Milk samples were skimmed twice by centrifugation at 8,500g for 30 min and then subjected to in vitro digestion (13) . After protein concentration was determined by the Bradford method (37), simulated gastric digestion was started with adjusting the pH to 4.0 (mimicking the stomach pH in infants) with 1 mol/l HCl. Porcine pepsin (Sigma-Aldrich, St Louis, MO; 2% in 1 mmol/l HCl) was added to the sample in a 1:12.5 ratio (pepsin:protein). Samples were placed in an incubating shaker (New Brunswick Scientific, Edison, NJ) at 140 rpm at 37 °C for 15 min. Then, after the pH of the samples was adjusted to 7.0 with 0.1 mol/l NaHCO 3 , simulated intestinal digestion was performed. Pancreatin (Sigma-Aldrich; 0.4% in 0.1 mol/l NaHCO 3 ) was added to the samples in a 1:62.5 ratio (Pancreatin:protein), and the samples were placed in the incubator shaker at 140 rpm at 37 °C for 5 min. After the incubation, the enzymes were inactivated in a water bath at 85 °C for 3 min.
Pancreatin
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Poland, China, Italy, France, Spain, Switzerland, Canada, Australia, India, Ireland, Jersey, Belgium, Sao Tome and Principe, Denmark
Pancreatin is a digestive enzyme complex derived from the pancreas of mammals. It contains a mixture of digestive enzymes, including amylase, lipase, and protease, which play a role in the breakdown of carbohydrates, fats, and proteins, respectively.
Automatically generated - may contain errors
Lab products found in correlation
Pepsin, by Merck Group (270 mentions)
Bile salt, by Merck Group (78 mentions)
α amylase, by Merck Group (64 mentions)
Hydrochloric acid (hcl), by Merck Group (37 mentions)
Trypsin, by Merck Group (31 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (31 mentions)
Collagenase type 2, by Worthington (29 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (28 mentions)
Sodium chloride (nacl), by Merck Group (26 mentions)
Sodium hydroxide (naoh), by Merck Group (24 mentions)
Gallic acid, by Merck Group (22 mentions)
Lipase, by Merck Group (20 mentions)
Bovine serum albumin (bsa), by Merck Group (19 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (18 mentions)
Methanol, by Merck Group (17 mentions)
Bile extract, by Merck Group (17 mentions)
Mucin, by Merck Group (16 mentions)
Sodium bicarbonate, by Merck Group (16 mentions)
Formic acid, by Merck Group (15 mentions)
Penicillin, by Thermo Fisher Scientific (15 mentions)
520 protocols using pancreatin
1
In Vitro Infant Milk Digestion
2
Standardized Flavonoids and Phenolic Acids Analysis
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Standard (purity > 98%) flavonoids and phenolic acids including quinic acid, gallic acid, protocatechuic acid, epicatechin, 5-caffeoylquinic acid, catechin, p-cumaric acid, apigenin-7-O-glucoside, ferulic acid, naringin, rutin, quercetin 3 galattoside, hesperidin, rosamarinic acid, kaempferol 3 glucoside, ellagic acid, diosmin, genistein, isorhamnetin 3 rutinoside, myricetin, daidzein, quercetin, naringenin, gallic acid, luteolin, and apigenin were provided from Sigma-Aldrich (Milan, Italy). For the antioxidant experiments, 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS), 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox), gallic acid, potassium persulphate, 2,3,5-triphenyltetrazolio chloride (TPTZ), 1,1-diphenyl-2-picrylhydrazyl (DPPH), anhydrous ferric chloride, sodium acetate, and hydrochloric acid were provided from Sigma-Aldrich. Standards and enzymes used to simulate GI process were: pepsin, bile salt, α-amylase, pancreatin, bacterial protease from Streptomyces griseus (Pronase E), Viscozyme L and pancreatin were provided from Sigma-Aldrich. Moreover, Megazyme® phytic acid assay kit, ammonium molybdate, potassium persulfate, calcium chloride, sodium carbonate, hydrochloric acid, acetate buffer, ferric chloride (FeCl3) and sodium chloride were provided from Sigma-Aldrich.
3
Stability of Cabbage and Eudragit-Coated PDVs
4
Chia Seed Protein Purification
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The chia seeds used for the study were collected from a local market in Mysuru, Karnataka, India. The proteases, Pepsin and Pancreatin were procured from MERCK (SIGMA-ALDRICH). HiTrap chelating HP immobilized metal affinity chromatography columns were procured from GE Healthcare Biosciences. All the chemicals used were of analytical grade and solvents were of HPLC grade.
5
Flaxseed Oil Cake Characterization and Evaluation
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Flaxseed oil cake (FOC) was purchased from ACS Sp. z o.o. (Bydgoszcz, Poland). The proximate composition of FOC (based on supplier information) was: solids—80.50%, including: proteins—41.97%; carbohydrates—27.99%; fiber—6.29%; fat—6.11%; ash—4.50%. Lacticaseibacillus rhamnosus GG (ATCC53103), was procured from ATCC (Manassas, VA, USA). Buffered peptone water, microbiological agar, MRS agar and broth were purchased from Oxoid (Basingstoke, UK). Potassium chloride, sodium chloride, potassium thiocyanate, disodium hydrogen phosphate, monosodium dihydrogen orthophosphate, calcium chloride, sodium hydrogen carbonate, hydrochloric acid, ammonium chloride, sodium peroxide, urea, α-amylase, uric acid, mucin from porcine stomach (type II), glucose, glucuronic acid, glucosamine hydrochloride, bovine serum albumin (BSA), pepsin, pancreatin, oxgall, Triton X-100, cholesterol, ethanol, ninhydrin, glacial acetic acid, cadmium chloride and hexadecane were purchased from Merck Chemical (Saint Louis, MI, USA). All reagents were of analytical grade.
6
Evaluating Probiotic Survival Under Simulated Gut Conditions
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According to Huang and Adams, (2004) (link),
in vitro simulated gastrointestinal tract (GIT)
conditions were prepared. Similarly, gastric juice was prepared by mixing 1
mL of sodium chloride (0.7%) (Sigma-Aldrich®,
India) and pepsin (3 mg) (Sigma-Aldrich®, India), with a
final pH in between 2 or 3. Whereas, simulated intestinal juice was prepared
by mixing pancreatin (1 mg) (Merck®), 1 mL of sodium
chloride (0.7%) (Sigma-Aldrich®, India),
1.5% of bile salts (Sigma-Aldrich®, India) with
final pH 8.
E. durans F3 was grown (0.6–0.7 OD at 600 nm) into
fresh MRS broth by inoculating the log phase culture, followed by
centrifugation of cell culture (8,000 rpm, 10 min, 4°C). Washing of
cell pellet was then performed by resuspending the cells in sterile
phosphate buffered saline (PBS) (pH 7.0). Resuspended cells (0.5 mL) were
added into 2.0 mL of simulated gastric or intestinal juices with incubation
at 37°C for 6 h. Survival cell count were calculated at 0 (initial
time) and 2, 4, and 6 h by spreading the 0.1 mL of cell suspension on MRS
agar plates with incubation at 37°C. Following formula was used to
calculate the survival percentage of E. durans F3 under
simulated gastric juices:
in vitro simulated gastrointestinal tract (GIT)
conditions were prepared. Similarly, gastric juice was prepared by mixing 1
mL of sodium chloride (0.7%) (Sigma-Aldrich®,
India) and pepsin (3 mg) (Sigma-Aldrich®, India), with a
final pH in between 2 or 3. Whereas, simulated intestinal juice was prepared
by mixing pancreatin (1 mg) (Merck®), 1 mL of sodium
chloride (0.7%) (Sigma-Aldrich®, India),
1.5% of bile salts (Sigma-Aldrich®, India) with
final pH 8.
E. durans F3 was grown (0.6–0.7 OD at 600 nm) into
fresh MRS broth by inoculating the log phase culture, followed by
centrifugation of cell culture (8,000 rpm, 10 min, 4°C). Washing of
cell pellet was then performed by resuspending the cells in sterile
phosphate buffered saline (PBS) (pH 7.0). Resuspended cells (0.5 mL) were
added into 2.0 mL of simulated gastric or intestinal juices with incubation
at 37°C for 6 h. Survival cell count were calculated at 0 (initial
time) and 2, 4, and 6 h by spreading the 0.1 mL of cell suspension on MRS
agar plates with incubation at 37°C. Following formula was used to
calculate the survival percentage of E. durans F3 under
simulated gastric juices:
7
Rapeseed Digestion Protocol
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A batch of commercially available winter 00-rapeseed (Brassica napus), harvested in the southwest of France in 2013, was used. All chemicals used were of analytical grade. Pepsin (2,000 FIP U/g) was obtained from Merck (Darmstadt, Germany), whilst pancreatin (grade IV from porcine pancreas), trypsin (type IX-S, 13,000–20,000 BAEE units/mg protein), chymotrypsin (type II, >40 units/mg protein), and peptidase from porcine intestinal mucosa (50–100 units/g solid) were obtained from Sigma-Aldrich (St Louis, MO, USA).
8
Enzymatic Hydrolysis of Angiotensin-Converting Enzyme
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Low molecular weight (LMW) chitosan, sodium tripolyphosphate, angiotensin I-converting enzyme from rabbit lung and N-hippuryl-histidyl-leucine were purchased from Sigma Aldrich (St. Louis, MO, USA). Bromelain, Pepsin, Trypsin, Pancreatin, Tween 80, mono and dibasic potassium salts, sodium chloride, and sodium hydroxide were obtained from Merck (Darmstadt & Schuchardt, Germany). All other chemicals used were of analytical grade and purchased from Fisher scientific (Loughborough, UK), R & M (Essex, UK), Merck (Darmstadt, Germany) and Sigma Aldrich (St. Louis, MO, USA).
9
Comprehensive Phytochemical Analysis of Plant Extracts
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Hydrochloric acid (HCl), sodium phosphate monobasic, sodium phosphate dibasic, catechol, Folin–Ciocalteu’s phenol reagent, gallic acid, methanol, ethanol, sodium carbonate, formic acid, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), 2,4,6-tripyridyl-striazine, Iron(III) chloride hexahydrate, sodium acetate anhydrous, acetic acid, sodium hydroxide, potassium hydroxide (KOH), guar gum, methanol, 2,2-diphenyl-1-picrylhydrazyl (DPPH), phytochemical standards (quinic acid, chlorogenic acid, caffeic acid, rutin, 4-coumaric acid, ferulic acid), and the enzymes (pepsin, invertase, amyloglucosidase, and pancreatin) were purchased from Merck Ltd. (formerly Sigma Aldrich Ltd., Wicklow, Ireland).
10
Cell Line Culture Conditions
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The human CML cell line K562 was purchased from the China Center for Type Culture Collection (Wuhan, China) and cultured in RPMI (Hyclone, Logan, UT, USA) supplemented with 10% FBS (NQBB, Adelaide, Australia). The T293 cell line was cultured in DMEM (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS in an incubator at 37°C with 5% CO2. Cells were passaged with pancreatin (Merck, St Louis, MO, USA) when approximately 70%–90% confluence was reached. Imatinib was kindly supplied by Novartis International AG (Basel, Switzerland).
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