Cow Whey Research Articles

Algerian Yeast Strains: Isolation, Identification and Production of Single Cell Protein from Whey with Strain Candida kefyr

In this study, few samples than soft and baked cheese, yogurt, cow's milk, pasteurized milk and cheese whey were collected from dairy industries in the city of Boumerdes. The samples were cultured on yeast extract glucose chloramphenicol agar (YGCA) media. Twenty yeast strains were isolated from the culture. Tenof theses strains were identified by their morphological and physiological properties. Betagalactosidase activity in the yeast strains showed that a strain Candida kefyr designated as LP1 had highest enzyme activity (up to 5000 EU/ml). To investigate the effectiveness of batch submerged fermentation of the yeast C. kefyr in cheese whey, we found that this strain have highest level of single cell protein (SCP), production (up to 19 g/l dry mass cell). The isolated yeast strain was examined for his ability in reduction of the chemical oxygen demand (COD). So, the results demonstrated a high level of reduction.

The effect of storage on nutritional, textural and sensory characteristics of creamy ricotta made from whey as well as cow's milk and goat's milk

AbstractThe aim of this study was to develop a creamy ricotta using a mixture of goat and cow whey as the main ingredients, with the addition of whole goat and cow milk. The nutritional composition, texture and sensory characteristics of the ricotta cheese were evaluated over 14 days of refrigerated storage. There was a decrease in protein and ash content and pH changes during the assessed storage periods. The instrumental texture profile indicated that the creamy ricotta was easily deformable, with minimal inelasticity and a cohesive, soft and delicate texture. The medium‐ and long‐chain fatty acid content was higher than the short‐chain fatty acid content. The flavour and aroma of creamy ricotta were described by the panel as having a soft texture and wet appearance. The use of whey from goat and cow cheese in making creamy ricotta is feasible and does not cause negative impact in the final product.

PRODUCTION OF LACTIC ACID FROM SWEET MEAT INDUSTRY WASTE BY LACTOBACILLUS DELBRUKI

A large amount of whey is discharged from sweet meat industry, which is responsible for environmental pollution and a large amount of whey protein and milk sugar are also wasted. This whey may be utilized for valuable lactic acid production. Lactobacillus delbruki was used for lactic acid production from cow-milk whey. Lactic acid production was 12.22 gm/L at pH: 6.8, temperature: 42 0 C, Inculum volume: 4% , fermentation time: 24hr, medium volume: 125 mL in 250 mL Erlenmeyer flask, medium composition, whey supplemented with peptone : 1.5%, glucose: 2.0% and ammonium chloride: 1.0%.

Manufacture of bioactive peptide-rich concentrates from Whey: Characterization of pilot process

This work was focused on the manufacture, at pilot scale, of cow whey protein and peptide concentrates, using selective filtration techniques—associated with hydrolysis brought about by proteolytic enzymes from Cynara cardunculus aqueous extracts, using as (optimal) conditions an enzyme/substrate ratio of 1.6%v/v, a pH of 5.2, a temperature of 55°C and an incubation time of 7h. The profiles of proteins and peptides were assessed by liquid chromatography and electrophoresis; ca. 87% of α-lactalbumin was hydrolyzed, but essentially no degradation of β-lactoglobulin (β-Lg) was observed. A bioactive peptide concentrate, its fraction below 3kDa and a β-Lg-rich fraction were obtained as final products, containing ca. 73, 43 and 91%w/w protein (on a total solid mass basis). All these fractions were low in lactose and salt, and their microbial loads were reduced. Said fractions are high added-value products, so they can be used as nutritional and functional ingredients—thus yielding an economically viable alternative for upgrade of whey.

Innovation in the development of fermented dairy drink with cow's milk and goat cheese whey

The fermented dairy drink is an important form of utilization of whey because of its high acceptance. Passion fruit and bee honey are regional products with features that can be exploited in the development of the drink, as acidity and its functional potential, respectively. The objective of this research is to study the various relationships of mixing milk and whey in the preparation of a dairy drink flavored with passion fruit and sweetened with honey of Africanized bees, as well as to characterize this drink according to its physicochemical, microbiological and sensory aspects and to evaluate its stability and purchase intent. There was no difference in the sensory aspects among adults about the content of pulp and the goat whey that were tested. Regarding the acceptance test with children, 88% of the participants said they liked the drink. Regarding the stability of the product, there was a decrease in the pH from 4.6 to 3.8 during the first seven days, but it remained stable during the storage. In the five evaluated times, there was no significant difference among the sensory aspects, except because of one flavor that decreased until the 14th day of storage, and after that a slight growth was observed, returning to the acceptance level found on the 7th day of storage. The passion fruit flavored drink showed considerable values for purchase intent, suggesting to be a food with market potential.

Proteomic characterization by 2-DE in bovine serum and whey from healthy and mastitis affected farm animals

Acute phase proteins (APP) have been identified in whey and sera from healthy and mastitis cows through the proteomic analysis using two-dimensional electrophoresis (2-DE) coupled with Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS). Although normal and mastitis serum samples show relatively similar protein composition, marked differences in expression levels and patterns can be observed. Conversely, normal and mastitis whey showed a very different composition, likely due to extravasation of blood proteins to the mammary gland. Different isoforms from the most abundant protein in milk, casein, were detected in both normal and mastitis whey. Other proteins, such as lactotransferrin, were only detected in the inflamed animal samples. Immunoglobulins showed different patterns but not increased levels in the inflamed whey. Also, many cellular proteins in mastitis cow's whey, that were absent from healthy cow's milk. They are responsible for the great change in composition between normal and mastitis whey, especially those which exert a biological function related to immune defense. Data collected in this work are of interest for gaining information about physiological changes in protein patterns in different fluids and, the correspondent modifications as result of an acute phase process in farm. This article is part of a Special Issue entitled: Proteomics: The clinical link.

Studying the distribution of Selenium in buffalo and cow's milk whey

In this study, size exclusion chromatography (SEC) coupled with an online ultraviolet detection (UV) and graphite furnace atomic absorption spectrometry (GF AAS) and matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS) were used to study the association of selenium with macromolecules and compounds present in buffalo and cow's milk whey. SEC-UV profiles obtained from buffalo and cow's milk whey samples indicate the presence of high molecular weight (HMW) and low molecular weight (LMW) species in both samples studied. The combined information obtained with SEC-UV, GF AAS and MALDI-TOF-MS for fractions < 10 kDa confirmed the association of selenium with LMW species

Evaluation of genetic polymorphism among Lactobacillus rhamnosus non-starter Parmigiano Reggiano cheese strains

Parmigiano Reggiano (PR) is an Italian cooked, long-ripened cheese made with unheated cow's milk and natural whey starter. The microflora is involved in the manufacturing of this cheese, arising from the natural whey starter, the raw milk and the environment. Molecular studies have shown that mesophilic non-starter lactic acid bacteria (NSLAB) are the dominant microflora present during the ripening of PR. In this study, a characterisation of Lactobacillus rhamnosus isolated from a single PR manufacturing and ripening process is reported, using a combination of genotypic fingerprinting techniques (RAPD-PCR and REP-PCR). The intraspecies heterogeneity evidenced for 66 strains is correlated to their abilities to adapt to specific environmental and technological conditions. The detection of biotypes that correlate with specific moments in cheese ripening or differential development throughout this process suggests that these strains may have specific roles closely linked to their peculiar technological properties.

The Bovine Lactophorin C-Terminal Fragment and PAS6/7 Were Both Potent in the Inhibition of Human Rotavirus Replication in Cultured Epithelial Cells and the Prevention of Experimental Gastroenteritis

Rotaviruses are the leading cause of severe dehydrating diarrhea in children worldwide. We have found that high-M(r) glycoprotein fraction (F1) from cow's milk whey has potent inhibitory activity against human rotavirus (HRV) in cell culture. The present study was undertaken to identify and characterize the components responsible for this inhibitory activity. F1 was initially heated at 95 degrees C for 30 min, rendering milk antibodies inert, subjected to ammonium sulfate fractionation, and then resolved by two-dimensional polyacrylamide gel electrophoresis. After electroelution, we found that a heat-stable milk protein lactophorin C-terminal fragment (LP16) and bovine milk fat globule membrane protein PAS6/7 strongly inhibited the replication of HRV MO strains in MA104 cells. Furthermore, we found that prophylactic oral administration of F1 once before inoculation of the HRV MO strain obviously prevented the development of diarrhea in vivo. These non-immunoglobulin components are a promising candidate for a prophylactic food additive against HRV infection.

Contribution of IgE and immunoglobulin free light chain in the allergic reaction to cow's milk proteins

Cow's milk allergy (CMA) affects 2.5% of young infants. In previous murine studies it was observed that allergic sensitization to the major cow's milk allergens casein and whey led, respectively, to IgE-independent and IgE-dependent clinical responses. In this study the involvement of immunoglobulin free light chains (Ig-fLCs) in the hypersensitivity response to cow's milk proteins was explored in mice, and Ig-fLC serum levels were determined in children affected by CMA or atopic dermatitis (AD). Mice were orally sham, casein, or whey sensitized. Acute allergen-specific skin responses were determined, and serum immunoglobulin and Ig-fLC concentrations were measured. Ig-fLC dependency was validated by using the Ig-fLC blocker F991 in actively and passively sensitized mice. Ig-fLC serum concentrations were measured in a cohort of infants with CMA and infants with AD. After sensitization, no specific IgE was detectable in sera of casein-sensitized mice, whereas specific IgE levels were enhanced in whey-sensitized mice. Instead, Ig-fLC levels were increased in sera from casein-sensitized mice. Furthermore, blocking Ig-fLCs strongly diminished the allergic skin responses not only in casein-sensitized mice but also in mice transferred with splenocyte supernatants of casein-sensitized mice. In both patients with CMA and patients with AD, serum Ig-fLC concentrations were significantly enhanced. This study indicates that sensitization with cow's milk proteins can lead to both IgE-dependent and Ig-fLC-dependent allergic hypersensitivity responses. Also, in children affected with CMA or AD, serum Ig-fLC concentrations were increased, implying the relevance of Ig-fLC measurements in the diagnoses of human allergic disease.

The Multiplicity ofN-Glycan Structures of Bovine Milk 18 kDa Lactophorin (Milk GlyCAM-1)

Lactophorin is a heat-stable phosphoglycoprotein, also known as milk glycosylation-dependent cell adhesion molecule 1 (GlyCAM-1). Bovine 18 kDa lactophorin was purified by heparin affinity chromatography from cow's milk whey. Its N-glycans were obtained by proteomic techniques, including two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), followed by in-gel digestion with peptide-N(4)-(N-acetyl-beta-glucosaminyl)-asparagine amidase (PNGase F). The released N-glycans were derivatized with 2-aminopryridine (PA) and analyzed by matrix-assisted laser desorption ionization quadruple ion trap time of flight mass spectrometry (MALDI-QIT-TOF MS). Among the MS analyzed peaks, 15 peaks were found to be N-glycan molecules as detected by MS(2) analysis. These glycans consisted of mono-sialylated bi-, tri-, and tetra-antennary complex-type N-glycans carrying Gal-GlcNAc (LacNAc) or GalNAc-GlcNAc (LacdiNAc) with and without core-fucose.

Protein composition of yak milk

The protein composition of 24 individual Maiwa yak milk samples that were collected in the Sichuan province of China was determined by reversed-phase high-performance liquid chromatography. Yak milk differed from cow milk by its high concentration of total proteins (46.2–58.4 g·L−1), total casein (40.2 g·L−1 on average) and the proportion of individual caseins. The high content of β-casein (more than 45%) and consequently the lower proportion of αs-casein (about 40%) together with a small increase in κ-casein (15%) appear to make this milk more favorable for infant nutrition as used by the Tibetan nomads. Also, the whey proteins, β-lactoglobulin and serum albumin, showed similarities in centesimal composition with the homologous cow whey proteins. However, this initial study on yak milk proteins has highlighted the lack of knowledge in the literature and underlines that much research is required to optimize the utilization of all components of this milk for people living in extreme climatic environments.

In-Depth Exploration of Cow’s Whey Proteome via Combinatorial Peptide Ligand Libraries

The use of combinatorial peptide ligand libraries, containing hexapeptides terminating with a primary amine, or modified with a terminal carboxyl group, allowed discovering and identifying a large number of previously unreported proteins in cow's whey. Whereas comprehensive whey protein lists progressively increased in the last 6 years from 17 unique gene products to more than 100, our findings have considerably expanded this list to a total of 149 unique protein species, of which 100 were not described in previous proteomics studies. As an additional interesting result, a polymorphic alkaline protein was observed with a strong positive signal when blotted from an isoelectric focusing separation in gel and tested with sera of allergic patients. This polymorphic protein, found only after treatment with the peptide library, was identified as an immunoglobulin (Ig), a minor allergen that had been largely amplified. The list of cow's whey components here reported is by far the most comprehensive at present and could serve as a starting point for the functional characterization of low-abundance proteins possibly having novel pharmaceutical, diagnostic, and biomedical applications.

Influence of Alcalase and transglutaminase on immunoreactivity of cow milk whey proteins

The aim of the research was to determine the changes in the immunoreactivity of whey protein concentrate (WPC) modified by two enzymes: proteinase, Alcalase 2.4L FG (Novo Nordisk), and cross-linked transglutaminase (EC 2.3.2.13, Activa&lt;sup&gt;TM&lt;/sup&gt; P, m-TG, Ajinomoto). The new products were characterised by 2D electrophoresis, immunoblotting, and ELISA methods. The WPC hydrolysate obtained with Alcalase contained proteins and peptides characterised mostly by low molecular weight peptides (MW &lt; 14.4 kDa) in the pH range of 3–10. Immunoblotting showed strong immunoreactive properties of the hydrolysate with &amp;alpha;-la and &amp;beta;-lg polyclonal rabbit antibodies. The 2D electrophoretic patterns of WPC and its modified product obtained with m-TG did no differ significantly. However, the immunoblot analysis demonstrated that WPC showed a stronger reactivity towards IgE of allergic patients as compared to WPC with m-TG. ELISA methods showed that two-step hydrolysis with Alcalase followed by m-TG significantly reduced the immunoreactive properties of whey proteins. No cross reactions were observed with α-la and only about 0.6% cross-reactivity with &amp;beta;-lg.

Dietary Whey Protein Hydrolysate Suppresses Development of Atopic Dermatitis-like Skin Lesions Induced by Mite Antigen in NC/Nga Mice

Oral administration of enzymatic hydrolysate of cow's milk whey protein (WPH) has been reported to produce an anti-inflammatory effect. Since inflammation plays a role in dermatitis of allergic disease, we examined the influence of WPH on the development of atopic dermatitis (AD)-like skin lesions, induced in NC/Nga mice by the mite antigen Dermatophagoides pteronyssinus (Dp). AD-like skin lesions were induced on the pinnae and backs of NC/Nga mice by daily application of Dp for 4 weeks. Mice were fed cow's milk casein (control), WPH or casein protein hydrolysate (CPH) diets for 2 weeks prior to Dp application. Clinical skin conditions were evaluated periodically by a clinical severity score, total serum IgE and soluble E-selectin levels were measured by enzyme linked immunosorbent assay (ELISA). WPH-fed mice showed significantly less AD-like skin lesions than those fed casein diets at 2 and 4 weeks after Dp application. In contrast, CPH-fed mice had manifestations in a similar manner as casein-fed mice did, and did not show an inhibitory effect. Serum soluble E-selectin levels, known as a marker of disease activity in AD patients, were significantly lower in the WPH diet group. Our results suggest that in addition to its hypoallergenicity an anti-inflammatory function, dietary WPH might be useful for reducing the severity of AD-like skin lesions.

Concentration of alpha-lactalbumin from cow milk whey through expanded bed adsorption using a hydrophobic resin

The design of novel applications for the use of new high-density adsorbents in the direct recovery of proteins requires analysis of the fluid dynamics and mass transfer characteristics of the column used with the solid-liquid system. In this article we describe the operating parameters related to the use of a high-density, hydrophobic resin (Streamline® Phenyl) for the recovery of alpha-lactalbumin from cow milk whey in an expanded-bed adsorption mode of operation. The adsorption isotherm, kinetic curves and chromatographic curves for adsorption and elution were determined for alpha-lactalbumin recovered from whole whey. The experiments were used to confirm the theoretical model that involves the combination of resistance to the mass transfer in the liquid phase and diffusivity in the adsorbent resin pores. The model solution was obtained through the orthogonal collocation method. A nonlinear multivariable optimization method was used to fit the model to experimental results. A purity of 79% was obtained with the adsorption/elution protocol prior to any polishing procedure.

Possible involvement of selenium inStaphylococcus aureusinhibition in cow's whey

Previous investigations have shown that selenium supplementation inhibits growth of mastitis pathogens in cow's milk. The present study was performed to clarify the role of selenium in defence mechanisms of mammary gland. We have examined the effects of selenium supplementation on Staphylococcus aureus growth inhibiting activity in whey. Six selenium-supplemented and six non-supplemented Estonian dairy cows were used for this study. Selenium-supplemented cows received 4 mg organic selenium in the form of selenium yeast in their daily diet for 8-week period. Cows from non-supplemented group received the same amount of yeast free of selenium in their diet. All cows had initially low blood glutathione peroxidase (GPx) activity (<or=1.02 microkat/g Hb). During the 8-week experiment these cows had low somatic cell count (<300,000 cells/ml) and negative bacteriological findings in their quarter milk samples. Bacterial growth inhibitory activity in specific whey fractions was clearly evident in 3 out of 6 cows supplemented with selenium. The inhibitory effect was more profound if the cow's blood GPx activity exceeded 4 microkat/g Hb, whereas in non-supplemented cows the inhibition of S. aureus growth was not observed. This study confirms the presence of selenium-dependent antibacterial activity in cow's milk.

Immunoreactive properties of peptide fractions of cow whey milk proteins after enzymatic hydrolysis

SummaryCommercial whey protein concentrate (WPC) was hydrolysed with either Alcalase 2.4 FG (Novo Nordisk), or papain (Sigma) (in one‐step process) or with two enzymes (in two‐step process) to determine the changes in the immunoreactivity of α‐lactalbumin and β‐lactoglobulin. Enzymatic hydrolysis of WPC was performed by pH‐stat method. Hydrolysates were analysed using sodium dodecyl sulphate‐polyacrylamide gel electrophoresis, immunoblotting and size‐exclusion chromatography (SE‐HPLC). Immunoreactive properties of peptide fractions separated from the hydrolysates by fast protein liquid chromatography (FPLC) were determined using dot‐immunobinding and enzyme‐linked immunosorbent assay (ELISA) methods. Finally the sensory analysis was used to confirm organoleptic changes resulting from the application of different enzymes. The ‘two‐step’ process was observed to be the most effective however allergenic epitopes were still present, as it was found by ELISA with anti‐α‐la and anti‐β‐lg antibodies. The addition of papain as the second enzyme in the hydrolysis process contributed to the improvement of the sensory properties of WPC hydrolysate as compared with the Alcalase hydrolysate. Alcalase‐papain partially hydrolysated WPC can be found a promising base for production of the tolerogenic formula.

Safety of milk and milk derivatives in relation to BSE: the lactoferrin example.

Bovine Spongiform Encephalopathy (BSE) belongs to Transmissible Spongiform Encephalopathies (TSEs) or Prion diseases. BSE is a feed borne infection of cattle. Epidemiological and laboratory data suggest that the BSE infectious agent is responsible for the variant form of Creutzfeldt-Jakob Disease (vCJD) and that the oral route is the most plausible way of infection. Therefore there is concern that the BSE agent can be transmitted to humans by biological materials (i.e. meat products, blood, milk) from susceptible BSE animal species (mostly cows but possibly, sheep and goats). Lactoferrin (LF) can be produced by purification from large volumes of cow's milk or whey. Therefore, a potential BSE risk for milk and milk products needs to be evaluated by risk assessment. The Committee for proprietary Medicinal Products--CPMP of the European Commission and the WHO have categorized risk tissues from TSE susceptible ruminant species in different classes in relation to the BSE risk for medicinal products. Milk, colostrum, and tissues of the mammary gland have been classified in the category of no detectable infectivity. A secondary contamination of milk can be virtually excluded (i.e. milk is taken from living animals). In the light of current scientific knowledge and irrespective of the geographical origin, milk and milk derivatives (e.g. lactoferrin, lactose) are unlikely to present any risk of TSE contamination provided that milk is sourced from healthy animals in the same conditions as milk collected from human consumption. So the risk of milk and milk derivatives in relation to BSE is negligible.

Effect of temperature on immunoreactive properties of cow milk whey protein. I. a-lactalbumin

The effect of heat induced changes on the immunoreactivity of a-lactalbumin (a-la) in whey was evaluated. Whey and acidified a-la solutions were heated for various times at temperatures between 60 and 100 °C. After heating, the samples were clarified and the protein content (by Bradford's method), the quantity of a-la (by FPLC method) and the immune response to an anti a-la probe were determined. The samples heated at 80 °C were characterized by the highest a-la immunoreactivity properties. Raising temperature up to 90 °C and 100 °C caused a marked decrease in a-la immunoreactivity. As was found by FPLC method in the samples heated at 60 and 65 °C, a-la content changes were not statistically significant. A gradual decrease in the content of a-la was observed with increased temperature and duration of heating.