ABSTRACT
Folate, a vital water-soluble vitamin (B9), requires specific attention as its recommended daily intake frequently is not reached in countries without mandatory fortification. In this regard, biofortification with microorganisms like Bifidobacterium and Streptococcus offers a compelling approach for enhancing food with natural folates. A randomized, nonblinded, and monocentric human pilot study is conducted to assess the bioavailability of a folate-biofortified fermented whey beverage, comprising 3 intervention days and a controlled replenishment phase before and during the assay. Folate plasma concentration (5-CH3-H4folate) is determined using a stable isotope dilution assay and LC-MS/MS detection. Biokinetic parameters (cmax and tmax) are determined, and areas under the curve (AUC) normalized to the basal folate plasma concentration are calculated. An average bioavailability of 17.1% in relation to the 5-CH3-H4folate supplement, ranging from 0% to 39.8%, is obtained. These results reiterate the significance of additional research into folate bioavailability in general and dairy products. Further investigations are warranted into folate-binding proteins (FBP) and other potential limiting factors within the food and individual factors. In summary, biofortification via fermentation emerges as a promising avenue for enhancing the natural folate content in dairy and other food products.
Subject(s)
Folic Acid , Humans , Folic Acid/pharmacokinetics , Folic Acid/administration & dosage , Folic Acid/blood , Adult , Female , Male , Whey/chemistry , Food, Fortified , Pilot Projects , Fermentation , Biological Availability , Young Adult , Biofortification/methods , Tetrahydrofolates/pharmacokinetics , Middle Aged , Beverages/analysisABSTRACT
This study explores a sustainable approach for synthesizing silver nanocomposites (AgNCs) with enhanced antimicrobial and bioactivity using safe Lactobacillus strains and a whey-based medium (WBM). WBM effectively supported the growth of Lactobacillus delbrueckii and Lactobacillus acidophilus, triggering a stress response that led to AgNCs formation. The synthesized AgNCs were characterized using advanced spectroscopic and imaging techniques such as UVâvisible, Fourier transform infrared (FT-IR) spectroscopy, transmission electron (TEM), and scanning electron microscopy with energy dispersive X-ray analysis (SEM-Edx). Lb acidophilus-synthesized AgNCs in WBM (had DLS size average 817.2-974.3 ± PDI = 0.441 nm with an average of metal core size 13.32 ± 3.55 nm) exhibited significant antimicrobial activity against a broad spectrum of pathogens, including bacteria such as Escherichia coli (16.47 ± 2.19 nm), Bacillus cereus (15.31 ± 0.43 nm), Clostridium perfringens (25.95 ± 0.03 mm), Enterococcus faecalis (32.34 ± 0.07 mm), Listeria monocytogenes (23.33 ± 0.05 mm), methicillin-resistant Staphylococcus aureus (MRSA) (13.20 ± 1.76 mm), and filamentous fungi such as Aspergillus brasiliensis (33.46 ± 0.01 mm). In addition, Lb acidophilus-synthesized AgNCs in WBM exhibit remarkable free radical scavenging abilities, suggesting their potential as bioavailable antioxidants. These findings highlight the dual functionality of these biogenic AgNCs, making them promising candidates for applications in both medicine and nutrition.
Subject(s)
Microbial Sensitivity Tests , Nanocomposites , Silver , Whey , Nanocomposites/chemistry , Silver/chemistry , Silver/pharmacology , Whey/chemistry , Whey/metabolism , Lactobacillus acidophilus/drug effects , Lactobacillus acidophilus/metabolism , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/biosynthesis , Metal Nanoparticles/chemistry , Lactobacillus/metabolism , Anti-Infective Agents/pharmacology , Anti-Infective Agents/chemistry , Spectroscopy, Fourier Transform InfraredABSTRACT
The probiotic properties of twenty-five lactic acid bacteria (LAB) isolated from human breast milk were investigated considering their resistance to gastrointestinal conditions and proteolytic activity. Seven LAB were identified and assessed for auto- and co-aggregation capacity, antibiotic resistance, and behavior during in vitro gastrointestinal digestion. Three Lacticaseibacillus strains were further evaluated for antifungal activity, metabolite production (HPLC-Q-TOF-MS/MS and GC-MS/MS) and proteolytic profiles (SDS-PAGE and HPLC-DAD) in fermented milk, whey, and soy beverage. All strains resisted in vitro gastrointestinal digestion with viable counts higher than 7.9 log10 CFU mL-1 after the colonic phase. Remarkable proteolytic activity was observed for 18/25 strains. Bacterial auto- and co-aggregation of 7 selected strains reached values up to 23 and 20%, respectively. L. rhamnosus B5H2, L. rhamnosus B9H2 and L. paracasei B10L2 inhibited P. verrucosum, F. verticillioides and F. graminearum fungal growth, highlighting L. rhamnosus B5H2. Several metabolites were identified, including antifungal compounds such as phenylacetic acid and 3-phenyllactic acid, and volatile organic compounds produced in fermented milk, whey, and soy beverage. SDS-PAGE demonstrated bacterial hydrolysis of the main milk (caseins) and soy (glycines and beta-conglycines) proteins, with no apparent hydrolysis of whey proteins. However, HPLC-DAD revealed alpha-lactoglobulin reduction up to 82% and 54% in milk and whey, respectively, with L. rhamnosus B5H2 showing the highest proteolytic activity. Overall, the three selected Lacticaseibacillus strains demonstrated probiotic capacity highlighting L. rhamnosus B5H2 with remarkable potential for generating bioactive metabolites and peptides which are capable of promoting human health.
Subject(s)
Dietary Supplements , Lactobacillales , Milk, Human , Probiotics , Humans , Milk, Human/chemistry , Female , Lactobacillales/metabolism , Lactobacillales/isolation & purification , Fermentation , Whey/microbiology , Whey/chemistry , Phenylacetates/metabolism , Antifungal Agents/pharmacology , Antifungal Agents/metabolism , Cultured Milk Products/microbiology , LactatesABSTRACT
This study is focused on fractionation of insulin-like growth factor I (IGF-I) and transforming growth factor-ß2 (TGF-ß2) using a new electro-based membrane process calledelectrodialysis with filtration membranes (EDFM). Before EDFM, different pretreatments were tested, and four pH conditions (4.25, 3.85, 3.45, and 3.05) were used during EDFM. It was demonstrated that a 1:1 dilution of defatted colostrum with deionized water to decrease mineral content followed by the preconcentration of GFs by UF is necessary and allow for these compounds to migrate to the recovery compartment during EDFM. MS analyses confirmed the migration, in low quantity, of only α-lactalbumin (α-la) and ß-lactoglobulin (ß-lg) from serocolostrum to the recovery compartment during EDFM. Consequently, the ratio of GFs to total protein in recovery compartment compared to that of feed serocolostrum solution was 60× higher at pH value 3.05, the optimal pH favoring the migration of IGF-I and TGF-ß2. Finally, these optimal conditions were tested on acid whey to also demonstrate the feasibility of the proposed process on one of the main by-products of the cheese industry; the ratio of GFs to total protein was 2.7× higher in recovery compartment than in feed acid whey solution, and only α-la migrated. The technology of GF enrichment for different dairy solutions by combining ultrafiltration and electrodialysis technologies was proposed for the first time.
Subject(s)
Dialysis , Filtration , Dialysis/methods , Filtration/methods , Insulin-Like Growth Factor I/analysis , Hydrogen-Ion Concentration , Membranes, Artificial , Dairy Products/analysis , Animals , Colostrum/chemistry , Cattle , Whey/chemistry , Lactoglobulins/chemistry , Lactoglobulins/analysis , Lactalbumin/chemistry , Lactalbumin/analysisABSTRACT
The constantly diverse demand scenarios for rapid on-site analysis have put forward high requirements for developing low-cost and user-friendly visual detection methods. Therefore, developing a visual detection method with simple operation and intuitive results has important practical value in the field of analysis and detection, but it is also challenging. In this work, we propose a microsyringe-assisted visual volume detection method based on phase separation, and apply it to analyze the milk-clotting activity of chymosin. Chymosin can cause phase separation of milk with whey in the mobile phase and curd in the gel state. The network structures of casein in curd can trap water molecules, resulting in separation of whey from curd gradually. Therefore, the analysis of chymosin milk-clotting activity can be realized according to the volume of whey measured using a portable microsyringe. This method shows a good linear correlation when the concentration of chymosin ranges from 1.02 U L-1 to 1020 U L-1 and the limit of detection of this method for chymosin is calculated to be 0.03 U mL-1. This work successfully realizes the visual analysis of chymosin milk-clotting activity based on the enzyme-triggered phase separation. It also shows great promise to be applied in other phase separation-based detection systems with the advantages of high accuracy, great portability and user-friendliness.
Subject(s)
Chymosin , Milk , Chymosin/chemistry , Chymosin/metabolism , Milk/chemistry , Animals , Whey/chemistry , Caseins/analysis , Caseins/chemistry , Phase SeparationABSTRACT
The structural and functional properties of whey-quercetin and whey hydrolysate-quercetin conjugates synthesized using alkaline and free radical-mediated methods (AM and FRM) coupled with sonication were studied. FTIR showed new peaks at 3000-3500 cm-1 (N-H stretching regions) and the 1000-1100 cm-1 region with the conjugates. Conjugation increased the random coils and α-helix content while decreasing the ß-sheets and turns. It also increased the particle size and surface hydrophobicity which was significantly (p < 0.05) higher in AM than FRM conjugates. AM conjugates had higher radical scavenging activity but lower quercetin content than FRM conjugates. Overall, the functional properties of whey-quercetin conjugates were better than whey hydrolysate-quercetin conjugates. However, hydrolysate conjugates had significantly higher denaturation temperatures irrespective of the method of production. Sonication improved the radical scavenging activity and quercetin content of FRM conjugates while it decreased both for AM conjugates. This study suggested that whey-quercetin conjugates generally had better quality than whey hydrolysate conjugates and sonication tended to further improve these properties. This study highlights the potential for using camel whey or whey hydrolysate-quercetin conjugates to enhance the functional properties of food products in the food industry.
Subject(s)
Camelus , Hydrophobic and Hydrophilic Interactions , Quercetin , Sonication , Quercetin/chemistry , Animals , Protein Hydrolysates/chemistry , Whey/chemistry , Antioxidants/chemistry , Whey Proteins/chemistry , Free Radical Scavengers/chemistry , Spectroscopy, Fourier Transform Infrared , Free Radicals/chemistry , Particle Size , Hydrogen-Ion ConcentrationABSTRACT
Recovering valuable active substances from the by-products of agricultural processing is a crucial concern for scientific researchers. This paper focuses on the enrichment of soybean trypsin inhibitor (STI) from soybean whey wastewater using either ammonium sulfate salting or ethanol precipitation, and discusses their physicochemical properties. The results show that at a 60% ethanol content, the yield of STI was 3.983 mg/mL, whereas the yield was 3.833 mg/mL at 60% ammonium sulfate saturation. The inhibitory activity of STI obtained by ammonium sulfate salting out (A-STI) was higher than that obtained by ethanol precipitation (E-STI). A-STI exhibited better solubility than E-STI at specific temperatures and pH levels, as confirmed by turbidity and surface hydrophobicity measurements. Thermal characterization revealed that both A-STI and E-STI showed thermal transition temperatures above 90 °C. Scanning electron microscopy demonstrated that A-STI had a smooth surface with fewer pores, while E-STI had a rough surface with more pores. In conclusion, there was no significant difference in the yield of A-STI and E-STI (p < 0.05); however, the physicochemical properties of A-STI were superior to those of E-STI, making it more suitable for further processing and utilization. This study provides a theoretical reference for the enrichment of STI from soybean whey wastewater.
Subject(s)
Glycine max , Trypsin Inhibitors , Wastewater , Whey , Glycine max/chemistry , Wastewater/chemistry , Whey/chemistry , Trypsin Inhibitors/chemistry , Trypsin Inhibitors/isolation & purification , Ammonium Sulfate/chemistry , Chemical Precipitation , Hydrogen-Ion Concentration , Solubility , Hydrophobic and Hydrophilic Interactions , TemperatureABSTRACT
In the field of biotechnology, the utilization of agro-industrial waste for generating high-value products, such as microbial biomass and enzymes, holds significant importance. This study aimed to produce recombinant α-amylase from Anoxybacillus karvacharensis strain K1, utilizing whey as an useful growth medium. The purified hexahistidine-tagged α-amylase exhibited remarkable homogeneity, boasting a specific activity of 1069.2 U mg-1. The enzyme displayed its peak activity at 55 °C and pH 6.5, retaining approximately 70% of its activity even after 3 h of incubation at 55 °C. Its molecular weight, as determined via SDS-PAGE, was approximately 69 kDa. The α-amylase demonstrated high activity against wheat starch (1648.8 ± 16.8 U mg-1) while exhibiting comparatively lower activity towards cyclodextrins and amylose (≤ 200.2 ± 16.2 U mg-1). It exhibited exceptional tolerance to salt, withstanding concentrations of up to 2.5 M. Interestingly, metal ions and detergents such as sodium dodecyl sulfate (SDS), Triton 100, Triton 40, and Tween 80, 5,5'-dithio-bis-[2-nitrobenzoic acid (DNTB), ß-mercaptoethanol (ME), and dithiothreitol (DTT) had no significant inhibitory effect on the enzyme's activity, and the presence of CaCl2 (2 mM) even led to a slight activation of the recombinant enzyme (1.4 times). The Michaelis constant (Km) and maximum reaction rate (Vmax), were determined using soluble starch as a substrate, yielding values of 1.2 ± 0.19 mg mL-1 and 1580.3 ± 183.7 µmol mg-1 protein min-1, respectively. Notably, the most favorable conditions for biomass and recombinant α-amylase production were achieved through the treatment of acid whey with ß-glucosidase for 24 h.
Subject(s)
Anoxybacillus , Detergents , Whey , alpha-Amylases , alpha-Amylases/metabolism , alpha-Amylases/chemistry , Whey/metabolism , Whey/chemistry , Anoxybacillus/enzymology , Anoxybacillus/genetics , Detergents/chemistry , Hydrogen-Ion Concentration , Enzyme Stability , Recombinant Proteins/metabolism , Recombinant Proteins/isolation & purification , Recombinant Proteins/genetics , Recombinant Proteins/chemistry , Starch/metabolism , Starch/chemistry , TemperatureABSTRACT
Whey is a byproduct of dairy industries, the aqueous portion which separates from cheese during the coagulation of milk. It represents approximately 85-95% of milk's volume and retains much of its nutrients, including functional proteins and peptides, lipids, lactose, minerals, and vitamins. Due to its composition, mainly proteins and lactose, it can be considered a raw material for value-added products. Whey-derived products are often used to supplement food, as they have shown several physiological effects on the body. Whey protein hydrolysates are reported to have different activities, including antihypertensive, antioxidant, antithrombotic, opioid, antimicrobial, cytomodulatory, and immuno-modulatory. On the other hand, galactooligosaccharides obtained from lactose can be used as prebiotic for beneficial microorganisms for the human gastrointestinal tract. All these compounds can be obtained through physicochemical, microbial, or enzymatic treatments. Particularly, enzymatic processes have the advantage of being highly selective, more stable than chemical transformations, and less polluting, making that the global enzyme market grow at accelerated rates. The sources and different products associated with the most used enzymes are particularly highlighted in this review. Moreover, we discuss metagenomics as a tool to identify novel proteolytic enzymes, from both cultivable and uncultivable microorganisms, which are expected to have new interesting activities. Finally enzymes for the transformation of whey sugar are reviewed. In this sense, carbozymes with ß-galactosidase activity are capable of lactose hydrolysis, to obtain free monomers, and transgalactosylation for prebiotics production. KEY POINTS: ⢠Whey can be used to obtain value-added products efficiently through enzymatic treatments ⢠Proteases transform whey proteins into biopeptides with physiological activities ⢠Lactose can be transformed into prebiotic compounds using ß-galactosidases.
Subject(s)
Protein Hydrolysates , Whey Proteins , Whey Proteins/metabolism , Protein Hydrolysates/metabolism , Protein Hydrolysates/chemistry , Prebiotics , Humans , Whey/chemistry , Whey/metabolism , Lactose/metabolism , beta-Galactosidase/metabolism , beta-Galactosidase/geneticsABSTRACT
Kars Kashar cheese is an artisanal pasta-filata type cheese and geographically marked in Eastern Anatolia of Turkey. The aims of this research were to determine for the first time thermophilic lactic acid bacteria (LAB) of Kars Kashar cheese and characterize the technological properties of obtained isolates. In our research, a number of 15 samples of whey were collected from the different villages in Kars. These samples were incubated at 45 °C and used as the source material for isolating thermophilic LAB. A total of 250 colonies were isolated from thermophilic whey, and 217 of them were determined to be presumptive LAB based on their Gram staining and catalase test. A total of 170 isolates were characterized by their phenotypic properties and identified using the MALDI-TOF mass spectrometry method. Phenotypic identification of isolates displayed that Enterococcus and Lactobacillus were the predominant microbiota. According to MALDI-TOF MS identification, 89 isolates were identified as Enterococcus (52.35%), 57 isolates as Lactobacillus (33.53%), 23 isolates as Streptococcus (13.53%), and one isolate as Lactococcus (0.59%). All thermophilic LAB isolates were successfully identified to the species level and it has been observed that MALDI-TOF MS can be successfully used for the identification of selected LAB. The acidification and proteolytic activities of the isolated thermophilic LAB were examined, and the isolates designated for use as starter cultures were also genotypically defined.
Subject(s)
Cheese , Lactobacillales , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Cheese/microbiology , Lactobacillales/isolation & purification , Lactobacillales/classification , Lactobacillales/genetics , Lactobacillales/metabolism , Whey/microbiology , Whey/chemistry , Food Microbiology , Turkey , Lactobacillus/isolation & purification , Lactobacillus/genetics , Lactobacillus/classification , Lactobacillus/metabolism , Enterococcus/isolation & purification , Enterococcus/classification , Enterococcus/genetics , Enterococcus/metabolismABSTRACT
Using whey, a by-product of the cheese-making process, is important for maximizing resource efficiency and promoting sustainable practices in the food industry. Reusing whey can help minimize environmental impact and produce bio-preservatives for foods with high bacterial loads, such as Mexican-style fresh cheeses. This research aims to evaluate the antimicrobial and physicochemical effect of CFS from Lactobacillus casei 21/1 produced in a conventional culture medium (MRS broth) and another medium using whey (WB medium) when applied in Mexican-style fresh cheese inoculated with several indicator bacteria (Escherichia coli, Salmonella enterica serovar Typhimurium, Staphylococcus aureus, and Listeria monocytogenes). The CFSs (MRS or WB) were characterized for organic acids concentration, pH, and titratable acidity. By surface spreading, CFSs were tested on indicator bacteria inoculated in fresh cheese. Microbial counts were performed on inoculated cheeses during and after seven days of storage at 4 ± 1.0 °C. Moreover, pH and color were determined in cheeses with CFS treatment. Lactic and acetic acid were identified as the primary antimicrobial metabolites produced by the Lb. casei 21/1 fermentation in the food application. A longer storage time (7 days) led to significant reductions (p < 0.05) in the microbial population of the indicator bacteria inoculated in the cheese when it was treated with the CFSs (MRS or WB). S. enterica serovar Typhimurium was the most sensitive bacteria, decreasing 1.60 ± 0.04 log10 CFU/g with MRS-CFS, whereas WB-CFS reduced the microbial population of L. monocytogenes to 1.67 log10 CFU/g. E. coli and S. aureus were the most resistant at the end of storage. The cheese's pH with CFSs (MRS or WB) showed a significant reduction (p < 0.05) after CFS treatment, while the application of WB-CFS did not show greater differences in color (ΔE) compared with MRS-CFS. This study highlights the potential of CFS from Lb. casei 21/1 in the WB medium as an ecological bio-preservative for Mexican-style fresh cheese, aligning with the objectives of sustainable food production and guaranteeing food safety.
Subject(s)
Cheese , Lacticaseibacillus casei , Whey , Cheese/microbiology , Cheese/analysis , Lacticaseibacillus casei/metabolism , Whey/chemistry , Whey/microbiology , Food Microbiology , Hydrogen-Ion Concentration , Food Preservation/methods , Mexico , FermentationABSTRACT
Buttermilk is a potential material for the production of a milk fat globule membrane (MFGM) and can be mainly classified into two types: whole cream buttermilk and cheese whey cream buttermilk (WCB). Due to the high casein micelle content of whole cream buttermilk, the removal of casein micelles to improve the purity of MFGM materials is always required. This study investigated the effects of rennet and acid coagulation on the lipid profile of buttermilk rennet-coagulated whey (BRW) and buttermilk acid-coagulated whey (BAW) and compared them with WCB. BRW has significantly higher phospholipids (PLs) and ganglioside contents than BAW and WCB. The abundance of arachidonic acid (ARA)- and eicosapentaenoic acid (EPA)-structured PLs was higher in WCB, while docosahexaenoic acid (DHA)-structured PLs were higher in BRW, indicating that BRW and WCB intake might have a greater effect on improving cardiovascular conditions and neurodevelopment. WCB and BRW had a higher abundance of plasmanyl PL and plasmalogen PL, respectively. Phosphatidylcholine (PC) (28:1), LPE (20:5), and PC (26:0) are characteristic lipids among BRW, BAW, and WCB, and they can be used to distinguish MFGM-enriched whey from different sources.
Subject(s)
Buttermilk , Cheese , Goats , Lipidomics , Whey , Animals , Buttermilk/analysis , Cheese/analysis , Whey/chemistry , Phospholipids/analysis , Phospholipids/chemistry , Glycolipids/chemistry , Milk/chemistry , Lipid Droplets/chemistry , Glycoproteins/chemistry , Glycoproteins/analysis , Lipids/chemistry , Lipids/analysisABSTRACT
The present study aimed to enhance the survivability of the encapsulated biocomposites of Lactiplantibacillus plantarum AB6-25 and Saccharomyces boulardii T8-3C within the gastrointestinal system (GIS) and during storage period. AB6-25 and T8-3C were individually co-encapsulated using either lactobionic acid (LBA) in Na-alginate (ALG)/demineralized whey powder (DWP) or solely potential probiotics in ALG microcapsules. Free probiotic cells were utilized as the control group. Both microcapsules and free cells underwent freeze-drying. The encapsulation and freeze-drying efficiency of core materials were evaluated. The protective effect of encapsulation on the probiotics was examined under simulated GIS conditions and during storage at either 25 °C or 4 °C. Additionally, the microcapsules underwent analysis using fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), and scanning electron microscope (SEM). Encapsulation and freeze-drying processes were carried out efficiently in all groups (88.46 %-99.13 %). SEM revealed that the microcapsules possessed a spherical and homogeneous structure, with sizes ranging from 3 to 10 µm. ALG/DWP and LBA presence in the microcapsule structure was confirmed through FTIR, XRD analysis indicated the formation of a new composite. Over 180 days, all microcapsule groups stored at 4 °C maintained their therapeutic dosage viability. However, after four months, microcapsules stored at 25 °C exhibited a decline in yeast survivability below the therapeutic threshold. Experimental groups demonstrated better viability under simulated GIS conditions compared to the control. These findings suggest the potential use of microencapsulated probiotics as a food supplement and indicate that microcapsule groups containing AB6-25 and T8-3C stored at 4 °C can be preserved for six months.
Subject(s)
Alginates , Capsules , Disaccharides , Probiotics , Saccharomyces boulardii , Whey , Alginates/chemistry , Saccharomyces boulardii/chemistry , Whey/chemistry , Probiotics/chemistry , Disaccharides/chemistry , Freeze Drying , Powders , Lactobacillus plantarum/chemistry , Lactobacillaceae/chemistryABSTRACT
The considerable value of whey is evident from its significant potential applications and contributions to the functional food and nutraceutical market. The by-products were individually obtained during functional chhurpi and novel soy chhurpi cheese production using defined lactic acid bacterial strains of Sikkim Himalaya's traditional chhurpi. Hydrolysis of substrate proteins by starter proteinases resulted in a comparable peptide content in whey and soy whey which was associated with antioxidant and ACE inhibition potential. Peptidome analysis of Lactobacillus delbrueckii WS4 whey and soy whey revealed the presence of several bioactive peptides including the multifunctional peptides PVVVPPFLQPE and YQEPVLGPVRGPFPIIV. In silico analyses predicted the antihypertensive potential of whey and soy whey peptides with strong binding affinity for ACE active sites. QSAR models predicted the highest ACE inhibition potential (IC50) for the ß-casein-derived decapeptide PVRGPFPIIV (0.95 µM) and the Kunitz trypsin inhibitor protein-derived nonapeptide KNKPLVVQF (16.64 µM). Chhurpi whey and soy whey can be explored as a valuable source of diverse and novel bioactive peptides for applications in designer functional foods development.
Subject(s)
Lactobacillus delbrueckii , Peptides , Lactobacillus delbrueckii/metabolism , Peptides/chemistry , Peptides/pharmacology , Angiotensin-Converting Enzyme Inhibitors/chemistry , Angiotensin-Converting Enzyme Inhibitors/pharmacology , Cheese/microbiology , Cheese/analysis , Whey/chemistry , Functional Food , Antioxidants/pharmacology , Antioxidants/chemistry , Whey Proteins/chemistryABSTRACT
Residual lipids (RL) in whey protein isolate (WPI) are detrimental to optimal functional applications (e.g., foaming and low turbidity) and contribute to off-flavor development during powder storage. The objective of this research was to prepare an experimental WPI by removing RL without using the traditional microfiltration process and compare its properties with commercially available WPI made using microfiltration and some other whey powders. We hypothesize that by adjusting the pH of whey to <5.0, we would be close to the isoelectric point of any remaining denatured proteins (DP) and phospholipoproteins (PLP), and therefore reduce electrostatic repulsion between these molecules. Furthermore, demineralization of the acidified whey protein solution by UF combined with diafiltration (DF) should reduce ionic hindrance to aggregation and thereby help with the aggregation of these DP as well as most RL; centrifugation or clarification could be used to remove these materials. Calcium should also be more extensively removed by this approach, which should improve the heat stability of the experimental WPI. Demineralization was achieved on a pilot scale by acidifying liquid (cheese) whey protein concentrate containing 34% protein (WPC-34) to pH 4.5 using HCl, and UF of the whey protein solution along with extensive DF using acidified (pH â¼3.5) reverse osmosis filtered water. Demineralized whey protein solution was adjusted to various combinations of pH (4.1-4.9), conductivities (500-2,000 µS/cm), and protein concentrations (1%-7%) and then centrifuged at 10,000 × g for 10 min. The effective sedimentation (precipitation) of RL in these treatments was estimated by measuring the turbidity of the supernatants. Maximum precipitation was observed at pH 4.5 to 4.7. Reducing conductivity via UF/DF increased the precipitation of RL due to reduced ionic hindrance to aggregation. Maximum sedimentation of RL was observed at protein concentrations ≤3% because of a higher density difference between the precipitate and serum phase. Analysis by sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis confirmed the sedimentation of phospholipoproteins, caseins, and DP upon isoelectric precipitation at pH â¼4.5, while native whey proteins or undenatured whey proteins remained soluble in the supernatant, unaffected by the pretreatment. To scale up the process, 750 L of fluid WPC-34 was acidified and demineralized by UF (volume concentration factor = 1.35) and DF until the permeate solids reached 0.1% (when desired demineralization was achieved), clarified using a pilot-scale desludging clarifier to remove RL, neutralized, ultrafiltered to concentrate the protein, and then spray-dried to produce an experimental WPI (91% protein and 1.8% fat on a dry basis [db]). In another trial, demineralized UF concentrate was clarified by gravity sedimentation and the supernatant was neutralized, ultrafiltered, and spray-dried to produce a second experimental WPI (91% protein and <1% fat db). These experimental WPI powders were compared with several commercially available WPI powders to assess functional properties such as solubility, heat stability, foamability and foam strength, gelation, and sensory attributes over accelerated storage. Experimental WPI had excellent functional properties, had low turbidity, were highly heat stable, and only developed very slight-to-slight off-flavors upon accelerated storage, and their properties were comparable to the WPI manufactured commercially using microfiltration even after accelerated storage.
Subject(s)
Whey Proteins , Whey Proteins/chemistry , Milk Proteins/chemistry , Hot Temperature , Hydrogen-Ion Concentration , Chemical Precipitation , Animals , Whey/chemistryABSTRACT
This study investigated the characteristics of Lactobacillus helveticus-derived whey-calcium chelate (LHWCC) and its effect on the calcium absorption and bone health of rats. Fourier-transform infrared spectroscopy showed that carboxyl oxygen atoms, amino nitrogen atoms, and phosphate ions were the major binding sites with calcium in LHWCC, which has a sustained release effect in simulated in vitro digestion. LHWCC had beneficial effects on serum biochemical parameters, bone biomechanics, and the morphological indexes of the bones of calcium-deficient rats when fed at a dose of 40 mg Ca/kg BW for 7 weeks. In contrast to the inorganic calcium supplement, LHWCC significantly upregulated the gene expression of transient receptor potential cation V5 (TRPV5), TRPV6, PepT1, calcium-binding protein-D9k (Calbindin-D9k), and a calcium pump (plasma membrane Ca-ATPase, PMCA1b), leading to promotion of the calcium absorption rate, whereas Ca3(PO4)2 only upregulated the TRPV6 channel in vivo. These findings illustrate the potential of LHWCC as an organic calcium supplement.
Subject(s)
Bone and Bones , Calcium , Lactobacillus helveticus , Animals , Rats , Calcium/metabolism , Bone and Bones/metabolism , Bone and Bones/drug effects , Male , Rats, Sprague-Dawley , Whey/chemistry , TRPV Cation Channels/metabolism , TRPV Cation Channels/genetics , Calcium, Dietary/pharmacology , Calcium, Dietary/administration & dosage , Dietary Supplements , Calcium Channels/metabolism , Calcium Chelating Agents/pharmacologyABSTRACT
The experiments reported in this research paper aimed to evaluate the physico-chemical and sensory characteristics, microbial quality and antioxidant potential of goat's milk paneer during storage (0-12 d, 4 ± 1°C). The juices from five different citrus fruits were used as coagulant (treatments) to make goat's milk paneer. The pH of all paneer samples decreased during storage whereas titratable acidity increased. Ash (%) fat (%) and protein (%) of paneer increased slightly during storage, whereas sensory perception decreased. The juices from all the citrus fruit varieties showed high contents of total phenolics and total flavonoids which ultimately influenced ferric reducing antioxidant power, total antioxidant capacity and radical scavenging activities. As the contents of different juices were also retained in the paneer matrix, so paneer coagulated with citrus juices also showed encouraging results in terms of total phenolic and flavonoid contents, ferric reducing antioxidant power and radical scavenging activities. Amongst all the paneers, the most promising was that coagulated by kinnow juice. In addition, the whey obtained from paneer coagulated by citrus juices also showed appreciable quantities of total phenolic and flavonoid contents, thereby beneficially influencing ferric reducing antioxidant power andradical scavenging activities. It is concluded that citrus juices improve the sensorial quality and antioxidant potential of goat's milk paneer and its whey.
Subject(s)
Antioxidants , Citrus , Flavonoids , Fruit and Vegetable Juices , Goats , Milk , Phenols , Whey , Animals , Citrus/chemistry , Antioxidants/analysis , Fruit and Vegetable Juices/analysis , Flavonoids/analysis , Milk/chemistry , Whey/chemistry , Phenols/analysis , Food Storage , Food Handling/methodsABSTRACT
SCOPE: In cases where breast milk is unavailable or inadequate, hydrolyzed infant formula is recommended as the primary alternative. The aim of this study is to assess and compare the allergenicity of two partially hydrolyzed whey-based formulas (PHF-Ws) using serum samples from patients with cow's milk allergy (CMA). METHODS AND RESULTS: LC-MS/MS technology is used to investigate the peptide distribution in both samples. The immunoreactivity of two PHF-Ws in 27 serum samples from 50 Chinese infants (02 years) with CMA is analyzed. The results demonstrate that even with a similar a degree of hydrolysis (DH), primary protein sources, peptides with molecular weights <5 kDa, and differences in the number of residual allergenic epitopes in the hydrolyzed peptide segments can lead to varying immune responses. CONCLUSION: The two PHF-Ws have notably high intolerance rates, exceeding 10% among infants with CMA. Therefore, suggesting that PHF-Ws may not be suitable for infants and children with CMA in China.
Subject(s)
Allergens , Infant Formula , Milk Hypersensitivity , Whey Proteins , Humans , Milk Hypersensitivity/immunology , Infant , China , Female , Allergens/immunology , Male , Hydrolysis , Tandem Mass Spectrometry , Whey/chemistry , AnimalsABSTRACT
Perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) are persistent anthropogenic chemicals that are widely distributed in the environment and pose significant risks to human health. Foam fractionation has emerged as a promising method to recover PFOS/PFOA from water. However, PFOS/PFOA concentrations in wastewater are often inadequate to generate stable foams due to their high critical micelle concentrations and the addition of a cosurfactant is necessary. In this study, we developed whey soy protein (WSP) as a green frother and collector derived from soybean meal (SBM), which is an abundant and cost-effective agro-industrial residue. WSP exhibited excellent foaming properties across a wide pH range and demonstrated strong collection capabilities that enhanced the recovery of PFOS/PFOA. The mechanism underlying this collection ability was elucidated through various methods, revealing the involvement of electrostatic attraction, hydrophobic interaction, and hydrogen bonding. Furthermore, we designed a double plate internal to improve the enrichment of PFOS/PFOA by approximately 2.3 times while reducing water recovery. Under suitable conditions (WSP concentration: 300 mg/L, pH: 6.0, air flowrate: 300 mL/min), we achieved high recovery percentages of 94-98% and enrichment ratios of 7.5-12.8 for PFOS/PFOA concentrations ranging from 5 to 20 mg/L. This foam fractionation process holds great promise for the treatment of PFOS/PFOA and other per- and polyfluoroalkyl substances.
Subject(s)
Alkanesulfonic Acids , Fluorocarbons , Water Pollutants, Chemical , Humans , Water , Soybean Proteins , Whey/chemistry , Whey Proteins , Fluorocarbons/analysis , Caprylates/analysis , Alkanesulfonic Acids/analysis , Water Pollutants, Chemical/analysisABSTRACT
Buffalo colostrum is the initial mammary secretion after parturition, consisting of nutritional and bioactive components. In this study, we conducted a proteomic analysis of buffalo colostrum whey to identify bioactive proteins and peptides. A total of 107 differentially expressed proteins (DEPs) were identified in buffalo colostrum whey compared to those in mature milk. Gene Ontology analysis revealed that DEPs were primarily associated with immune response and tissue development. KEGG pathway enrichment suggested that colostrum actively enhances nascent immunity involved in interleukin and interferon signaling pathways. Furthermore, candidate antimicrobial peptides (AMPs) of whey protein hydrolysates from buffalo colostrum were characterized, which exhibits broad-spectrum activity against gram-positive and gram-negative pathogens. Overall, this study improves our understanding of protein variations in buffalo lactation, and contributes to the development of AMPs from buffalo colostrum.