Functionality of glycomacropeptide glycated with lactose and maltodextrin.

The Maillard reaction (MR), under proper environmental conditions, has been used to improve protein functionality. In the present work, 2 high temperatures (50-80°C) and water activity (Aw; 0.45-0.67) were used to promote exogenous glycosylation of glycomacropeptide (GMP) while minimizing processing times (0, 8, 24, 48, and 96 h at 50°C; 0, 2, 4, 8, and 24 h at 80°C). Maltodextrin, a polysaccharide commonly used in the food industry as a functional ingredient, was used as a reducing sugar, and compared with lactose, a native milk sugar. The progression of MR was evaluated by tracking changes in molecular weight using SDS-PAGE, the formation of Amadori compounds, and browning. Aqueous glycosylated GMP solutions (5 to 20% wt/vol) were tested for solubility, rheological properties, and foam formation. As expected, MR progression was faster with Aw = 0.67 and 80°C. Glycosylated GMP powders showed no change in their solubility after MR. However, the apparent viscosity (γ˙=30s-1) of the 20% wt/vol suspensions exhibited a slight increase when GMP was glycosylated with maltodextrin for 24 h at 80°C, and a 2-log increase when GMP was glycosylated with lactose, with a high browning development in both cases. The foam expansion index of the resuspended glycosylated powders was increased by between 25 and 66% compared with the nonglycosylated powders. Better foam stability (approximately 2 h) and no browning development were observed for GMP glycosylated with maltodextrin for 2 h at Aw = 0.67 and 80°C. The results show that GMP has undergone further glycosylation by means of controlled MR, which improves viscosity and foaming index without negatively affecting solubility. These preliminary studies provide a basis for the future creation of a new ingredient with GMP and reducing sugars.

Effect of high-pressure-jet processing on the physiochemical properties of low-fat ice cream mix.

The objective of this study was to use high-pressure-jet (HPJ) processing to produce functional properties in a low-fat (4.5% fat) ice cream mix similar to those seen when emulsifiers are used. Ice cream mix or serum (nonfat portion of the ice cream mix) were subjected to 200 or 400 MPa HPJ processing and compared with a non-HPJ-treated control. A similar non-HPJ-treated formulation but containing polysorbate 80 (0.075% wt/wt) was also used as a control. The mix samples were characterized in terms of their particle size, density, flow properties, stability, crystallization kinetics, and fat-protein interactions. The sample from the mix subjected to 400 MPa HPJ processing (HPJ-M-400) had increased consistency coefficient (5°C; 228 ± 102.7 mPa·s) and particle size (D[4,3]; 16.0 ± 2.5 µm) compared with the non-HPJ-treated control sample, with viscosity and particle size (volume-moment mean diameter, D[4,3]) values of 7.5 ± 0.4 mPa·s and 0.50 ± 0.1 µm, respectively. These differences were attributed to an increase in casein-fat interactions and casein-casein interactions caused by the 400 MPa HPJ treatment, which were observed using confocal scanning laser microscopy and inferred from an increase in protein and fat concentrations in the sediment after ultracentrifugation. Interestingly, the density of HPJ-M-400 was also lower (0.79 ± 0.17 g/mL) than that of the control (1.04 ± 0.00 g/mL) because bubbles were trapped within these complexes. The large casein-fat complexes formed in the HPJ-M-400 sample also appeared to act as steric barriers that slowed ice crystal growth during quiescent freezing. The alterations in physiochemical properties and apparent ice crystal growth induced by the 400 MPa treatment of low-fat ice cream mix have many potential applications, including clean-label confections.

Assessing constituent volumes and morphology of bovine casein micelles using cryo-electron tomography.

We investigated the applicability of cryo-electron tomography as a method to quantify changes in the major constituents of casein micelles (i.e., casein proteins, putative colloidal calcium phosphate nanoclusters, and serum-filled voids and channels) in response to their environment. Skim milk diluted 20-fold in milk serum was used for this study. Tomograms were generated for multiple casein micelles at 2 different pH values (6.7 and 6.0) and pixel intensity thresholds were identified for each constituent. The volume of each constituent was determined using these thresholds and expressed as a fraction of micelle volume. At the given dilution, a significant decrease in the volume fractions of casein proteins (∼37%) and putative colloidal calcium phosphate nanoclusters (∼67%) was observed with the reduction of pH from 6.7 to 6.0. Assessment of casein micelle fraction obtained by ultracentrifugation of corresponding skim milk samples produced comparable results. When using such an approach, the imaging conditions, denoising methods, and thresholding approaches used can all affect the precision of the measurements, but the overall trends in constituent volumes are able to be tracked. The primary advantage of using cryo-electron tomography is that analysis can be done at the level of individual micelles, within a 3-dimensional morphological context. This workflow paves the way for high-throughput exploration of milk micelles and how their environment shapes their composition and structure.

Chromatographic fractionation and molecular mass characterization of Cercidium praecox (Brea) gum.

BACKGROUND: Brea gum (BG) is an exudate from the Cercidium praecox tree that grows in semi-arid regions of Argentina. Some previous studies on BG have shown physicochemical characteristics and functional features similar to those of gum arabic. However, there is a need to elucidate the molecular structure of BG to understand the functionality. In this sense, BG was fractionated using hydrophobic interaction chromatography and the obtained fractions were analyzed by size exclusion chromatography. RESULTS: Analysis of the fractions showed that the bulk of the gum (approx. 84% of the polysaccharides) was a polysaccharide of 2.79 × 10(3) kDa. The second major fraction (approx. 16% of the polysaccharides) was a polysaccharide-protein complex with a molecular mass of 1.92 × 10(5) kDa. A third fraction consisted of protein species with a wide range of molecular weights. The molecular weight distribution of the protein fraction was analyzed by size exclusion chromatography. Comparison of the elution profiles of the exudates in native and reducing conditions revealed that some of the proteins were forming aggregates through disulfide bridges in native conditions. Further analysis of the protein fraction by SDS-PAGE showed proteins with molecular weight ranging from 6.5 to 66 kDa. CONCLUSIONS: The findings showed that BG consists of several fractions with heterogeneous chemical composition and polydisperse molecular weight distributions. © 2016 Society of Chemical Industry.

Choline and choline metabolite patterns and associations in blood and milk during lactation in dairy cows.

Milk and dairy products are an important source of choline, a nutrient essential for human health. Infant formula derived from bovine milk contains a number of metabolic forms of choline, all contribute to the growth and development of the newborn. At present, little is known about the factors that influence the concentrations of choline metabolites in milk. The objectives of this study were to characterize and then evaluate associations for choline and its metabolites in blood and milk through the first 37 weeks of lactation in the dairy cow. Milk and blood samples from twelve Holstein cows were collected in early, mid and late lactation and analyzed for acetylcholine, free choline, betaine, glycerophosphocholine, lysophosphatidylcholine, phosphatidylcholine, phosphocholine and sphingomyelin using hydrophilic interaction liquid chromatography-tandem mass spectrometry, and quantified using stable isotope-labeled internal standards. Total choline concentration in plasma, which was almost entirely phosphatidylcholine, increased 10-times from early to late lactation (1305 to 13,535 µmol/L). In milk, phosphocholine was the main metabolite in early lactation (492 µmol/L), which is a similar concentration to that found in human milk, however, phosphocholine concentration decreased exponentially through lactation to 43 µmol/L in late lactation. In contrast, phosphatidylcholine was the main metabolite in mid and late lactation (188 µmol/L and 659 µmol/L, respectively), with the increase through lactation positively correlated with phosphatidylcholine in plasma (R2 = 0.78). Unlike previously reported with human milk we found no correlation between plasma free choline concentration and milk choline metabolites. The changes in pattern of phosphocholine and phosphatidylcholine in milk through lactation observed in the bovine suggests that it is possible to manufacture infant formula that more closely matches these metabolites profile in human milk.

Inactivation of Alicyclobacillus acidoterrestris using high pressure homogenization and dimethyl dicarbonate.

Vegetative cells and spores of five strains of Alicyclobacillus acidoterrestris (N-1100, N-1108, N-1096, SAC, and OS-CAJ) were screened for their sensitivity to high pressure homogenization (HPH, 0 to 300 MPa) in Bacillus acidoterrestris thermophilic broth. The most and least resistant strains, SAC and OS-CAJ, respectively, were further tested for their sensitivity to inactivation or growth inhibition by dimethyl dicarbonate (DMDC, 250 ppm). The combined effects of HPH and DMDC were then evaluated against SAC spores over a 24-h period after treatment. HPH alone significantly inactivated (P < 0.05) vegetative cells of all five strains. SAC vegetative cells were least affected by HPH, with only about a 0.5-log reduction after the 300-MPa treatment. Spores were not significantly reduced by HPH for any of the five strains. DMDC reduced the initial vegetative cell population by 2 log CFU/ml and significantly increased the time to reach stationary phase. For spores, a 0.5-log decrease from the initial spore population was achieved and growth was not significantly delayed. No significant difference was found between the two strains. Treatment with DMDC plus HPH slightly enhanced the inactivation effect over a 24-h period compared with treatment with HPH alone, but these differences were statistically inconsistent. Although HPH and DMDC treatments may help control vegetative cells of A. acidoterrestris, these treatments may not provide adequate overall control. Neither treatment, alone or in combination, is very effective against spores.

Survival and inactivation of human norovirus surrogates in blueberry juice by high-pressure homogenization.

Human noroviruses (HNoV) have been implicated in gastrointestinal outbreaks associated with fresh produce, juices, and ready-to-eat foods. In order to determine the risk of HNoV transmission by contaminated blueberry juice, survival characteristics of cultivable HNoV surrogates (murine norovirus, MNV-1; feline calicivirus, FCV-F9; and bacteriophage MS2) in blueberry juice (pH = 2.77) after 0, 1, 2, 7, 14, and 21 days at refrigeration temperatures (4°C) were studied. High-pressure homogenization (HPH) was studied as a novel processing method for noroviral surrogate inactivation in blueberry juice. Blueberry juice or phosphate-buffered saline (PBS; pH 7.2 as control) was inoculated with each virus, stored over 21 days at 4°C or subjected to HPH, and plaque assayed. FCV-F9 (∼5 log(10) PFU/mL) was undetectable after 1 day in blueberry juice at 4°C. MNV-1 (∼4 log(10) PFU/ml) showed minimal reduction (1 log(10) PFU/mL) after 14 days, with greater reduction (1.95 log(10) PFU/mL; p < 0.05) after 21 days in blueberry juice at 4°C. Bacteriophage MS2 (∼6 log(10) PFU/mL) showed significant reduction (1.93 log(10) PFU/mL; p < 0.05) after 2 days and was undetectable after 7 days in blueberry juice at 4°C. FCV-F9 remained viable in PBS for up to 21 days (2.28 log(10) PFU/mL reduction), while MNV-1 and MS2 survived after 21 days (1.08 and 0.56 log(10) PFU/mL reduction, respectively). Intriguingly, FCV-F9 and bacteriophage MS2 showed reduction after minimal homogenization pressures in blueberry juice (pH = 2.77), possibly due to the combination of juice pH, juice components, and mechanical effects. MNV-1 in blueberry juice was only slightly reduced at 250 (0.33 log(10) PFU/mL) and 300 MPa (0.71 log(10) PFU/mL). Virus surrogate survival in blueberry juice at 4°C correlates well with the ease of HNoV transmission via juices. HPH for viral inactivation in juices is dependent on virus type, and higher homogenization pressures may be needed for MNV-1 inactivation.

Antimicrobial activity of Yerba Mate (Ilex paraguariensis) aqueous extracts against Escherichia coli O157:H7 and Staphylococcus aureus.

UNLABELLED: Bioactive compounds from natural plant sources are becoming increasingly important to the food industry. Ilex paraguariensis is used in the preparation of a widely popular tea beverage (Yerba Mate) in the countries of Uruguay, Paraguay, Argentina, and Brazil. In this study, extracts of 4 brands of commercial tea, derived from the holly plant species, Ilex paraguariensis, were evaluated for their ability to inhibit or inactivate bacterial foodborne pathogens. The ultimate goal was to evaluate potential use of the extracts in commercial applications. Dialyzed aqueous extracts were screened for antimicrobial activity against Escherichia coli O157:H7 and Staphylococcus aureus. S. aureus was found to be the more sensitive to extracts than E. coli O157:H7. Minimum bactericidal concentrations (MBCs) were determined to be approximately 150 to 800 µg/mL and 25 to 50 µg/mL against E. coli O157:H7 and S. aureus, respectively. A Uruguayan brand had reduced activity against E. coli O157:H7 compared to the Argentinean brands tested. It was concluded that Yerba Mate could be used as a potential antimicrobial in foods and beverages against these pathogenic bacteria. PRACTICAL APPLICATION: Soluble extracts from Yerba Mate are natural antimicrobials that can be incorporated into food products to achieve longer shelf life.

Effect of high hydrostatic pressure and whey proteins on the disruption of casein micelle isolates.

High hydrostatic pressure disruption of casein micelle isolates was studied by analytical ultracentrifugation and transmission electron microscopy. Casein micelles were isolated from skim milk and subjected to combinations of thermal treatment (85 degrees C, 20 min) and high hydrostatic pressure (up to 676 MPa) with and without whey protein added. High hydrostatic pressure promoted extensive disruption of the casein micelles in the 250 to 310 MPa pressure range. At pressures greater than 310 MPa no further disruption was observed. The addition of whey protein to casein micelle isolates protected the micelles from high hydrostatic pressure induced disruption only when the mix was thermally processed before pressure treatment. The more whey protein was added (up to 5 g/l) the more the protection against high hydrostatic pressure induced micelle disruption was observed in thermally treated samples subjected to 310 MPa.