A possible structure of the casein micelle based on high-resolution field-emission scanning electron microscopy

New electron micrographs, produced using the technique of Field Emission Scanning Electron Microscopy, showing the details of the micellar surface, are presented. The images show the micellar surfaces without any coating, and suggest that the surface of the micelle may have a much more complex structure than has previously been demonstrated. Although there appear to be no spherical subunits (submicelles), there is evidence for the organization of the caseins into tubular structures within the micelle. The surface is not smooth, and contains gaps between the substructures. The observations are discussed in terms of published models of micellar structure, where is it suggested how the depiction of the micellar surface can be used to explain certain factors of its reactivity and behaviour.     

A localized surface plasmon resonance based immunosensor for the detection of casein in milk

In this research, a localized surface plasmon resonance (LSPR) immunosensor based on gold-capped nanoparticle substrate for detecting casein, one of the most potent allergens in milk, was developed. The fabrication of the gold-capped nanoparticle substrate involved a surface-modified silica nanoparticle layer (core) on the slide glass substrate between bottom and top gold layers (shell). The absorbance peak of the gold-capped nanoparticle substrate was observed at ∼520 nm. In addition, the atomic force microscopy (AFM) images demonstrated that the nanoparticles formed a monolayer on the slide glass. After immobilizing anti-casein antibody on the surface, our device, casein immunosensor, could be applied easily for the detection of casein in the raw milk sample without a difficult pretreatment. Under the optimum conditions, the detection limit of the casein immunosensor was determined as 10 ng/mL. Our device brings several advantages to the existing LSPR-based biosensors with its easy fabrication, simple handling, low-cost, and high sensitivity.     

UHT milk contains multiple forms of αS1-casein that undergo degradative changes during storage

Milk proteins are susceptible to chemical changes during processing and storage. We used proteomic tools to analyse bovine α_S1-casein in UHT milk. 2-D gels of freshly processed milk α_S1-casein was presented as five or more spots due to genetic polymorphism and variable phosphorylation. MS analysis after phosphopeptide enrichment allowed discrimination between phosphorylation states and genetic variants. We identified a new alternatively-spliced isoform with a deletion of exon 17, producing a new C-terminal sequence, K¹⁶⁴SQVNSEGLHSYGL¹⁷⁷, with a novel phosphorylation site at S¹⁷⁴. Storage of UHT milk at elevated temperatures produced additional, more acidic α_S1-casein spots on the gels and decreased the resolution of minor forms. MS analysis indicated that non-enzymatic deamidation and loss of the N-terminal dipeptide were the major contributors to the changing spot pattern. These results highlight the important role of storage temperature in the stability of milk proteins and the utility of proteomic techniques for analysis of proteins in food.     

The production of volatile compounds in model casein systems with varying fat levels as affected by low-frequency ultrasound

The effects of low-frequency ultrasound on the production of volatile compounds in model casein protein systems containing various fat concentrations of 2%, 4% and 6% (w/w) were investigated. Ultrasound application was performed at 20 kHz for up to 10 min which corresponded to energy densities ranging from 9.54 to 190.8 J mL⁻¹. Similar volatile compounds were detected both in pure fat and mixtures of casein and fat (CF) systems. These volatiles belonged to the groups of aldehydes, ketones, esters, alcohols and hydrocarbons, which were the products of oxidation of lipids or protein degradation due to acoustic cavitation. The amount of fat in the casein systems had minor effects on the production of volatiles, whereas the production of volatile compounds was significantly affected by the ultrasound treatment. Short sonication times <5 min generated similar volatile profiles to the untreated samples. In contrast, prolonged sonication for 5 and 10 min considerably increased the production of volatile compounds and the amounts of fatty acids. Thus, the application of low–frequency ultrasound for short periods should be considered to minimise the production of volatile compounds which can ultimately affect the taste.     

Polyethylene/Polycaprolactone Nanocomposite Films for Food Packaging Modified with Magnetite and Casein: Oxygen Barrier,Mechanical, and Thermal Properties

Polyethylene (PE) was modified and prepared as double-layer polyethylene/polycaprolactone (PE/PCL) film. Magnetite and casein were added to the PCL-coating film to improve barrier properties and prevent destruction of basic structure of primary polymer PE. Significant improvements were observed with regards to mechanical (tensile strength, elongation at break) and thermal properties, while barrier (O₂ permeability) properties were slightly improved. Overall migration values into acetic acid were lower (from 1 to 4.6 mg/dm²) than the upper limit set by the legislation. Specific migration of iron in PE/PCL-Fe samples is also below (µg/L) specific migration limit value set by the legislation (mg/kg).     

Foaming and surface properties of casein glycomacropeptide–gelatin mixtures as affected by their interactions in the aqueous phase

Gelatin is widely used in food industry for the stabilization of foam products. On the other hand, casein glycomacropeptide (CMP) is a bioactive peptide with high surface activity. The aim of this work was to study the interfacial and foaming properties of CMP–gelatin mixed systems at pH 6.5 and 3.5 and evaluate the relation of these properties with the interactions in the aqueous phase. The CMP:gelatin ratio in mixed systems was 0:100 (pure gelatin), 25:75, 50:50, 75:25 and 100:0 (pure CMP). Viscosity, particle size, ζ-potential, interfacial properties and foaming properties were determined. At both pH, gelatin solutions showed the highest viscosity, CMP the lowest and the mixed systems presented behaviour more similar than CMP. Particle size and ζ-potential determinations evidenced the formation of complexes between CMP and gelatin at both pH. CMP was more surface active than gelatin and dominated the rate of diffusion of the mixed systems to the air–water interface. A synergistic effect was observed on foamability and foam stability in mixed systems that could be explained by the formation of a complex between CMP and gelatin with outstanding capacity for foams formation and stabilization.     

Stabilisation mechanism of various inulins and hydrocolloids: Milk–sour cherry juice mixture

Milk–fruit juice mixtures, such as the mainly acidic nutraceutical soft drinks, usually suffer from phase separation due to aggregation of caseins at low pH. In this study, short‐chain inulin (SCI), native inulin (NI), long‐chain inulin (LCI) and a combination of long‐ and short‐chain inulins (LCI:SCI) (MIX) in different ratios (20:80, 50:50 and 80:20) were added (up to 10% w/v) to a milk–sour cherry juice mixture and their stabilisation mechanisms investigated using rheological, microstructural and zeta potential observations. In addition, gum tragacanth (GT) and Persian gum (PG) as adsorbing and guar gum (GG) as nonadsorbing hydrocolloids were combined with inulin to enhance their stabilising properties. Finally, sensory analyses were carried out on the stabilised samples. According to our findings, LCI fully stabilised the mixture (8% w/v), while LCI: SCI and NI only reduced phase separation at very high concentrations, and SCI had no significant effect on the stabilisation. Moreover, no inulin aggregates and rheological changes were observed with SCI. However, LCI, LCI: SCI and NI formed inulin aggregates and the mixtures became even more viscous and thixotropic (LCI > LCI: SCL > NI). Based on these observations, it can be concluded that chain length and concentration are two important factors that affect the functionality of inulin. On the other hand, the combination of inulin with GT and PG did not have any pertinent effect on the stabilisation. However, the mixture of inulin and GG could stabilise the mixtures at certain ratios and concentrations. Furthermore, in mixtures containing GG and SCI, GG played the main role in the stabilisation by increasing the viscosity and forming gel network.     

Changes in the physico-chemical properties of casein micelles during ultrafiltration combined with diafiltration

The objective of this work was to determine if after membrane processing, the physical properties of casein micelles change. Milk was concentrated by ultrafiltration, and also subjected to various levels of diafiltration, by addition of water to the retentate. After the membrane concentration process, the retentates were diluted back to their original concentration, to study their physico-chemical properties. For better comparison, all the samples were dialyzed against the original milk, to obtain similar serum compositions. For the first time, the effect of different levels of diafiltration was studied. Diafiltration induced losses of colloidal calcium phosphate and caused changes in the turbidity parameter (1/l*) measured by light scattering, as well as in the ultrasonic properties (velocity and attenuation) of the casein micelles. When tested in a similar serum environment, the reconstituted micelles after diafiltration showed a lower susceptibility to aggregation and the rennet induced gels had a lower storage modulus than those formed with the original milk, at the same protein concentration. This work brings for the first time evidence of the differences in the physical properties of the casein micelles as a function of membrane processing history.     

Effect of fluoride varnish with added casein phosphopeptide-amorphous calcium phosphate on bond strength to enamel

To investigate the effect of fluoride varnish with added casein phosphopeptide-amorphous calcium phosphate on the shear bond strength (SBS) of two adhesive systems to enamel. Specimens obtained from permanent teeth were randomly distributed among four groups for enamel pretreatment [Control (no treatment, CNT), Duraphat varnish (DV), Clinpro White varnish (CWV), MI Varnish (MIV)], and each group was further divided into two subgroups according to adhesive [Etch&rinse (Adper Single Bond, ASB), self-etch (Clearfil SE Bond, CSE)]. Specimens were stored in distilled water at 37 °C for 24 h. Cylindrical composite specimens (2.3 mm in diameter, 3.0 mm in height) were then bonded to the enamel surfaces. SBS tests were performed and data were analyzed with two-way ANOVA and Tukey’s tests. For both CSE and ASB, SBS values of the CNT groups were significantly higher than those of all the enamel pretreatment groups (p < 0.05). Among the enamel pretreatment groups, SBS values with both adhesive systems were lowest in the MIV groups, followed by CWV and DV groups. In conclusion, pretreatment of enamel surfaces with fluoride-containing varnishes reduced bonding performance of adhesive systems to enamel. MIV appeared to cause greater enamel surface alterations and precipitation, which interfered with adhesive bonding mechanisms.