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 »
- Publication Date:
- Research Org.:
- Pacific Northwest National Laboratory (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. https://doi.org/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. https://doi.org/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 = {Mon Feb 04 00:00:00 EST 2019},
month = {Mon Feb 04 00:00:00 EST 2019}
}
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Cited by: 13 works
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Figures / Tables:
Figure 1: Design of an assay to quantify the activity of an enzyme that is a key regulator of bile acid homeostasis. (A) Bile salt hydrolase activity of the gut microbiota modulates the properties of bile salts, which are produced by the liver. BSH facilitates hydrolysis of the amide bondmore »
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Works referenced in this record:
Bile acids and the gut microbiome
journal, January 2014
- Ridlon, Jason M.; Kang, Dae J.; Hylemon, Phillip B.
- Current Opinion in Gastroenterology, Vol. 30, Issue 3
New fluorescent bile acids: Synthesis, chemical characterization, and disastereoselective uptake by Caco-2 cells of 3-deoxy 3-NBD-amino deoxycholic and ursodeoxycholic acid
journal, March 2012
- Májer, Ferenc; Salomon, Johanna J.; Sharma, Ruchika
- Bioorganic & Medicinal Chemistry, Vol. 20, Issue 5
Human nutrition, the gut microbiome and the immune system
journal, June 2011
- Kau, Andrew L.; Ahern, Philip P.; Griffin, Nicholas W.
- Nature, Vol. 474, Issue 7351
A combinatorial approach for determining protease specificities: application to interleukin-1β converting enzyme (ICE)
journal, January 1997
- Rano, Thomas A.; Timkey, Tracy; Peterson, Erin P.
- Chemistry & Biology, Vol. 4, Issue 2
The gut microbiome in health and in disease
journal, January 2015
- Shreiner, Andrew B.; Kao, John Y.; Young, Vincent B.
- Current Opinion in Gastroenterology, Vol. 31, Issue 1
Deconjugation of bile salts by Lactobacillus acidophilus isolates
journal, January 2003
- Ahn, Young Tae; Kim, Geun Bae; Lim, Kwang Sei
- International Dairy Journal, Vol. 13, Issue 4
Regulation of host weight gain and lipid metabolism by bacterial bile acid modification in the gut
journal, May 2014
- Joyce, S. A.; MacSharry, J.; Casey, P. G.
- Proceedings of the National Academy of Sciences, Vol. 111, Issue 20
Seasonal cycling in the gut microbiome of the Hadza hunter-gatherers of Tanzania
journal, August 2017
- Smits, Samuel A.; Leach, Jeff; Sonnenburg, Erica D.
- Science, Vol. 357, Issue 6353
Quantitative Determination of Bile Salt Hydrolase Activity in Bacteria Isolated from the Small Intestine of Chickens
journal, December 2002
- Knarreborg, A.; Engberg, R. M.; Jensen, S. K.
- Applied and Environmental Microbiology, Vol. 68, Issue 12
Antibiotic growth promoters enhance animal production by targeting intestinal bile salt hydrolase and its producers
journal, January 2014
- Lin, Jun
- Frontiers in Microbiology, Vol. 5
Vitamin D Receptor As an Intestinal Bile Acid Sensor
journal, May 2002
- Makishima, M.
- Science, Vol. 296, Issue 5571
Biological deoxycholic acid–coumarin conjugates: photo-switchable structures and self-assembly morphology
journal, May 2016
- Sun, Jinzhi; Xiang, Shuaishuai; Xu, Xiang
- Tetrahedron Letters, Vol. 57, Issue 19
Key discoveries in bile acid chemistry and biology and their clinical applications: history of the last eight decades
journal, May 2014
- Hofmann, Alan F.; Hagey, Lee R.
- Journal of Lipid Research, Vol. 55, Issue 8
Bile salt hydrolase and cholesterol removal effect by Bifidobacterium bifidum NRRL 1976
journal, November 2005
- Parvez, S.; Kim, Hong-Yeoul; Lee, Han-Chang
- World Journal of Microbiology and Biotechnology, Vol. 22, Issue 5
New fluorescent bile acids: Synthesis, chemical characterization, and disastereoselective uptake by Caco-2 cells of 3-deoxy 3-NBD-amino deoxycholic and ursodeoxycholic acid
journal, March 2012
- Májer, Ferenc; Salomon, Johanna J.; Sharma, Ruchika
- Bioorganic & Medicinal Chemistry, Vol. 20, Issue 5
Discovery of Bile Salt Hydrolase Inhibitors Using an Efficient High-Throughput Screening System
journal, January 2014
- Smith, Katie; Zeng, Ximin; Lin, Jun
- PLoS ONE, Vol. 9, Issue 1
Structural and Functional Analysis of a Conjugated Bile Salt Hydrolase from Bifidobacterium longum Reveals an Evolutionary Relationship with Penicillin V Acylase
journal, August 2006
- Kumar, R. Suresh; Brannigan, James A.; Prabhune, Asmita A.
- Journal of Biological Chemistry, Vol. 281, Issue 43
Host-Gut Microbiota Metabolic Interactions
journal, June 2012
- Nicholson, J. K.; Holmes, E.; Kinross, J.
- Science, Vol. 336, Issue 6086
A substrate combinatorial array for caspases
journal, June 1999
- Lee, Dennis; Adams, Jerry L.; Brandt, Martin
- Bioorganic & Medicinal Chemistry Letters, Vol. 9, Issue 12
Bile Salt Hydrolase Activity in Probiotics
journal, March 2006
- Begley, M.; Hill, C.; Gahan, C. G. M.
- Applied and Environmental Microbiology, Vol. 72, Issue 3
Bile Salt Hydrolase Activity in Probiotics
journal, March 2006
- Begley, M.; Hill, C.; Gahan, C. G. M.
- Applied and Environmental Microbiology, Vol. 72, Issue 3
Characterization and purification of bile salt hydrolase from Lactobacillus sp. strain 100-100.
journal, August 1990
- Lundeen, S. G.; Savage, D. C.
- Journal of Bacteriology, Vol. 172, Issue 8
Characterization and purification of bile salt hydrolase from Lactobacillus sp. strain 100-100.
journal, August 1990
- Lundeen, S. G.; Savage, D. C.
- Journal of Bacteriology, Vol. 172, Issue 8
Development of a differential medium for bile salt hydrolase-active Lactobacillus spp.
journal, January 1989
- Dashkevicz, M. P.; Feighner, S. D.
- Applied and Environmental Microbiology, Vol. 55, Issue 1
Seasonal cycling in the gut microbiome of the Hadza hunter-gatherers of Tanzania
journal, August 2017
- Smits, Samuel A.; Leach, Jeff; Sonnenburg, Erica D.
- Science, Vol. 357, Issue 6353
Fluorescent bile acid derivatives: Relationship between chemical structure and hepatic and intestinal transport in the rat
journal, November 1997
- Holzinger, F.; Schteingart, C. D.; Ton-Nu, H.
- Hepatology, Vol. 26, Issue 5
Regulation of host weight gain and lipid metabolism by bacterial bile acid modification in the gut
journal, May 2014
- Joyce, S. A.; MacSharry, J.; Casey, P. G.
- Proceedings of the National Academy of Sciences, Vol. 111, Issue 20
A core gut microbiome in obese and lean twins
journal, November 2008
- Turnbaugh, Peter J.; Hamady, Micah; Yatsunenko, Tanya
- Nature, Vol. 457, Issue 7228
Intestinal Crosstalk between Bile Acids and Microbiota and Its Impact on Host Metabolism
journal, July 2016
- Wahlström, Annika; Sayin, Sama I.; Marschall, Hanns-Ulrich
- Cell Metabolism, Vol. 24, Issue 1
Bile acid biosynthesis
journal, May 1992
- Russell, David W.; Setchell, Kenneth D. R.
- Biochemistry, Vol. 31, Issue 20
Bile salt biotransformations by human intestinal bacteria
journal, November 2005
- Ridlon, Jason M.; Kang, Dae-Joong; Hylemon, Phillip B.
- Journal of Lipid Research, Vol. 47, Issue 2
Bile Salt Hydrolase of Bifidobacterium longum---Biochemical and Genetic Characterization
journal, June 2000
- Tanaka, H.; Hashiba, H.; Kok, J.
- Applied and Environmental Microbiology, Vol. 66, Issue 6
The gut microbiota, bacterial metabolites and colorectal cancer
journal, September 2014
- Louis, Petra; Hold, Georgina L.; Flint, Harry J.
- Nature Reviews Microbiology, Vol. 12, Issue 10
Confocal Imaging with a Fluorescent Bile Acid Analogue Closely Mimicking Hepatic Taurocholate Disposition
journal, June 2014
- De Bruyn, Tom; Sempels, Wouter; Snoeys, Jan
- Journal of Pharmaceutical Sciences, Vol. 103, Issue 6
Confocal Imaging with a Fluorescent Bile Acid Analogue Closely Mimicking Hepatic Taurocholate Disposition
journal, June 2014
- De Bruyn, Tom; Sempels, Wouter; Snoeys, Jan
- Journal of Pharmaceutical Sciences, Vol. 103, Issue 6
An improved colorimetric determination of amino acids with the use of ninhydrin
journal, January 1966
- Lee, Ya Pin; Takahashi, Tunekazu
- Analytical Biochemistry, Vol. 14, Issue 1
Sensitive assays for trypsin, elastase, and chymotrypsin using new fluorogenic substrates
journal, March 1977
- Zimmerman, M.; Ashe, B.; Yurewicz, E. C.
- Analytical Biochemistry, Vol. 78, Issue 1
A G Protein-coupled Receptor Responsive to Bile Acids
journal, January 2003
- Kawamata, Yuji; Fujii, Ryo; Hosoya, Masaki
- Journal of Biological Chemistry, Vol. 278, Issue 11
Sensitive assays for trypsin, elastase, and chymotrypsin using new fluorogenic substrates
journal, March 1977
- Zimmerman, M.; Ashe, B.; Yurewicz, E. C.
- Analytical Biochemistry, Vol. 78, Issue 1
Endogenous Bile Acids Are Ligands for the Nuclear Receptor FXR/BAR
journal, May 1999
- Wang, Haibo; Chen, Jasmine; Hollister, Kevin
- Molecular Cell, Vol. 3, Issue 5
Structural comparison of Ntn-hydrolases
journal, January 2000
- Oinonen, Carita; Rouvinen, Juha
- Protein Science, Vol. 9, Issue 12
All 4 Bile Salt Hydrolase Proteins Are Responsible for the Hydrolysis Activity in Lactobacillus plantarum ST-III
journal, November 2011
- Ren, Jing; Sun, Kejie; Wu, Zhengjun
- Journal of Food Science, Vol. 76, Issue 9
Discovery of Bile Salt Hydrolase Inhibitors Using an Efficient High-Throughput Screening System
journal, January 2014
- Smith, Katie; Zeng, Ximin; Lin, Jun
- PLoS ONE, Vol. 9, Issue 1
Bile acid biosynthesis: Pathways and regulation
journal, October 1974
- Mosbach, Erwin H.; Salen, Gerald
- The American Journal of Digestive Diseases, Vol. 19, Issue 10
Bile salt hydrolase and cholesterol removal effect by Bifidobacterium bifidum NRRL 1976
journal, November 2005
- Parvez, S.; Kim, Hong-Yeoul; Lee, Han-Chang
- World Journal of Microbiology and Biotechnology, Vol. 22, Issue 5
Bile Salt Hydrolase of Bifidobacterium longum---Biochemical and Genetic Characterization
journal, June 2000
- Tanaka, H.; Hashiba, H.; Kok, J.
- Applied and Environmental Microbiology, Vol. 66, Issue 6
All 4 Bile Salt Hydrolase Proteins Are Responsible for the Hydrolysis Activity in Lactobacillus plantarum ST-III
journal, November 2011
- Ren, Jing; Sun, Kejie; Wu, Zhengjun
- Journal of Food Science, Vol. 76, Issue 9
Bile acids: regulation of synthesis
journal, April 2009
- Chiang, John Y. L.
- Journal of Lipid Research, Vol. 50, Issue 10
Fluorescent bile acid derivatives: Relationship between chemical structure and hepatic and intestinal transport in the rat
journal, November 1997
- Holzinger, F.; Schteingart, C. D.; Ton-Nu, H.
- Hepatology, Vol. 26, Issue 5
Bile acid biosynthesis
journal, May 1992
- Russell, David W.; Setchell, Kenneth D. R.
- Biochemistry, Vol. 31, Issue 20
Identification of membrane-type receptor for bile acids (M-BAR)
journal, November 2002
- Maruyama, Takaharu; Miyamoto, Yasuhisa; Nakamura, Takao
- Biochemical and Biophysical Research Communications, Vol. 298, Issue 5
Quantitative Determination of Bile Salt Hydrolase Activity in Bacteria Isolated from the Small Intestine of Chickens
journal, December 2002
- Knarreborg, A.; Engberg, R. M.; Jensen, S. K.
- Applied and Environmental Microbiology, Vol. 68, Issue 12
Biological deoxycholic acid–coumarin conjugates: photo-switchable structures and self-assembly morphology
journal, May 2016
- Sun, Jinzhi; Xiang, Shuaishuai; Xu, Xiang
- Tetrahedron Letters, Vol. 57, Issue 19
Bile acids and the gut microbiome
journal, January 2014
- Ridlon, Jason M.; Kang, Dae J.; Hylemon, Phillip B.
- Current Opinion in Gastroenterology, Vol. 30, Issue 3
Side chain conjugation prevents bacterial 7-dehydroxylation of bile acids.
journal, July 1990
- Batta, A. K.; Salen, G.; Arora, R.
- Journal of Biological Chemistry, Vol. 265, Issue 19
Works referencing / citing this record:
Chemoproteomic Profiling of Gut Microbiota-Associated Bile Salt Hydrolase Activity
journal, April 2019
- Parasar, Bibudha; Zhou, Hao; Xiao, Xieyue
- ACS Central Science
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