scholarly journals Noninvasive imaging and quantification of bile salt hydrolase activity: From bacteria to humans

2021 ◽  
Vol 7 (6) ◽  
pp. eaaz9857
Author(s):  
Pavlo V. Khodakivskyi ◽  
Christian L. Lauber ◽  
Aleksey Yevtodiyenko ◽  
Arkadiy A. Bazhin ◽  
Stephen Bruce ◽  
...  
Keyword(s):  
Bile Salt ◽  
Noninvasive Imaging ◽  
Clinical Status ◽  
Cost Effective ◽  
Bile Salt Hydrolase ◽  
Inflammatory Bowel ◽  
Bile Salt Hydrolase Activity ◽  
Bsh Activity ◽  
Entire Gastrointestinal Tract

The microbiome-produced enzyme bile salt hydrolase (BSH) plays a central role in human health, but its function remains unclear due to the lack of suitable methods for measuring its activity. Here, we have developed a novel optical tool based on ultrasensitive bioluminescent imaging and demonstrated that this assay can be used for quick and cost-effective quantification of BSH activity across a broad range of biological settings including pure enzymes and bacteria, intact fecal slurries, and noninvasive imaging in live animals, as well as for the assessment of BSH activity in the entire gastrointestinal tract of mice and humans. Using this assay, we showed that certain types of prebiotics are capable of increasing BSH activity of the gut microbiota in vivo and successfully demonstrated potential application of this assay as a noninvasive diagnostic test to predict the clinical status of inflammatory bowel disease (IBD) patients.

scholarly journals Functional Analysis of Four Bile Salt Hydrolase and Penicillin Acylase Family Members in Lactobacillus plantarum WCFS1

2008 ◽  
Vol 74 (15) ◽  
pp. 4719-4726 ◽  
Author(s):  
Jolanda M. Lambert ◽  
Roger S. Bongers ◽  
Willem M. de Vos ◽  
Michiel Kleerebezem
Keyword(s):  
Bile Salt ◽  
Lactobacillus Plantarum ◽  
Bile Salts ◽  
Natural Habitat ◽  
Penicillin Acylase ◽  
Exponential Growth Phase ◽  
Bile Salt Hydrolase ◽  
Intestinal Microbes ◽  
Bsh Activity

ABSTRACT Bile salts play an important role in the digestion of lipids in vertebrates and are synthesized and conjugated to either glycine or taurine in the liver. Following secretion of bile salts into the small intestine, intestinal microbes are capable of deconjugating the glycine or taurine from the bile salts, using an enzyme called bile salt hydrolase (Bsh). Intestinal lactobacilli are regarded as major contributors to bile salt hydrolysis in vivo. Since the bile salt-hydrolyzing strain Lactobacillus plantarum WCFS1 was predicted to carry four bsh genes (bsh1, bsh2, bsh3, and bsh4), the functionality of these bsh genes was explored using Lactococcus lactis heterologous overexpression and multiple bsh deletion strains. Thus, Bsh1 was shown to be responsible for the majority of Bsh activity in L. plantarum WCFS1. In addition, bsh1 of L. plantarum WCFS1 was shown to be involved in conferring tolerance to specific bile salts (i.e., glycocholic acid). Northern blot analysis established that bsh1, bsh2, bsh3, and bsh4 are all expressed in L. plantarum WCFS1 during the exponential growth phase. Following biodiversity analysis, bsh1 appeared to be the only bsh homologue that was variable among L. plantarum strains; furthermore, the presence of bsh1 correlated with the presence of Bsh activity, suggesting that Bsh1 is commonly responsible for Bsh activity in L. plantarum strains. The fact that bsh2, bsh3, and bsh4 genes appeared to be conserved among L. plantarum strains suggests an important role of these genes in the physiology and lifestyle of the species L. plantarum. Analysis of these additional bsh-like genes in L. plantarum WCFS1 suggests that they might encode penicillin acylase rather than Bsh activity, indicating their implication in the conversion of substrates other than bile acids in the natural habitat.

scholarly journals Cholesterol-Lowering Potentials of Lactic Acid Bacteria Based on Bile-Salt Hydrolase Activity and Effect of Potent Strains on Cholesterol MetabolismIn VitroandIn Vivo

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Cheng-Chih Tsai ◽  
Pei-Pei Lin ◽  
You-Miin Hsieh ◽  
Zi-yi Zhang ◽  
Hui-Ching Wu ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Bile Acid ◽  
Lactic Acid Bacteria ◽  
Bile Salt ◽  
Hepg2 Cells ◽  
Bile Salt Hydrolase ◽  
Apo B ◽  
Probiotic Strains ◽  
Bile Salt Hydrolase Activity ◽  
Bsh Activity

This study collected different probiotic isolates from animal and plant sources to evaluate the bile-salt hydrolase activity of probioticsin vitro. The deconjugation potential of bile acid was determined using high-performance liquid chromatography. HepG2 cells were cultured with probiotic strains with high BSH activity. The triglyceride (TG) and apolipoprotein B (apo B) secretion by HepG2 cells were evaluated. Our results show that the BSH activity and bile-acid deconjugation abilities ofPediococcus acidilacticiNBHK002,Bifidobacterium adolescentisNBHK006,Lactobacillus rhamnosusNBHK007, andLactobacillus acidophilusNBHK008 were higher than those of the other probiotic strains. The cholesterol concentration in cholesterol micelles was reduced within 24 h. NBHK007 reduced the TG secretion by 100% after 48 h of incubation. NBHK002, NBHK006, and NBHK007 could reduce apo B secretion by 33%, 38%, and 39%, respectively, after 24 h of incubation. The product PROBIO S-23 produced a greater decrease in the total concentration of cholesterol, low-density lipoprotein, TG, and thiobarbituric acid reactive substance in the serum or livers of hamsters with hypercholesterolemia compared with that of hamsters fed with a high-fat and high-cholesterol diet. These results show that the three probiotic strains of lactic acid bacteria are better candidates for reducing the risk of cardiovascular disease.

scholarly journals Development of a Covalent Inhibitor of Gut Bacterial Bile Salt Hydrolases

2019 ◽  
Author(s):  
Arijit A. Adhikari ◽  
Tom C. Seegar ◽  
Scott B. Ficarro ◽  
Megan D. McCurry ◽  
Deepti Ramachandran ◽  
...  
Keyword(s):  
Bile Acid ◽  
Bile Salt ◽  
Bile Acids ◽  
Unmet Need ◽  
Cysteine Residue ◽  
Gut Bacteria ◽  
Bile Salt Hydrolase ◽  
Secondary Bile Acid ◽  
Bsh Activity

AbstractBile salt hydrolase (BSH) enzymes are widely expressed by human gut bacteria and catalyze the gateway reaction leading to secondary bile acid formation. Bile acids regulate key metabolic and immune processes by binding to host receptors. There is an unmet need for a potent tool to inhibit BSHs across all gut bacteria in order to study the effects of bile acids on host physiology. Here, we report the development of a covalent pan-inhibitor of gut bacterial BSH. From a rationally designed candidate library, we identified a lead compound bearing an alpha-fluoromethyl ketone warhead that modifies BSH at the catalytic cysteine residue. Strikingly, this inhibitor abolished BSH activity in conventional mouse feces. Mice gavaged with a single dose of this compound displayed decreased BSH activity and decreased deconjugated bile acid levels in feces. Our studies demonstrate the potential of a covalent BSH inhibitor to modulate bile acid composition in vivo.

scholarly journals A gut-restricted lithocholic acid analog as an inhibitor of gut bacterial bile salt hydrolases

2021 ◽  
Author(s):  
Arijit A. Adhikari ◽  
Deepti Ramachandran ◽  
Snehal N. Chaudhari ◽  
Chelsea E. Powell ◽  
Megan D. McCurry ◽  
...  
Keyword(s):  
Bile Acid ◽  
Bile Salt ◽  
Bile Acids ◽  
Mammalian Cells ◽  
Lithocholic Acid ◽  
Bile Salt Hydrolase ◽  
Host Physiology ◽  
Secondary Bile Acids ◽  
Bsh Activity

AbstractBile acids play crucial roles in host physiology by acting as both detergents that aid in digestion and as signaling molecules that bind to host receptors. Gut bacterial bile salt hydrolase (BSH) enzymes perform the gateway reaction leading to the conversion of host-produced primary bile acids into bacterially modified secondary bile acids. Small molecule probes that target BSHs will help elucidate the causal roles of these metabolites in host physiology. We previously reported the development of a covalent BSH inhibitor with low gut permeability. Here, we build on our previous findings and describe the development of a second-generation gut-restricted BSH inhibitor with enhanced potency, reduced off-target effects, and durable in vivo efficacy. SAR studies focused on the bile acid core identified a compound, AAA-10, containing a C3-sulfonated lithocholic acid scaffold and an alpha-fluoromethyl ketone warhead as a potent pan-BSH inhibitor. This compound inhibits BSH activity in conventional mouse fecal slurries, bacterial cultures, and purified BSH proteins and displays reduced toxicity against mammalian cells compared to first generation compounds. Oral administration of AAA-10 to wild-type mice for 5 days resulted in a decrease in the abundance of the secondary bile acids deoxycholic acid (DCA) and lithocholic acid (LCA) in the mouse GI tract with low systemic exposure of AAA-10, demonstrating that AAA-10 is an effective tool for inhibiting BSH activity and modulating bile acid pool composition in vivo.

Bile Salt Hydrolase Activity of Enterococci Isolated from Food: Screening and Quantitative Determination

2001 ◽  
Vol 64 (5) ◽  
pp. 725-729 ◽  
Author(s):  
CHARLES M. A. P. FRANZ ◽  
INGRID SPECHT ◽  
PETRA HABERER ◽  
WILHELM H. HOLZAPFEL
Keyword(s):  
Bile Salt ◽  
Bile Salt Hydrolase ◽  
Screening Assay ◽  
Screening Experiments ◽  
Enterococcus Casseliflavus ◽  
Enterococcus Gallinarum ◽  
Activity Groups ◽  
De Man ◽  
Bile Salt Hydrolase Activity ◽  
Bsh Activity

One hundred seventeen enterococcal strains isolated from food (47 Enterococcus faecium, 48 Enterococcus faecalis, 16 Enterococcus durans, 2 Enterococcus gallinarum, 3 Enterococcus casseliflavus, and 1 Enterococcus malodoratus) were screened for bile salt hydrolase (BSH) activity on de Man, Rogosa, and Sharpe agar medium containing taurocholic acid and calcium chloride. The highest incidence of BSH-active strains was observed for E. faecalis (81%) followed by E. faecium (50%) and E. durans (44%). Isolates were grouped into four putative activity groups (no, low, medium, and high activity) based on the size of precipitation zones observed in the screening experiment. Our results showed that assumptions on BSH activity based on the size of bile precipitation zones in screening experiments did not correlate with actual activity as quantified by high-pressure liquid chromatography, but the screening assay is useful for assessing the presence or absence of BSH activity.

scholarly journals Quantitative Determination of Bile Salt Hydrolase Activity in Bacteria Isolated from the Small Intestine of Chickens

2002 ◽  
Vol 68 (12) ◽  
pp. 6425-6428 ◽  
Author(s):  
Ane Knarreborg ◽  
Ricarda M. Engberg ◽  
Søren K. Jensen ◽  
Bent B. Jensen
Keyword(s):  
Small Intestine ◽  
Bile Salt ◽  
High Performance ◽  
Bile Salt Hydrolase ◽  
Protein Determination ◽  
Chromatography Analysis ◽  
Fermentation Profile ◽  
Bile Salt Hydrolase Activity ◽  
Bsh Activity

ABSTRACT A quantitative assay based on high-performance liquid chromatography analysis of bile salts and bacterial protein determination was established for investigating bile salt hydrolase (BSH) activity in bacteria isolated from the small intestine of chickens. Bacteria were isolated using various media and were subsequently grouped according to cell morphology, fermentation profile, and 16S ribosomal DNA sequence. Representative isolates from each bacterial group were assayed for BSH activity. The isolates differed in BSH activity with respect to the state of growth and preculturing with and without taurochenodeoxycholate. The highest levels of BSH activity were found with Enterococcus faecium and Clostridium perfringens.

scholarly journals In Vitro Bile Salt Hydrolase (BSH) Activity Screening of Different Probiotic Microorganisms

Foods ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 674
Author(s):  
Jimmy G. Hernández-Gómez ◽  
Argelia López-Bonilla ◽  
Gabriela Trejo-Tapia ◽  
Sandra V. Ávila-Reyes ◽  
Antonio R. Jiménez-Aparicio ◽  
...  
Keyword(s):  
Bile Salt ◽  
High Performance ◽  
Probiotic Strain ◽  
Saccharomyces Boulardii ◽  
Bile Salt Hydrolase ◽  
Sodium Glycocholate ◽  
Probiotic Yeast ◽  
Probiotic Strains ◽  
Bsh Activity

Bile salt hydrolase (BSH) activity in probiotic strains is usually correlated with the ability to lower serum cholesterol levels in hypercholesterolemic patients. The objective of this study was the evaluation of BSH in five probiotic strains of lactic acid bacteria (LAB) and a probiotic yeast. The activity was assessed using a qualitative direct plate test and a quantitative high-performance thin- layer chromatography assay. The six strains differed in their BSH substrate preference and activity. Lactobacillus plantarum DGIA1, a potentially probiotic strain isolated from a double cream cheese from Chiapas, Mexico, showed excellent deconjugation activities in the four tested bile acids (69, 100, 81, and 92% for sodium glycocholate, glycodeoxycholate, taurocholate, and taurodeoxycholate, respectively). In the case of the commercial probiotic yeast Saccharomyces boulardii, the deconjugation activities were good against sodium glycodeoxycholate, taurocholate, and taurodeoxycholate (100, 57, and 63%, respectively). These last two results are part of the novelty of the work. A weak deconjugative activity (5%) was observed in the case of sodium glycocholate. This is the first time that the BSH activity has been detected in this yeast.

scholarly journals Compact Miniaturized Bioluminescence Sensor Based on Continuous Air-Segmented Flow for Real-Time Monitoring: Application to Bile Salt Hydrolase (BSH) Activity and ATP Detection in Biological Fluids

Chemosensors ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 122
Author(s):  
Aldo Roda ◽  
Pierpaolo Greco ◽  
Patrizia Simoni ◽  
Valentina Marassi ◽  
Giada Moroni ◽  
...  
Keyword(s):  
Bile Salt ◽  
Light Emission ◽  
Ex Vivo ◽  
Biological Fluids ◽  
Low Cost ◽  
Bile Salt Hydrolase ◽  
Segmented Flow ◽  
Active Substrate ◽  
Flat Spiral ◽  
Bsh Activity

A simple and versatile continuous air-segmented flow sensor using immobilized luciferase was designed as a general miniaturized platform based on sensitive biochemiluminescence detection. The device uses miniaturized microperistaltic pumps to deliver flows and compact sensitive light imaging detectors based on BI-CMOS (smartphone camera) or CCD technology. The low-cost components and power supply make it suitable as out-lab device at point of need to monitor kinetic-related processes or ex vivo dynamic events. A nylon6 flat spiral carrying immobilized luciferase was placed in front of the detector in lensless mode using a fiber optic tapered faceplate. ATP was measured in samples collected by microdialysis from rat brain with detecting levels as low as 0.4 fmoles. The same immobilized luciferase was also used for the evaluation of bile salt hydrolase (BSH) activity in intestinal microbiota. An aminoluciferin was conjugatated with chenodeoxycholic acid forming the amide derivative aLuc-CDCA. The hydrolysis of the aLuc-CDCA probe by BSH releases free uncaged aminoluciferin which is the active substrate for luciferase leading to light emission. This method can detect as low as 0.5 mM of aLuc-CDCA, so it can be used on real faecal human samples to study BSH activity and its modulation by diseases and drugs.

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