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Casein hydrolysate
Casein hydrolysate
Casein hydrolysate is a protein-rich product derived from the enzymatic digestion of casein, the primary protein found in milk.
It contains a complex mixture of amino acids, peptides, and other bioactive compounds that have a variety of applications in the food, pharmaceutical, and nutraceutical industries.
Casein hydrolysate is commonly used as a dietary supplement, a source of protein in sports nutrition products, and as an ingredient in specialized medical foods.
Its unique compositoin and properties make it a valuable tool for researchers exploring its potential health benefits, such as its ability to support muscle recovery, bone health, and immune function.
Optimizing the production and characterization of casein hydrolysate is an important area of study, and PubCompare.ai can help researchers easily locate and compare relevant protocols from the scientific literature, preprints, and patents to identify the best methods and products for their casein hydrolysate research.
It contains a complex mixture of amino acids, peptides, and other bioactive compounds that have a variety of applications in the food, pharmaceutical, and nutraceutical industries.
Casein hydrolysate is commonly used as a dietary supplement, a source of protein in sports nutrition products, and as an ingredient in specialized medical foods.
Its unique compositoin and properties make it a valuable tool for researchers exploring its potential health benefits, such as its ability to support muscle recovery, bone health, and immune function.
Optimizing the production and characterization of casein hydrolysate is an important area of study, and PubCompare.ai can help researchers easily locate and compare relevant protocols from the scientific literature, preprints, and patents to identify the best methods and products for their casein hydrolysate research.
Most cited protocols related to «Casein hydrolysate»
Amino Acids
Amino Acid Sequence
Anti-Inflammatory Agents
Antihypertensive Agents
Biological Processes
Biopharmaceuticals
Caseins
Debility
Domestic Sheep
DPP4 protein, human
Gene Products, Protein
Goat
Homo sapiens
Immunomodulation
Microbicides
Milk, Cow's
Milk Proteins
Opioids
Parent
Peptides
Proteins
Psychological Inhibition
Staphylococcal Protein A
Whey
For electroporation experiments, protoplasts were isolated from five-day old Nicotiana tabacum BY-2 suspension cells. Suspension cultures were grown at 23°C with shaking (130 rpm) in a medium containing Murashige and Skoog salts [21 (link)] supplemented with 1 mg/L thiamine-HCl, 370 mg/L KH2PO4, 30 g/L sucrose, and 2 mg/L2, 4-dichlorophenoxyacetic acid, pH 5.7. Cells were sub-cultured once per week by adding 2.5–3 ml of inoculum to 50 ml of fresh medium in 250 ml Erlenmeyer flasks. The 50 ml of suspension culture was centrifuged at 250 × g at room temperature for 5 min in a Sorvall GLC-2 centrifuge, and the pellet (15 ml packed cell volume) was re-suspended in 50 ml of protoplast isolation solution containing 7.4 g/L CaCl2 ·2H2O, 1 g/L NaOAc, and 45 g/L mannitol supplemented with 1.2% cellulose R10 (Onazuka) and 0.6% Macerozyme (Duchefa), pH 5.7. Approximately 15 ml of suspension culture was transferred into three 20 × 100 mm sterile Petri dishes and incubated in the dark with a gentle shaking (40 rpm) at room temperature for 4 hours. The protoplasts were washed twice with protoplast isolation solution and the pellet was re-suspended in 50 ml of floating solution (99 mg/L myo-inositol, 2.88 g/L L-proline, 100 mg/L enzymatic casein hydrolysate, 102.6 g/L sucrose, 97.6 mg/L MES buffer, 4.3 g/L MS salts, 1 mg/L thiamine-HCl, 370 mg/L KH2PO4, pH 5.7). Protoplasts (floating on the top of the solution) were transferred to a new tube, washed twice, and suspended in 50 ml of electroporation solution (10 mM NaCl, 4 mM CaCl2·2H2O, 120 mM KCl, 10 mM HEPES, 0.6 M mannitol, pH 7.2). Cells were incubated at 42°C for 5 min and kept in ice for 10 min before electroporation. Aliquots of protoplasts containing approximately 3 × 106 cells/ml were used for electroporation. ~20 μg of each plasmid DNA were added to 300 μl of protoplasts in a tube and placed on ice. The electroporation was conducted using a BioRad Gene Pulser apparatus at 0.16 kV, with the Pulse Controller set to infinity and the capacitance extender set to 960 μFD. After 10 min incubation on ice, the protoplasts were transferred into 10 ml of BY-2 culture medium supplemented with 0.4 M mannitol and incubated overnight.
For direct DNA uptake experiments, 20 ml BY-2 suspension cells were transferred to a sterile conical centrifuge tube and centrifuged at 3000 rpm for 10 min. The pelleted cells were suspended in 10–20 ml protoplast digestion enzyme solution (1.2% Cellulase Onozuka RS [Duchefa] and 0.6% Macerozyme R-10 [Duchefa] in 10 mM CaCl2·2H2O, 12 mM NaOAc, 11% mannitol, pH 5.7) and incubated in the dark with shaking (40 rpm) for 3–4 hr at room temperature. The protoplasts were filtered through 40 μm nylon mesh and centrifuged in a 50 ml conical tube at 250 × g for 5 min. The protoplasts were collected and suspended in 10 ml protoplast floating solution (per liter: 99 mg myo-inositol, 2.88 g L-proline, 100 mg enzymatic casein hydrolysate, 102.6 g sucrose, 97.6 mg MES buffer, 4.3 g MS salts, 1 mg Vitamin B1, 370 mg KH2PO4, pH 5.7) and centrifuged at 250 × g for 10 min. Protoplasts floating in this solution were removed and 10 ml W5 solution (154 mM NaCl, 125 mM CaCl2, 5 mM KCl, 2 mM MES, pH 5.7) was added. The solution was centrifuged at 250 × g for 5 min, and the protoplasts pelleted. Protoplast concentration was adjusted to 1 × 106/ml and the solution incubated on ice for 30 min. The protoplasts were again centrifuged at 250 × g for 5 min and suspended at a density of 1 × 106 cells/ml in MMg solution (0.6 M mannitol, 15 mM MgCl2, 4 mM MES, pH 5.7). DNA (10 μg in 10 μl) was added to 100 μl protoplasts, followed by addition of 110 μl PEG solution (per ml: 0.4 g PEG 4000 [Fluca], 0.6 ml 1 M mannitol, 100 μl 1 M CaCl2), and the protoplasts incubated at room temperature for 30 min. After addition of 1 ml W5 solution, the protoplast suspension was centrifuged at 250 × g for 5 min. The protoplast pellet was suspended in 1 ml incubation solution (per liter: MS salts and vitamins, 2 mg/L 2,4-D, 3% sucrose, 0.4 ~ 0.6 M mannitol) and incubated in the dark for 16 hr at 26°C.
Particle bombardment of onion epidermal peel layers was carried out using a Biolistic Particle Delivery System (Bio-Rad) PDS-1000. Samples (whole onion from which the dry outer layer was removed) were sterilized in ~300 ml 2% NaOCl and 2–3 drops Tween-20 for 15 min. The tissue was washed with sterilize H2O at least five times. The upper epidermal layer of the onion was removed, cut into 2 × 2 cm squares, and placed on a plate containing 1/2 MS medium. 5 μg of each plasmid DNA was used in all experiments. Gold particles size was 1.6 μm (INBIO GOLD). 0.15–0.2 mg particles/per shot were used with a chamber vacuum of 27 in Hg. Particles were accelerated with a pressure of 1100 psi. The distance between the projectile source and the samples was 6 cm.
For direct DNA uptake experiments, 20 ml BY-2 suspension cells were transferred to a sterile conical centrifuge tube and centrifuged at 3000 rpm for 10 min. The pelleted cells were suspended in 10–20 ml protoplast digestion enzyme solution (1.2% Cellulase Onozuka RS [Duchefa] and 0.6% Macerozyme R-10 [Duchefa] in 10 mM CaCl2·2H2O, 12 mM NaOAc, 11% mannitol, pH 5.7) and incubated in the dark with shaking (40 rpm) for 3–4 hr at room temperature. The protoplasts were filtered through 40 μm nylon mesh and centrifuged in a 50 ml conical tube at 250 × g for 5 min. The protoplasts were collected and suspended in 10 ml protoplast floating solution (per liter: 99 mg myo-inositol, 2.88 g L-proline, 100 mg enzymatic casein hydrolysate, 102.6 g sucrose, 97.6 mg MES buffer, 4.3 g MS salts, 1 mg Vitamin B1, 370 mg KH2PO4, pH 5.7) and centrifuged at 250 × g for 10 min. Protoplasts floating in this solution were removed and 10 ml W5 solution (154 mM NaCl, 125 mM CaCl2, 5 mM KCl, 2 mM MES, pH 5.7) was added. The solution was centrifuged at 250 × g for 5 min, and the protoplasts pelleted. Protoplast concentration was adjusted to 1 × 106/ml and the solution incubated on ice for 30 min. The protoplasts were again centrifuged at 250 × g for 5 min and suspended at a density of 1 × 106 cells/ml in MMg solution (0.6 M mannitol, 15 mM MgCl2, 4 mM MES, pH 5.7). DNA (10 μg in 10 μl) was added to 100 μl protoplasts, followed by addition of 110 μl PEG solution (per ml: 0.4 g PEG 4000 [Fluca], 0.6 ml 1 M mannitol, 100 μl 1 M CaCl2), and the protoplasts incubated at room temperature for 30 min. After addition of 1 ml W5 solution, the protoplast suspension was centrifuged at 250 × g for 5 min. The protoplast pellet was suspended in 1 ml incubation solution (per liter: MS salts and vitamins, 2 mg/L 2,4-D, 3% sucrose, 0.4 ~ 0.6 M mannitol) and incubated in the dark for 16 hr at 26°C.
Particle bombardment of onion epidermal peel layers was carried out using a Biolistic Particle Delivery System (Bio-Rad) PDS-1000. Samples (whole onion from which the dry outer layer was removed) were sterilized in ~300 ml 2% NaOCl and 2–3 drops Tween-20 for 15 min. The tissue was washed with sterilize H2O at least five times. The upper epidermal layer of the onion was removed, cut into 2 × 2 cm squares, and placed on a plate containing 1/2 MS medium. 5 μg of each plasmid DNA was used in all experiments. Gold particles size was 1.6 μm (INBIO GOLD). 0.15–0.2 mg particles/per shot were used with a chamber vacuum of 27 in Hg. Particles were accelerated with a pressure of 1100 psi. The distance between the projectile source and the samples was 6 cm.
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All media recipes were made at pH 7 and autoclaved with the appropriate starting ingredients. NZY media for 1 L was made with 5 g yeast extract, 10 g casein hydrolysate, 5 g NaCL, and 1 g MgSO4. NZYB consisted of NZY plus phosphate buffer. TB media for 1 L was made containing 12 g yeast extract, 24 g casein hydrolysate, 4 mL glycerol, and phosphate buffer. FL media for 1 L was comprised of 10 g yeast extract, 20 g casein hydrolysate, 5 g NaCL, 1 g MgSO4, 20 mL glycerol, and phosphate buffer. Supplementation with glycerol and phosphate buffer occurred at the time of induction.
All transformations were carried out using E. coli strain C41 Overexpress (Lucigen), an optimal strain for terpene synthase expression (Prisic and Peters 2007 (link)). Transformations consisting of three or more plasmids required 0.5 µg plasmid DNA in 50 µL aliquots of chemically competent E. coli C41 cells. The resulting recombinant strains were grown under selective conditions using the appropriate antibiotics at concentrations of 25 µg/mL for carbenicillin, 20 µg/mL for chloramphenicol, 15 µg/mL tetracycline, and 15 µg/mL spectinomycin. For all concentrations listed, negative controls lacking resistance to one marker confirmed the ability of the lower antibiotic concentrations to fully inhibit growth. Expression growths were initiated from several colonies inoculated together into liquid media. For all shake flasks cultures, initial growth to log phase was carried out at 37 °C (A600 ∼0.6), with the temperature then dropped to 16 °C, whereupon the pH was adjusted to 7.0 and phosphate buffer and glycerol, where described, were added. Shaking was held at 200 rpm, and the cultures induced 1 h after dropping the temperature to 16 °C by the addition of IPTG to 1 mM. Baffled flasks were tested under the same conditions as regular Erlenmeyer flasks.
During growth, cultures were intermittently monitored for pH, cell density, and where applicable, product output (as indicated in the various figures). Cell density was monitored by absorbance of aliquots (one in ten dilutions) at 600 nm in a Varian spectrophotometer. Culture pH was monitored throughout growth and adjusted, if necessary, to ensure that the culture pH remained in the 6.5 to 7.5 range, although the cultures rarely required adjustments after the shift to 16 °C. Supplementation with pyruvate or mevalonolactone was administered via pulse feeding with 1 M solutions at regular intervals of 12 h for the first 36 h, up to the desired maximum concentration. Investigation of the role of cell density on product formation (Fig.3 ) was conducted in 48 h parallel aliquots for each density, from the time of IPTG induction, whereby the cell densities were controlled by moving the culture in aliquots from the initial incubation temperature of 37 to 16 °C, where it was held for the remainder of the time course. The values recorded for the cell density are the average of the two aliquots; the percent variance between the duplicate cell densities was smaller than the percent variance between the determined abietadiene yield, therefore error in abietadiene yield is displayed. Note that the data reported in Tables 1 and 2 , as well as Figs. 2 , 3 , 4 , 5 , 7 , and 8 , were derived from parallel fermentation runs (i.e., all the data reported in each of these came from cultures grown together in the same incubator, each in duplicate).
Bioreactor growths were conducted in a New Brunswick BioFlo110 fermentor, set up according the manufacturer's instructions. Under our settings, the cultures were stirred at 300 rpm, their temperature maintained at 20 °C, pH held at 7.2 (using 5 M KOH and 5 M HCL reservoirs connected to the A and B pumps), chemically resistant tubing was used, and air flow was maintained at 4 lpm using triple micron filtered air. Pulse feeding supplements were introduced through the feeding septa, and cell density and product output monitored, as well as pH verified, by intermittent sample removal for subsequent measurement.
All transformations were carried out using E. coli strain C41 Overexpress (Lucigen), an optimal strain for terpene synthase expression (Prisic and Peters 2007 (link)). Transformations consisting of three or more plasmids required 0.5 µg plasmid DNA in 50 µL aliquots of chemically competent E. coli C41 cells. The resulting recombinant strains were grown under selective conditions using the appropriate antibiotics at concentrations of 25 µg/mL for carbenicillin, 20 µg/mL for chloramphenicol, 15 µg/mL tetracycline, and 15 µg/mL spectinomycin. For all concentrations listed, negative controls lacking resistance to one marker confirmed the ability of the lower antibiotic concentrations to fully inhibit growth. Expression growths were initiated from several colonies inoculated together into liquid media. For all shake flasks cultures, initial growth to log phase was carried out at 37 °C (A600 ∼0.6), with the temperature then dropped to 16 °C, whereupon the pH was adjusted to 7.0 and phosphate buffer and glycerol, where described, were added. Shaking was held at 200 rpm, and the cultures induced 1 h after dropping the temperature to 16 °C by the addition of IPTG to 1 mM. Baffled flasks were tested under the same conditions as regular Erlenmeyer flasks.
During growth, cultures were intermittently monitored for pH, cell density, and where applicable, product output (as indicated in the various figures). Cell density was monitored by absorbance of aliquots (one in ten dilutions) at 600 nm in a Varian spectrophotometer. Culture pH was monitored throughout growth and adjusted, if necessary, to ensure that the culture pH remained in the 6.5 to 7.5 range, although the cultures rarely required adjustments after the shift to 16 °C. Supplementation with pyruvate or mevalonolactone was administered via pulse feeding with 1 M solutions at regular intervals of 12 h for the first 36 h, up to the desired maximum concentration. Investigation of the role of cell density on product formation (Fig.
Bioreactor growths were conducted in a New Brunswick BioFlo110 fermentor, set up according the manufacturer's instructions. Under our settings, the cultures were stirred at 300 rpm, their temperature maintained at 20 °C, pH held at 7.2 (using 5 M KOH and 5 M HCL reservoirs connected to the A and B pumps), chemically resistant tubing was used, and air flow was maintained at 4 lpm using triple micron filtered air. Pulse feeding supplements were introduced through the feeding septa, and cell density and product output monitored, as well as pH verified, by intermittent sample removal for subsequent measurement.
S. aureus strains were grown in tryptic soy broth (TSB) at 30°C or 37°C with aeration, unless otherwise indicated. The medium was supplemented with erythromycin (5 μg/ml for chromosomal insertions and 10 μg/ml for plasmids), chloramphenicol (5 μg/ml for chromosomal insertions, 10 μg/ml for plasmid), spectinomycin (100 μg/ml), kanamycin and neomycin (25 μg/ml each), 5-bromo-4-chloro-3-indolyl ß-D-galactopyranoside (X-Gal, 250 μg/ml), or anhydrotetracycline (aTc, 100 ng/ml unless otherwise indicated). B. subtilis strains were derived from the prototrophic strain PY79 [40 (link)]. Cells were grown in Luria Broth (LB) or Casein Hydrolysate (CH) medium [41 ] at 37°C with aeration, unless otherwise indicated. The medium was supplemented with tetracycline (10 μg/ml), spectinomycin (100 μg/ml), kanamycin 10 μg/ml), chloramphenicol (5 μg/ml), xylose (0.5% w/v) or isopropyl ß-d-thiogalactopyranoside (IPTG, 0.5 mM). Lists of oligonucleotide primers, strains, plasmids, and descriptions of their construction can be found in Supplemental Material.
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5-bromo-4-chloro-3-indolyl beta-galactoside
anhydrotetracycline
casein hydrolysate
Cells
Chloramphenicol
Chromosomes, Human, Pair 5
Erythromycin
Galactose
Insertion Mutation
Isopropyl Thiogalactoside
Kanamycin
Neomycin
Oligonucleotide Primers
Plasmids
Spectinomycin
Staphylococcus aureus
Strains
Tetracycline
tryptic soy broth
Xylose
Most recents protocols related to «Casein hydrolysate»
The experiments were carried out in the tissue culture laboratory for Date Palm Research Centre at Basrah University, Basrah, Iraq.
Young offshoots (2-3 years old) of date palm cv. Barhee were detached from the parent palm. Outer leaves and fibrous tissues at their bases were removed gradually until the shoot tip zone was exposed. Sterilization of explants was performed using 70% ethanol for 1 min and 2.5% sodium hypochlorite for 20 min. Explants were then rinsed three times with sterile distilled water. For callus induction, explants were cultured on the MS basal medium (Murashige and Skoog, 1962) with the addition of 3.0 mg l -1 6-(dimethylallyl amino purine) (2iP), 30 mg l -1 of 1-naphthalene acetic acid (NAA), 30 g l -1 of sucrose, 2.0 g l -1 of activated charcoal, and solidified with 6.0 g l -1 agar-agar. The cultures were transferred to fresh media, with the same composition every 6 weeks until the callus began to initiate. All cultures were incubated in a culture room under darkness at 27 ± 2 °C for 180 days to initiate callus.
To study the effects of dicamba (DIC) and casein hydrolysate (CH), supplementation of these compounds at various concentrations in the growth media was assessed. The induced callus was separated from the apical buds, weighed, and cultured (100 mg per jar) on MS medium, with the addition of growth regulators NAA at 6.0 mg l -1 and 2iP at 2.0 mg l -1 . To study the effects of dicamba (DIC) and casein hydrolysate (CH) on callus growth, supplementation of these compounds at different concentrations in the growth medium was assessed. MS medium was supplemented with 0.0, 2.0, 4.0, and 6.0 mg l -1 DIC (Sigma-Aldrich, St. Louis, MO) or 0.0, 0.5, and 1.0 g l -1 CH, or added together at 4.0 mg l -1 DIC + 0.5 g l -1 CH, and 4.0 mg l -1 DIC + 1.0 g l -1 CH. The pH of the medium was adjusted to 5.7-5.8 before the addition of agar. Media were dispensed into culture containers and autoclaved at 121 °C and 1.04 kg cm -2 for 20 min. The cultures were kept in a culture room at 27 ± 2 °C with 16 h light and irradiance of 13.5 μmol m -2 s -1 provided by cool white fluorescent lamps at the light intensity of 2,000 lux.
For differentiation and multiplication, the callus on growth media was divided and subcultured on differentiation and multiplication media supplemented as mentioned above, except for the plant growth regulators 1 mg l -1 (NAA), 0.5 mg l -1 6-benzyladenine (BA), and 0.5 mg l -1 kinetin (K). It was also supplemented with the same DIC and CH concentrations to study their effects on buds multiplication and some changes in phytochemicals properties. The cultures were maintained under room temperature 27 ± 2 °С, with a photoperiod of 16/8 h day/night. The light intensity was 2,000 lux provided by cool white fluorescent lamps. There were 18 replicates of each treatment. The percentage of bud regeneration and bud number per jar were recorded at 12 weeks from the inoculation of callus on the media.
Young offshoots (2-3 years old) of date palm cv. Barhee were detached from the parent palm. Outer leaves and fibrous tissues at their bases were removed gradually until the shoot tip zone was exposed. Sterilization of explants was performed using 70% ethanol for 1 min and 2.5% sodium hypochlorite for 20 min. Explants were then rinsed three times with sterile distilled water. For callus induction, explants were cultured on the MS basal medium (Murashige and Skoog, 1962) with the addition of 3.0 mg l -1 6-(dimethylallyl amino purine) (2iP), 30 mg l -1 of 1-naphthalene acetic acid (NAA), 30 g l -1 of sucrose, 2.0 g l -1 of activated charcoal, and solidified with 6.0 g l -1 agar-agar. The cultures were transferred to fresh media, with the same composition every 6 weeks until the callus began to initiate. All cultures were incubated in a culture room under darkness at 27 ± 2 °C for 180 days to initiate callus.
To study the effects of dicamba (DIC) and casein hydrolysate (CH), supplementation of these compounds at various concentrations in the growth media was assessed. The induced callus was separated from the apical buds, weighed, and cultured (100 mg per jar) on MS medium, with the addition of growth regulators NAA at 6.0 mg l -1 and 2iP at 2.0 mg l -1 . To study the effects of dicamba (DIC) and casein hydrolysate (CH) on callus growth, supplementation of these compounds at different concentrations in the growth medium was assessed. MS medium was supplemented with 0.0, 2.0, 4.0, and 6.0 mg l -1 DIC (Sigma-Aldrich, St. Louis, MO) or 0.0, 0.5, and 1.0 g l -1 CH, or added together at 4.0 mg l -1 DIC + 0.5 g l -1 CH, and 4.0 mg l -1 DIC + 1.0 g l -1 CH. The pH of the medium was adjusted to 5.7-5.8 before the addition of agar. Media were dispensed into culture containers and autoclaved at 121 °C and 1.04 kg cm -2 for 20 min. The cultures were kept in a culture room at 27 ± 2 °C with 16 h light and irradiance of 13.5 μmol m -2 s -1 provided by cool white fluorescent lamps at the light intensity of 2,000 lux.
For differentiation and multiplication, the callus on growth media was divided and subcultured on differentiation and multiplication media supplemented as mentioned above, except for the plant growth regulators 1 mg l -1 (NAA), 0.5 mg l -1 6-benzyladenine (BA), and 0.5 mg l -1 kinetin (K). It was also supplemented with the same DIC and CH concentrations to study their effects on buds multiplication and some changes in phytochemicals properties. The cultures were maintained under room temperature 27 ± 2 °С, with a photoperiod of 16/8 h day/night. The light intensity was 2,000 lux provided by cool white fluorescent lamps. There were 18 replicates of each treatment. The percentage of bud regeneration and bud number per jar were recorded at 12 weeks from the inoculation of callus on the media.
The effect of the treatment was evaluated following a well-established protocol for the C. jejuni cecal model, as outlined by Olson et al. and Feye et al. [16 (link),19 ]. Cecal contents were collected from six individual birds within aseptic chambers, then weighed and diluted at a ratio of 1:3000 by mixing 0.1 g of cecal content with 900 μL of ADS solution. This resuspended cecal material was further diluted by adding 1 mL of it to 299 mL of ADS for each respective cecum sample. A 36 mL aliquot of this diluted cecal content was subsequently transferred into each serum bottle, with or without the addition of 0.4 g of ground chicken feed (S1 Table ). To create an appropriate environment, the cultures were covered with aluminum foil and placed in Advanced Anoxomat™ III containers (Advanced Instruments, Norwood, MA, USA), where a microaerobic atmosphere was established, containing 5% O2, 10% CO2, and 85% N2. Incubation was carried out at 42°C with continuous agitation at 150 revolutions per minute (RPM) for 24 hours. This step served as a pre-adaptation to acclimate the naturally occurring microbiota in poultry ceca to the new environment, following the method described previously [18 (link)–20 (link)]. On the following day, the serum bottles were returned to the anaerobic chamber.
After 24 h of pre-adaption, the serum bottles were moved back into the anaerobic chamber where 4 ml of a freshly prepared 106 cells/mL C. jejuni culture resuspended in ADS was added to all bottles except the non-inoculated treatment (NI) for a final concentration of 105 CFU/mL. For specific treatments involving casein supplements, 0.1 g of the appropriate vitamin free-casein type (intact (Envigo®, Madison, WI, USA), enzyme hydrolyzed (MilliporeSigma, Burlington, MA, USA), or acid hydrolysate of casein (MilliporeSigma, Burlington, MA, USA) was added to the serum bottles. The study included five distinct treatments: non-inoculated (NI), inoculated with C. jejuni (IN), and three supplemented groups, intact casein (IC), enzyme hydrolysate (EH), acid hydrolysate (AH), that were all inoculated with C. jejuni. We opted not to include additional controls for supplemented groups without Campylobacter, instead using non-inoculated and inoculated treatments without casein supplementations for comparative analyses with the casein-enriched groups. Because Campylobacter naturally inhabits poultry cecal compartments [21 (link)], we confirmed the absence of culturable Campylobacter in the NI group and confirm the inocula, the 0-hour samples were plated on mCCDA [22 (link)]. At 0, 24, and 48 h post-inoculation, duplicate 1 mL samples were taken for microbiome sequencing and metabolomic analysis. The samples were flash-frozen in liquid nitrogen and stored at -80°C until processing. An overview of the procedure is described inFig 1 .
After 24 h of pre-adaption, the serum bottles were moved back into the anaerobic chamber where 4 ml of a freshly prepared 106 cells/mL C. jejuni culture resuspended in ADS was added to all bottles except the non-inoculated treatment (NI) for a final concentration of 105 CFU/mL. For specific treatments involving casein supplements, 0.1 g of the appropriate vitamin free-casein type (intact (Envigo®, Madison, WI, USA), enzyme hydrolyzed (MilliporeSigma, Burlington, MA, USA), or acid hydrolysate of casein (MilliporeSigma, Burlington, MA, USA) was added to the serum bottles. The study included five distinct treatments: non-inoculated (NI), inoculated with C. jejuni (IN), and three supplemented groups, intact casein (IC), enzyme hydrolysate (EH), acid hydrolysate (AH), that were all inoculated with C. jejuni. We opted not to include additional controls for supplemented groups without Campylobacter, instead using non-inoculated and inoculated treatments without casein supplementations for comparative analyses with the casein-enriched groups. Because Campylobacter naturally inhabits poultry cecal compartments [21 (link)], we confirmed the absence of culturable Campylobacter in the NI group and confirm the inocula, the 0-hour samples were plated on mCCDA [22 (link)]. At 0, 24, and 48 h post-inoculation, duplicate 1 mL samples were taken for microbiome sequencing and metabolomic analysis. The samples were flash-frozen in liquid nitrogen and stored at -80°C until processing. An overview of the procedure is described in
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Basing on LB fermentation medium, single factor at a time was used for optimization. The BcMFAse activity was analyzed after incubating for 48 h. Firstly, the basal medium (0.5% yeast extract and 1% NaCl) were supplemented with 1%-3% tryptone to determine the optimal concentration of tryptone. Secondly, the basal medium (the optimal concentration of tryptone and 1% NaCl) were supplemented with 0.3%-1.5% yeast extract to determine the optimal concentration of yeast extract. Finally, according to previous reports, casein hydrolysate helped improving the production of protein (Pedersen et was considered adding to the fermentation medium. The basal medium (optimal concentration of tryptone, optimal concentration of yeast extract and 1% NaCl) were supplemented with 0.2%-1% casein hydrolysate to determine the optimal concentration of casein hydrolysate.
The preparation of BM169 refers to studies of S. mansoni [12 (link), 13 (link)], using a liquid BME (Basal Medium Eagle) (Gibco) as a substitute for the powder BME in this study. We produced AB169 by adding 200 μM ascorbic acid (Sigma-Aldrich) and 0.2% V/V bovine washed red bloods cells (10% suspension; Hongquan Bio, Guangzhou, China) into BM169. AB169 (1640) was made by substituting the BME with RPMI-1640 (Gibco) in AB169. M-AB169 (1640) was obtained by replacing half the amount of the lactalbumin hydrolysate (Sigma-Aldrich) in AB169 (1640) with casein hydrolysate (Merck Millipore). M-BM169 (1640) was formulated by replacing BME with RPMI-1640 and incorporating casein hydrolysate in place of half of the lactalbumin hydrolysate present in the original BM169. Detailed compositions of the media are provided in Additional file 7 : Table S1.
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The bacterial strain used in the study, C. violaceum 30191, was obtained from the Leibniz-Institute DSMZ-German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany). It was stored in 8% DMSO at −80 °C. Before each experiment, the strain was inoculated in Luria-Bertani Broth (LB)—1% Casein enzymic hydrolysate, 0.5% yeast extract, 1% NaCl, (HiMedia, Modautal, Germany), and maintained at 4 °C on Luria-Bertani Agar (LA)—1% Casein enzymic hydrolysate, 0.5% yeast extract, 1% NaCl, 0.15% agar (HiMedia, Modautal, Germany) slants. As a source of bacterial inoculum, before each experiment, the strain was incubated in LB, without shaking at 30 °C for 24 h.
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Casein hydrolysate is a mixture of amino acids and small peptides derived from the enzymatic hydrolysis of casein, a protein found in milk. It serves as a nutrient source for cell culture media and microbial growth applications.
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
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Yeast extract is a versatile laboratory product derived from the autolysis of baker's yeast. It contains a rich source of amino acids, vitamins, and other essential nutrients that are beneficial for the cultivation and growth of microorganisms. Yeast extract is commonly used as a nutrient supplement in microbial culture media for a variety of applications, including fermentation processes, cell culture, and the production of various biological products.
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Starch is a carbohydrate-based laboratory reagent used in various testing and analytical procedures. It serves as a general indicator for the presence of iodine, which forms a characteristic blue-black complex when combined with starch. The core function of starch is to provide a visual detection method for iodine, without further interpretation or extrapolation on its intended use.
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The vacuum filtration system is a laboratory equipment designed to facilitate the process of separating solids from liquids. It utilizes a vacuum to draw the liquid through a filter, leaving the solid particles behind. The system is composed of a filtration flask, a vacuum pump, and a filter holder that can accommodate various filter media.
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The NanoDrop ND-1000 spectrophotometer is a compact and easy-to-use instrument designed for the quantification of small volume samples. It utilizes a proprietary sample retention system to measure the absorbance of samples as small as 1 μL, making it suitable for a wide range of applications in molecular biology, genomics, and biochemistry.
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Bovine serum albumin (BSA) is a common laboratory reagent derived from bovine blood plasma. It is a protein that serves as a stabilizer and blocking agent in various biochemical and immunological applications. BSA is widely used to maintain the activity and solubility of enzymes, proteins, and other biomolecules in experimental settings.
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Trypsin is a serine protease enzyme that is commonly used in cell biology and biochemistry laboratories. Its primary function is to facilitate the dissociation and disaggregation of adherent cells, allowing for the passive release of cells from a surface or substrate. Trypsin is widely utilized in various cell culture applications, such as subculturing and passaging of adherent cell lines.
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Casein hydrolysate is a protein product derived from the enzymatic hydrolysis of casein, a milk protein. It provides a source of amino acids and peptides. The specific composition and properties may vary depending on the manufacturing process.
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Ascorbic acid is a chemical compound commonly known as Vitamin C. It is a water-soluble vitamin that plays a role in various physiological processes. As a laboratory product, ascorbic acid is used as a reducing agent, antioxidant, and pH regulator in various applications.
More about "Casein hydrolysate"
Casein hydrolysate, also known as milk protein hydrolysate or caseinolytic digest, is a nutrient-rich product derived from the enzymatic breakdown of casein, the primary protein found in milk.
This complex mixture contains a variety of amino acids, peptides, and other bioactive compounds that have diverse applications in the food, pharmaceutical, and nutraceutical industries.
Casein hydrolysate is commonly used as a dietary supplement, a source of protein in sports nutrition products, and as an ingredient in specialized medical foods.
Its unique composition and properties make it a valuable tool for researchers exploring its potential health benefits, such as its ability to support muscle recovery, bone health, and immune function.
Optimizing the production and characterization of casein hydrolysate is an important area of study.
Researchers can utilize PubCompare.ai, an AI-powered platform, to easily locate and compare relevant protocols from the scientific literature, preprints, and patents.
This can help identify the best methods and products for their casein hydrolysate research, enhancing reproducibility and accuracy.
In addition to casein hydrolysate, other related compounds like fetal bovine serum (FBS), yeast extract, and starch may also be of interest for researchers in this field.
Techniques such as vacuum filtration systems and spectrophotometric analysis using a NanoDrop ND-1000 can be employed to purify and characterize these materials.
Furthermore, bovine serum albumin, trypsin, and ascorbic acid are commonly used in experiments involving casein hydrolysate and related compounds.
By leveraging the insights and tools provided by PubCompare.ai, researchers can optimize their casein hydrolysate research and unlock its full potential in the food, pharmaceutical, and nutraceutical industries.
This complex mixture contains a variety of amino acids, peptides, and other bioactive compounds that have diverse applications in the food, pharmaceutical, and nutraceutical industries.
Casein hydrolysate is commonly used as a dietary supplement, a source of protein in sports nutrition products, and as an ingredient in specialized medical foods.
Its unique composition and properties make it a valuable tool for researchers exploring its potential health benefits, such as its ability to support muscle recovery, bone health, and immune function.
Optimizing the production and characterization of casein hydrolysate is an important area of study.
Researchers can utilize PubCompare.ai, an AI-powered platform, to easily locate and compare relevant protocols from the scientific literature, preprints, and patents.
This can help identify the best methods and products for their casein hydrolysate research, enhancing reproducibility and accuracy.
In addition to casein hydrolysate, other related compounds like fetal bovine serum (FBS), yeast extract, and starch may also be of interest for researchers in this field.
Techniques such as vacuum filtration systems and spectrophotometric analysis using a NanoDrop ND-1000 can be employed to purify and characterize these materials.
Furthermore, bovine serum albumin, trypsin, and ascorbic acid are commonly used in experiments involving casein hydrolysate and related compounds.
By leveraging the insights and tools provided by PubCompare.ai, researchers can optimize their casein hydrolysate research and unlock its full potential in the food, pharmaceutical, and nutraceutical industries.