Effects of lipids on the water sorption, glass transition and structural strength of carbohydrate-protein systems

peer-reviewedEncapsulant systems are gaining wide practical interest due to their functional and nutritional properties. This paper was focusing on understanding structural relaxations in that systems near glass transition temperature. Freeze-dried trehalose-whey protein isolate-sunflower oil systems with various ratios of the last were used as a carbohydrate-protein-lipid food model. The Guggenheim-Anderson-de Boer (GAB) water sorption relationship was used as a tool to model water sorption isotherms. The glass transition temperature was obtained by differential scanning calorimetry (DSC). Structural α-relaxation temperatures were measured by dynamical mechanical analyses (DMA), dielectric analysis (DEA) and combined to cover a broad range for strength assessment. The microstructure was characterized by optical light microscopy, confocal laser scanning microscopy and scanning electron microscopy. The C1 and C2 constants for Williams-Landel-Ferry (WLF) equation and structural strength parameter were calculated for each system. The effect of sunflower oil and water contents on strength of carbohydrate-protein system was analyzed. Strength shows decreasing with increasing of lipid concentration in the mixtures and more complex dependence on the water content in a system.This investigation was supported by the Food Institutional Research Measure (FIRM) project “Formulation and Design for Food Structure and Stability” funded by the Department of Agriculture, Food and Marine (11-F-001), coordinated by prof. Y.H. Roos, UCC, Ireland and by the Food Institutional Research Measure (FIRM) project “Developing the next generation of high protein spray dried dairy powders with enhanced hydration properties” (15-F-679) funded by the Department of Agriculture, Food and Marine, coordinated by Dr. Mark Auty, Teagasc Food Research Centre, Moorepark, Co. Cork, Ireland

Cytotoxic Complexes of Sodium Oleate with β-Lactoglobulin

pre-printA complex of α-lactalbumin and oleic acid has previously been shown to induce apoptosis in cancer cells in a number of in vitro and in vivo trials. This complex is called HAMLET or BAMLET, depending on the origin of α-la (human/bovine alpha-lactalbumin made lethal to tumour cells). In the current study, it was shown that bovine β-lactoglobulin (β-lg), upon binding sodium oleate (NaOle), the salt of oleic acid, also acquires cytotoxicity towards tumour cells (human monocytic cells U937), analogously to HAMLET/BAMLET complexes. The properties of the complex were characterized using FIR spectroscopy, HPLC and SDS-PAGE. It was shown that the level of covalent oligomerization (dimers and trimers) of β-lg increased with increasing the molar ratio of sodium oleate NaOle:β-lg in the preparation procedure. At the same time, increasing the molar ratio of NaOle:β-lg increased the cytotoxicity of the complex. The increase in cytotoxicity appeared to be dependent on the amount of bound NaOle in the complex, but not on the content of multimeric forms of β-lg. The NaOle/β-lg complex also showed similarity with BAMLET in penetrating the cell membrane and co-localizing with the cell nucleus. Furthermore, DNA fragmentation studies suggested that tumour cells (U937) treated with the complex died by apoptosis, as in the case of BAMLET, and healthy cells appeared to be less affected by treatment, as shown with model rat adrenal pheochromocytoma cells PC12. In conclusion, β-lg and NaOle can form complexes with apoptosis-inducing qualities comparable to those of BAMLET.The work was funded by the Irish Dairy Research Trust and The Department of Agriculture (Food Institutional Research Measure – FIRM project 08RDTMFRC650) under the National Development Plan 2007-2013. K. Lišková was funded under the TeagascWalsh Fellowship Scheme

The Effects of Processing and Ripening on the Quality of Pizza Cheese

End of Project ReportThe main aims of this project were to quantify the changes in fuctionality during maturation of cheese and to develop an understanding of the factors which mediate the development of functionality. The approach to achieving these objectives involved the establishment of a suitable pilot plant production procedure for low moisture Mozzarella, developing and/or adapting existing methods for objective evaluation of the functional properties of pizza cheeses, and evaluating the effects of ripening and variations in cheesemaking conditions (e.g. pH at stretching) on the composition, yield and functionality of low moisture Mozzarella cheese. The main conclusions were as follows: The technology for developing low moisture Mozzarella cheeses, with different compositions and functionalities, via alteration of cheesemaking parameters, has been developed. A database has been established on the storage-related changes that occur in texture, proteolysis and functionality of low moisture Mozzarella cheeses of different compositions. In addition an extensive database on the compositional, biochemical, microstructural, rheological and/or functional properties of different commerical cheeses - low moisture Mozzarella, Cheddar and analogue pizza cheese, has been compiled. The functionality of low moisture Mozzarella changes markedly on storage/ripening at 4ºC. Initially, during the first 5-10 days of storage, the functionality of the baked cheese is poor but then improves on further storage as reflected by reductions in melt time and apparent viscosity (chewiness) and increases in stretchability and flowability. The changes in functionality are mediated by storage-related increases in pH, proteolysis, protein-bound water and free oil in the cheese. On prolonged storage (e.g. > 60 d at 4ºC), the cheese functionality becomes impaired as the shredded cheese develops an increased susceptibility to clumping/balling which makes it difficult to dispense the cheese onto the pizza pie and achieve a uniform surface distribution. Moreover, the baked cheese tends to exude excess free oil and loses its desired level of chewiness attaining a 'soupy' consistency. Novel methods were developed/adapted to objectively quantify functionality in the raw (susceptibility of shredded cheese to clump) and cooked (stretchability, chewiness, viscoelasticity) cheeses.Department of Agriculture, Food and the Marin

Effects of depleting ionic strength on 31P nuclear magnetic resonance spectra of micellar casein during membrane separation and diafiltration of skim milk

peer-reviewedMembrane separation processes used in the concentration and isolation of micellar casein-based milk proteins from skim milk rely on extensive permeation of its soluble serum constituents, especially lactose and minerals. Whereas extensive literature exists on how these processes influence the gross composition of milk proteins, we have little understanding of the effects of such ionic depletion on the core structural unit of micellar casein [i.e., the casein phosphate nanocluster (CPN)]. The 31P nuclear magnetic resonance (NMR) is an analytical technique that is capable of identifying soluble and organic forms of phosphate in milk. Thus, our objective was to investigate changes to the 31P NMR spectra of skim milk during microfiltration (MF) and diafiltration (DF) by tracking movements in different species of phosphate. In particular, we examined the peak at 1.11 ppm corresponding to inorganic phosphate in the serum, as well as the low-intensity broad signal between 1.5 and 3.0 ppm attributed to casein-associated phosphate in the retentate. The MF concentration and DF using water caused a shift in the relevant 31P NMR peak that could be minimized if orthophosphate was added to the DF water. However, this did not resolve the simultaneous change in retentate pH and increased solubilization of micellar casein protein. The addition of calcium in combination with orthophosphate prevented micellar casein solubilization and simultaneously contributed to preservation of the CPN structure, except for overcorrection of retentate pH in the acidic direction. A more complex DF solution, involving a combination of phosphate, calcium, and citrate, succeeded in both CPN and micellar casein structure preservation while maintaining retentate pH in the region of the original milk pH. The combination of 31P NMR as an analytical technique and experimental probe during MF/DF processes provided useful insights into changes occurring to CPN while retaining the micellar state of casein

Covalent labelling of β-casein and its effect on the microstructure and physico-chemical properties of emulsions stabilized by β-casein and whey protein isolate

peer-reviewedThe objective of this work was to investigate the effect of covalent labelling on the physico-chemical properties of β-casein (β-CN) in solution and in emulsions stabilized by β-CN and whey protein isolate (WPI). β-CN was covalently labelled by 5-(and 6)-carboxytetramethylrhodamine, succinimidyl ester (NHS-Rhodamine). The effect of conjugating β-CN with NHS-Rhodamine on the spectroscopic properties of labelled β-CN (β-CNlabelled) was examined. No significant difference in interfacial tension (p > 0.05) was found between mixture of WPI and β-CNlabelled (0.5% w/w WPI/β-CNlabelled) and of WPI and β-CN (0.5% w/w WPI/β-CN) in 10 mM phosphate buffer (pH 7.0) at 20 °C. Oil-in-water emulsions stabilized with either WPI/β-CN or WPI/β-CNlabelled (0.5% w/w) were also investigated using laser-light scattering, analytical centrifugation, rheometry and CLSM. It was shown that labelling had no significant effect on the physico-chemical properties of emulsions (p > 0.05) in terms of droplet size, creaming stability, viscosity or zeta-potential. Confocal micrographs of emulsions made with WPI/β-CNlabelled showed that both β-CN and whey proteins could be observed simultaneously, and were co-localized at the surface of fat globules. Furthermore, it was found through image analysis that β-CN produced a thicker interfacial layer than WPI

Influence of emulsifier type on the spray-drying properties of model infant formula emulsions

The objective of this study was to compare the drying performance and physicochemical properties of model infant formula (IF) emulsions containing 43, 96 and 192 g L−1 protein, oil and maltodextrin (MD), respectively, prepared using different emulsifier systems. Emulsions were stabilised using either whey protein isolate (WPI), whey protein hydrolysate (WPH; DH 8%), WPH + CITREM (9 g L−1), WPH + lecithin (5 g L−1) or WPH conjugated with maltodextrin (DE 12) (WPH-MD). Homogenised emulsions had 32% solids content and oil globules with mean volume diameter WPH + LEC > WPH > WPH- MD > WPI, WPI > WPH > WPH- MD > WPH + LEC > WPH + CIT and WPH- MD > WPI > WPH > WPH + LEC > WPH + CIT, respectively. Additionally, differences in wettability, surface topography and oil globule distribution within the powder matrix and in reconstituted powders were linked to the emulsifier system used. Inclusion of the WPH-MD conjugate in the formulation of IF powder significantly improved drying behaviour and physicochemical properties of the resultant powder, as evidenced by lowest powder build-up during drying and greatest emulsion quality on reconstitution, compared to the other model formula systems

In vitro digestion of protein-enriched restructured beef steaks with pea protein isolate, rice protein and lentil flour following sous vide processing

peer-reviewedThe effect of plant protein inclusion in cooked meat upon in vitro gastro-intestinal (GI) digestion was investigated. Pea protein isolate, rice protein and lentil flour were used to increase the protein content in a meat model system restructured using two transglutaminase enzymes [Activa®EB (TG) and Transgluseen™-M (TS)]. Restructured beef steaks were subjected to simulated GI digestion using the static INFOGEST method. Samples taken at different digestion times were analysed using SDS-PAGE, size exclusion-HPLC, free amino acid analysis and microscopy. SDS-PAGE analysis revealed significant protein hydrolysis during GI digestion. Most soluble peptides had a molecular weight smaller than 500 Da, corresponding to peptides of <5 amino acids, regardless of food treatment. The amounts of released, free amino acids isoleucine, lysine, phenylalanine and valine were higher (P < 0.05) in lentil-enriched restructured beef steaks following GI digestion. Confocal laser scanning microscopy (CSLM) revealed pronounced aggregation in digested samples. In vitro digestates of protein-enriched restructured beef steaks showed lower production of small molecular weight peptides. This study demonstrated how the bioaccessibility of protein-enriched restructured beef steaks are influenced by formulation and processing

Dairy Ingredients for the Baking Industry.

End of Project ReportShortenings (baking fats), microencapsulated using dairy ingredients and milk protein hydrolysates, were produced for testing in a variety of baked products. The powders were evaluated for their functionality as powdered baking fats, as potential replacers of synthetic emulsifiers, as ingredients capable of improving baking performance or as potential health-enhancing ingredients. These studies provide the technology for the dairy industry to enter the specialised food ingredients sector with a siftable, non-greasy, free-flowing powdered fat for the baking industry.Department of Agriculture, Food and the Marin

Optimisation of plant protein and transglutaminase content in novel beef restructured steaks for older adults by central composite design

peer-reviewedWith the goal of optimising a protein-enriched restructured beef steak targeted at the nutritional and chemosensory requirements of older adults, technological performance of thirty formulations, containing plant-based ingredients, pea protein isolate (PPI), rice protein (RP) and lentil flour (LF) with transglutaminase (TG) to enhance binding of meat pieces, were analysed. Maximal protein content of 28% in cooked product was achieved with PPI, RP and LF. Binding strength was primarily affected by TG, while textural parameters were improved with LF inclusion. Optimal formulation (F) to obtain a protein-enriched steak with lowest hardness values was achieved with TG (2%), PPI (8%), RP (9.35%) and LF (4%). F, F1S (optimal formulation 1 with added seasoning) and control restructured products (not containing plant proteins or seasonings) were scored by 120 consumers' aged over-65 years. Controls were most preferred (P < .05), while F1S were least liked by the older consumers. Consumer testing suggests further refinement and optimisation of restructured products with plant proteins should be undertaken.This research was funded by the FIRM programme administered under the Irish Department of Agriculture, Food and the Marine, Meat4Vitality: Enhancement of texture, flavour and nutritional value of meat products for older adults and the Walsh Fellowship Scheme (11/F/045)