RD43 rice (Oryza sativa L.) and Hom Mali 105 rice (Oryza sativa L.) were obtained from the Rice Department, Bangkok, Thailand. Pepsin from porcine gastric mucosa powder (250 U/mg), α-amylase Type VI-B from porcine pancreas (15.8 U/mg), pancreatin from porcine pancreas (4×U.S. Pharmacopeia (USP) specifications), pure amylose from potatoes, phosphatidylcholine, oleic acid, taurocholic acid, glycodeoxycholic acid, taurodeoxycholic acid, taurocholic acid, and porcine cholesterol esterase (35 U/mg) were purchased from Sigma-Aldrich, Inc. (St. Louis, MO, United States). Amyloglucosidase (3,260 U/mL) was purchased from Megazyme International Ireland Ltd. (Bray, Ireland). Glucose oxidase kit (Glucose liquicolor) and Cholesterol test kits (Cholesterol liquicolor) were purchased from Human (Human diagnostics, Wiesbaden, Germany). The total bile acid kit was purchased from BIOBASE (Jinan, Shandong, China).
Taurocholic acid
Manufactured by Merck Group
Sourced in United States, United Kingdom, Switzerland, Germany, Sao Tome and Principe
Taurocholic acid is a bile acid that functions as a surfactant, facilitating the digestion and absorption of fats in the small intestine. It is a key component in the bile produced by the liver and plays a role in the emulsification and solubilization of lipids.
Automatically generated - may contain errors
Lab products found in correlation
Taurodeoxycholic acid, by Merck Group (38 mentions)
Taurochenodeoxycholic acid, by Merck Group (31 mentions)
Glycocholic acid, by Merck Group (28 mentions)
Glycodeoxycholic acid, by Merck Group (27 mentions)
Cholic acid, by Merck Group (26 mentions)
Chenodeoxycholic acid, by Merck Group (26 mentions)
Deoxycholic acid, by Merck Group (24 mentions)
Glycochenodeoxycholic acid, by Merck Group (24 mentions)
Lithocholic acid, by Merck Group (19 mentions)
Ursodeoxycholic acid, by Merck Group (18 mentions)
Taurolithocholic acid, by Merck Group (14 mentions)
Tauroursodeoxycholic acid, by Merck Group (14 mentions)
Glycoursodeoxycholic acid, by Merck Group (8 mentions)
Oleic acid, by Merck Group (7 mentions)
Phosphatidylcholine, by Merck Group (6 mentions)
Porcine pancreatic lipase, by Merck Group (5 mentions)
Porcine cholesterol esterase, by Merck Group (5 mentions)
Bovine serum albumin (bsa), by Merck Group (5 mentions)
Orlistat, by Merck Group (4 mentions)
Hyodeoxycholic acid, by Merck Group (4 mentions)
91 protocols using taurocholic acid
1
In Vitro Digestion of Rice Cultivars
2
Comprehensive Bile Acid Standards Analysis
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Glycholithocholic acid (GLCA) standards were purchased from Toronto Research Chemicals; Cholic acid (CA), lithocholic acid (LCA), deoxycholic acid (DCA), ursodeoxycholic acid (UDCA), chenodeoxycholic acid (CDCA), taurocholic acid (TCA), glychocholic acid (GCA), taurolithocholic acid (TLCA), taurodeoxycholic acid (TDCA), tauroursodeoxycholic acid (TUDCA), taurochenodeoxycholic acid (TCDCA), glychodeoxycholic acid (GDCA), glychochenodeoxycholic acid (GCDCA), glychoursodeoxycholic acid (GUDCA) standards, and isotope-labeled internal standard deuterocholic acid (d4-CA), deuterodeoxycholic acid (d4-DCA), deuteroglychocholic acid (d4-GCA), deuterated ursodeoxycholic acid (d4-UDCA), and deuterated lithocholic acid (d4-LCA) were purchased from Sigma Aldrich; Chromatographic pure acetonitrile and chromatographic pure methanol were purchased from Merck, Germany; Chromatographic grade formic acid was purchased from Shanghai Aladdin.
3
Culture and Maintenance of Esophageal Cell Lines
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Human esophageal epithelial cells, STR, were cultured as previously described. In brief, Keratinocyte-SFM medium was supplemented with 1 ng/mL Epidermal Growth Factor, 0.05 mg/mL Bovine Pituitary Extract, and 1% penicillin streptomycin antibiotics (Gibco™, for Life Technologies, Inc., Carlsbad, CA, USA). Bile/acid-tolerant STR cells were established by repeated exposure to a 100 μM of cocktail of bile salts (20 μM of each, deoxycholic acid (MP Biomedicals, Santa Ana, CA, USA), glycocholic acid, taurocholic acid, sodium glycodeoxycholate and sodium glycochenodeoxycholate (all Sigma, St. Louis, MO, USA) as reported) [34 (link)].The non-neoplastic Barrett’s epithelial cell lines CPA, CPB, BAR-T and BAR-10T cells were isolated from non-dysplastic metaplasia (kind gift from Dr. Rhonda Souza, Baylor Scott White) [35 (link)], and were cultured in Epithelial Cell Medium-2 supplemented with 5% epithelial cell growth supplement-2 (EpiCGS-2) and 5% penicillin/streptomycin antibiotics (ScienceCell™ Research Laboratories, Carlsbad, CA, USA). The esophageal adenocarcinoma cell lines, FLO1, OE33 and OE19 (kind gift of Dr. El-Rifai, University of Miami), were grown in RPMI with 10% FBS and 1% penicillin/streptomycin antibiotics (ThermoFisher Scientific, Waltham, MA, USA). All cells were incubated at 37 °C with 5% CO2.
4
Measurement of Digestive Enzyme Activity
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Gallic acid, rat intestinal acetone powder, porcine pancreatic α-amylase, 4-methylumbelliferone, glucose oxidase kits and 3,5-dinitrosalicylic acid p-nitrophenylbutylrate (p-NPB), oleic acid, phosphatidylcholine, glycodeoxycholic acid, taurodeoxycholic acid, taurocholic acid, porcine cholesterol esterase, porcine pancreatic lipase were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Cholesterol test kits were purchased from HUMAN GmbH Co. (Wiesbaden, Germany). Total bile acid kit was purchased from Bio-Quant Co. (San Diego, CA, USA). All other chemical reagents used in this study were of analytical grade.
5
Cecal Bile Acid Quantification
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Cecal contents (5 controls and 8 from L. paracasei treated mice) from trial 3 were used to quantify primary and secondary bile acids. Bile acids analysis was performed at Bioaster (Lyon, France). Samples were prepared from 50 to 60 mg of −80 °C frozen cecal content using aqueous methanol extraction process. Bile acids extracts were analysed by LC-MS/MS on a triple quadrupole Thermo Quantum Ultra (SN: TQU00665) combined to a Dionex Ultimate 3000 HPLC system (SN: 8074045 & 8087183). Samples were separated on a C18 column (2,7 μM, 150 × 2,1 mm) from Ascentis Express using a methanol/water gradient containing 5 mM ammonium acetate and 0,012% formic acid. All bile acid standards: LithoCholic acid, ChenoDeoxyCholic acid, DeoxyCholic acid, UrsoDeoxyCholic acid, Cholic acid, GlycoDeoxyCholic acid, GlycoursoDeoxyCholic acid, GlyChenoDeoxyCholic acid, Glychocolic acid, TauroLithoCholic acid, TauroDeoxyCholic acid, TauroCholic acid, were bought from Sigma Aldrich. Results of bile salt quantification were expressed in peak area, corrected for sample weight.
6
Bile Acid Preparation and Characterization
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BAM8-22 was purchased from Tocris (Bristol, UK) and dissolved in 1× phosphate buffered saline (PBS). Cholic acid (CA), tauro-Cholic acid (TCA), glycol-Cholic acid (GCA), chenodeoxyCholic acid (CDCA), tauro-chenodeoxyCholic acid (TCDCA), glycol-chenodeoxyCholic acid (GCDCA), deoxyCholic acid (DCA), tauro-deoxyCholic acid (TDCA), lithoCholic acid (LCA), and tauro-lithoCholic acid (TLCA) were purchased from Sigma Aldrich (St. Louis, Missouri, USA). Protease inhibitor cocktail (PIC) was purchased from Bio Vision (Milpitas, CA 95035 USA).
7
Assessing Bifidobacterium Bile Tolerance
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To determine Bifidobacterium survival in bile, isolates were first grown in RCM and then subcultured using a 1:50 dilution into MRS ± 0.3% unfractionated bovine bile salt (Sigma-Aldrich), as described by [57 (link)]. After 48 h of stationary growth in an anaerobic chamber at 37 °C OD600 nm using the Benchmark Plus microplate spectrophotometer (Bio-Rad) for both conditions. For the MRS plate the mean blank OD600 nmvalue was 0.1585 and for the anaerobic plate it was 0.1825. Data shown are mean values from three experimental repeats.To assess bile salt hydrolyase activity, overnight cultures were spotted (3 mL) onto MRS plates supplemented with l -cysteine and 0.5% w/v of either taurocholic acid, taurodeoxycholic acid, and sodium glycodeoxycholate bile salt (Sigma-Aldrich). Bile salt precipitation was assessed after a maximum of 96 h of anaerobic incubation at 37 °C. For both assays, uninoculated MRS media were used as a control.
8
Evaluating Bifidobacterium Bile Tolerance
9
Purification and Mutagenesis of Bile Salt Hydrolase
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Major bacterial strains and plasmids used in this study and their sources are listed in Table S1 . Briefly, the E. coli recombinant strain JL885 containing pBSH expression vector (Table S1 ), constructed in our previous study19 (link), was used for purification of wild-type lsBSH in this study. The pBSH plasmid bearing original bsh gene from L. salivarius NRRL B-30514 was used as parent vector for site-directed mutagenesis. All compounds were purchased from Sigma-Aldrich (St. Louis, MO, USA), which include ampicillin, glycocholic acid (GCA), glycodeoxycholic acid (GDCA), glycochenodeoxycholic acid (GCDCA), taurocholic acid (TCA), taurodeoxycholic acid (TDCA), taurochenodeoxycholic acid (TCDCA).
10
Acid and Bile Tolerance of Probiotic Strain
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For acid tolerance, CNCM I-4884 suspension was adjusted to 1.0 (OD 600 nm) with phosphate buffered saline (PBS) at pH 2.0 using 1 N HCl. After 30, 60, 90 and 120 min of incubation at 37 °C, suspensions were serially diluted in PBS and plated onto MRS agar. Viable cells were counted after 48 h incubation at 37 °C. For bile salts tolerance, MRS broth was supplemented with a mix of taurocholic acid, taurochenodeoxycholic acid, glycocholic acid and glycochenodeoxycholic acid (Sigma, Saint Louis, MO, USA) at 0.05 mg/mL, 0.1 mg/mL, 0.5 mg/mL, 1 mg/mL and 2 mg/mL each. Bacterial growth was monitored at 37 °C every 30 min for 24 h by OD at 600 nm using Infinite M200 Pro spectrophotometer (TECAN, Lyon, France). The experiments were performed in three biological replicates.
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