no non-destructive means of characterizing its activity in whole cell or microbial community samples. Herein, we describe a continuous fluorescence assay that can be used for characterization of BSH activity with purified protein, cell lysates, whole cells, and in human gut microbiome samples. The method is a “turn-on” reporter strategy, which employs synthetic substratesmore » that yield a fluorescent product upon BSH-dependent turnover. This assay is used to show the first in vivo characterization of BSH activity. Furthermore, we demonstrate the first example of continuous, non-destructive quantification of BSH activity in human gut microbiome samples.« less

Authors:
 [1];  [1]; ORCiD logo [1];  [1]
  1. Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1501853
Report Number(s):
PNNL-SA-133128
Journal ID: ISSN 2045-2322
Grant/Contract Number:  
AC05-76RL01830
Resource Type:
Accepted Manuscript
Journal Name:
Scientific Reports
Additional Journal Information:
Journal Volume: 9; Journal Issue: 1; Journal ID: ISSN 2045-2322
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Brandvold, Kristoffer R., Weaver, Jacqueline M., Whidbey, Christopher, and Wright, Aaron T. A continuous fluorescence assay for simple quantification of bile salt hydrolase activity in the gut microbiome. United States: N. p., 2019. Web. doi:10.1038/s41598-018-37656-7.
Brandvold, Kristoffer R., Weaver, Jacqueline M., Whidbey, Christopher, & Wright, Aaron T. A continuous fluorescence assay for simple quantification of bile salt hydrolase activity in the gut microbiome. United States. doi:10.1038/s41598-018-37656-7.
Brandvold, Kristoffer R., Weaver, Jacqueline M., Whidbey, Christopher, and Wright, Aaron T. Mon . "A continuous fluorescence assay for simple quantification of bile salt hydrolase activity in the gut microbiome". United States. doi:10.1038/s41598-018-37656-7. https://www.osti.gov/servlets/purl/1501853.
@article{osti_1501853,
title = {A continuous fluorescence assay for simple quantification of bile salt hydrolase activity in the gut microbiome},
author = {Brandvold, Kristoffer R. and Weaver, Jacqueline M. and Whidbey, Christopher and Wright, Aaron T.},
abstractNote = {The microbiota of the mammalian gut plays a dynamic role in controlling host physiology. The effect of gut microbiota activity on host health is particularly evident in the case of bile homeostasis. Bile is produced by the host and is modified by the gut microbiota, which impacts the net hydrophobicity of the total bile acid pool, and also modulates host signaling pathways. A key mechanism by which the microbiota modify bile is through de-conjugation of bile salts through bile salt hydrolase (BSH) enzymatic activity, which is postulated to be a prerequisite for all further microbial metabolism. BSH activity in the gut is largely considered to be beneficial, and genes encoding BSHs are found in the genomes of many taxa found in over-the-counter probiotics. Despite the therapeutic relevance of this enzyme, there is no reliable and simple assay for continuous monitoring of BSH activity, and there are no non-destructive means of characterizing its activity in whole cell or microbial community samples. Herein, we describe a continuous fluorescence assay that can be used for characterization of BSH activity with purified protein, cell lysates, whole cells, and in human gut microbiome samples. The method is a “turn-on” reporter strategy, which employs synthetic substrates that yield a fluorescent product upon BSH-dependent turnover. This assay is used to show the first in vivo characterization of BSH activity. Furthermore, we demonstrate the first example of continuous, non-destructive quantification of BSH activity in human gut microbiome samples.},
doi = {10.1038/s41598-018-37656-7},
journal = {Scientific Reports},
number = 1,
volume = 9,
place = {United States},
year = {2019},
month = {2}
}

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