Bile salts b8756

Soybean oil (SBO) and pomegranate oil (PGO) were provided by CJ-Cheil-Jedang Co. (Seoul, Korea) and Kerfoot Inc. (North Yorkshire, UK), respectively. Immobilized lipase from Rhizomucor miehei (Lipozyme^® RMIM) was obtained from Novozymes Korea (Seoul, Korea). Pancreatin (P3293) and type II lipase (L3126) from porcine pancreases, bile salts (B8756), porcine bile extract (B8631), and bovine serum albumin (BSA, A7906) were obtained from Sigma-Aldrich Korea (Yongin, Korea). Triundecanoin was purchased from Nu-Chek-Prep (Waterville, ME, USA). Sodium chloride (NaCl), sodium bicarbonate (NaHCO₃), monopotassium phosphate (KH₂PO₄), potassium chloride (KCl), magnesium chloride (MgCl₂), calcium chloride (CaCl₂), hydrogen chloride (HCl) and sodium hydroxide solution (0.05 N, NaOH) were purchased from Daejung Chemicals & Metals Co. (Siheung, Korea). Urea was purchased from Junsei Chemical Co. (Tokyo, Japan). The solvents used for HPLC and gas chromatography (GC) analysis were purchased from Fisher Scientific Korea (Seoul, Korea).

In order to determine MIC of erythromycin, tetracycline and streptomycin, M90T wild-type strain and its derivatives M90T ΔacrAB, M90T ΔacrA, M90T ΔacrB pSfacrB_D408A, and M90T ΔacrD, were inoculated into LB and grown at 37 °C by shaking for 16 h. Cultures were then diluted to OD₆₀₀ 0.02 in LB and 100 µL aliquots were transferred to 96-well plate, each well containing 100 µL of 2-fold serial dilutions of the compounds to be tested (erythromycin 0.001 mg/mL to 2 mg/mL, tetracycline 0.0078 μg/mL to 16 μg/mL, and streptomycin 0.0625 μg/mL to 128 μg/mL). After 16 h incubation at 37 °C, the lowest concentration of antibiotic inhibiting bacterial growth was estimated, as previously described [40 (link)]. At least three biological replicates were performed. Bile salts (B8756, Sigma-Aldrich, St. Louis, MO, USA), consisting of an approximate 1:1 mixture of cholate and deoxycholate, were used at 0.25% (w/v).

Fermented noni juice (FNJ) with ~7% solids was obtained from a health store (Auckland, New Zealand). Carriers including maltodextrin (MD: 10–13 DE) and gum acacia (GA) were kindly given by Ingredion Ltd. (Auckland, New Zealand). HPLC (High-Performance Liquid Chromatography)-grade acetonitrile was bought from JT Baker (Phillipsburg, NJ, USA), formic acid was purchased from Romil Ltd. (Cambridge, UK), Folin–Ciocalteus’s phenol reagent and acetic acid were purchased from Merck KGaA (Darmstadt, Germany) and 2,2-azino-biz-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) diammonium salt was from Abcam (Cambridge, UK). Reagents for in vitro digestion including sodium acetate, 8x USP pancreatin from porcine pancreas (P7545), pepsin from porcine gastric mucosa (P7000) and bile salts (B8756) were purchased from Sigma-Aldrich (Auckland, New Zealand). Ammonium carbonate, magnesium chloride hexahydrate, potassium persulfate and sodium carbonate were from ECP Ltd. (Auckland, New Zealand), calcium chloride dehydrate (AR grade) was from Thermo Fisher Scientific (Auckland, New Zealand) and sodium hydrogen carbonate (AR grade) was obtained from VWR (Auckland, New Zealand). All water used was of Milli-Q grade.