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

Encapsulant 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.

Reduced-fat Cheddar and Swiss-type cheeses harboring exopolysaccharide-producing probiotic Lactobacillus mucosae DPC 6426.

Exopolysaccharide-producing Lactobacillus mucosae DPC 6426 was previously shown to have promising hypocholesterolemic activity in the atherosclerosis-prone apolipoprotein-E-deficient (apoE(-/-)) murine model. The aim of this study was to investigate the suitability of reduced-fat Cheddar and Swiss-type cheeses as functional (carrier) foods for delivery of this probiotic strain. All cheeses were manufactured at pilot-scale (500-L vats) in triplicate, with standard commercially available starters: for Cheddar, Lactococcus lactis; and for Swiss-type cheese, Streptococcus thermophilus, Lactobacillus helveticus, and Propionibacterium freudenreichii. Lactobacillus mucosae DPC 6426 was used as an adjunct culture during cheese manufacture, at a level of ~10(6) cfu·mL(-1) cheese milk (subsequently present in the cheese curd at>10(7) cfu·g(-1)). The adjunct strain remained viable at >5×10(7) cfu·g(-1) in both Swiss-type and Cheddar cheeses following ripening for 6 mo. Sensory analysis revealed that the presence of the adjunct culture imparted a more appealing appearance in Swiss-type cheese, but had no significant effect on the sensory characteristics of Cheddar cheeses. Moreover, the adjunct culture had no significant effect on cheese composition, proteolysis, pH, or instrumentally quantified textural characteristics of Cheddar cheeses. These data indicate that low-fat Swiss-type and Cheddar cheeses represent suitable food matrices for the delivery of the hypocholesterolemic Lactobacillus mucosae DPC 6426 in an industrial setting.

Characterization of a bovine isolate Lactobacillus mucosae DPC 6426 which produces an exopolysaccharide composed predominantly of mannose residues.

AIMS: To characterize Lactobacillus strains with EPS-producing ability compared with non-EPS-producing lactobacilli of the same species for technological performance including simulated gastrointestinal tract (GIT) conditions. METHODS AND RESULTS: Characterization of EPS-producing Lactobacillus mucosae DPC 6426 in detail based on 16S rRNA sequencing, and EPS production using scanning electron and atomic force microscopy. The EPS was found to consist of mannosyl residues, with mannose, glucose and galactose found to be the major sugar residues present in an approximate ratio of 3: 2: 2. The strain was compared to non-EPS-producing Lact. mucosae DPC 6420 following exposure to salt, bile, acid and heat stresses. Lact. mucosae DPC 6426 exhibited twofold increased (P < 0·05) survival during 120-min exposure to 5 mol NaCl, threefold increased survival during 90-min exposure to 0·7% (w/v) bile (P < 0·05), threefold increased survival when exposed to simulated gastric juice (P < 0·001) for 10 min and fivefold increased survival during 60-min exposure to HCl (P < 0·01) compared with Lact. mucosae DPC 6420. Furthermore, Lact. mucosae DPC 6426 was found to be more heat tolerant (P < 0·001) compared with Lact. mucosae DPC 6420 during 30-min exposure to 55°C. CONCLUSIONS: These data indicate that the EPS-producing Lact. mucosae DPC 6426 exhibits technological and biological robustness compared with a non-EPS-producing Lact. mucosae strain. SIGNIFICANCE AND IMPACT OF THE STUDY: The data implicate the potential suitability of EPS-producing Lact. mucosae DPC 6426 in food applications and/or as a probiotic culture.

Application of whey protein micro-bead coatings for enhanced strength and probiotic protection during fruit juice storage and gastric incubation.

CONTEXT: Coated whey protein micro-beads may improve probiotic protection and provide delayed cell-release mechanisms. OBJECTIVE: Lactobacillus rhamnosus GG was encapsulated in whey protein micro-beads by droplet extrusion with coating via electrostatic deposition: primary-polysaccharide and secondary-whey protein. MATERIALS AND METHODS: Storage studies were performed in cranberry and pomegranate juice (pH 2.4; 28 days; 4 and 25°C) followed by simulated ex vivo porcine gastric (pH 1.6) and intestinal (pH 6.6) digestion. RESULTS AND DISCUSSION: After storage and simulated gastro-intestinal digestion, free cells, cells suspended in protein and cells encapsulated in alginate micro-beads, illustrated complete probiotic mortality, while coated micro-beads enhanced probiotic viability after juice storage (8.6 ± 0.1 log(10)CFUmL(-1)). Beads also showed significant binding of hydrophobic molecules. Coated micro-beads illustrated high gastric survival (9.5 ± 0.1 log(10)CFUmL(-1)) with 30 min delayed intestinal release relative to non-coated micro-beads. CONCLUSIONS: Micro-bead coatings could be applied in delayed cell-release for targeted intestinal probiotic delivery.

Effect of exopolysaccharide produced by isogenic strains of Lactococcus lactis on half-fat Cheddar cheese.

Fat-reduced cheeses often suffer from undesirable texture, flavor, and cooking properties. Exopolysaccharides (EPS) produced by starter strains have been proposed as a mechanism to increase yield and to improve the texture and cooking properties of reduced-fat cheeses. The objective of this work was to assess the influence of an exopolysaccharide on the yield, texture, cooking properties, and quality of half-fat Cheddar cheese. Two pilot-scale half-fat Cheddar cheeses were manufactured using single starters of an isogenic strain of Lactococcus lactis ssp. cremoris (DPC6532 and DPC6533) that differed in their ability to produce exopolysaccharide. Consequently, any differences detected between the cheeses were attributed to the presence of the exopolysaccharide. The results indicated that cheeses made with the exopolysaccharide-producing starter had an 8.17% increase in actual cheese yield (per 100 kg of milk), a 9.49% increase in moisture content, increase in water activity and water desorption rate at relative humidities

A new cryo-STEM method for imaging food colloids in the scanning electron microscope.

A new cryo-scanning transmission electron microscopy (cryo-STEM) technique for imaging casein micelles in a field emission scanning electron microscope is presented. Thin films of micellar casein suspensions on lacey carbon grids were prepared using a modified sample holder developed by Gatan UK. Bright and dark field images were obtained at -135°C showing casein micelles in their frozen hydrated state and in the size range 30-500 nm. Results were compared favorably with published images of casein micelles obtained with conventional cryo-transmission electron microscopy, suggesting that cryo-STEM is a useful alternative technique for visualizing food colloids close to their native state.

The effect of natural cheddar cheese ripening on the functional and textural properties of the processed cheese manufactured therefrom.

Cheddar cheese ripened at 8 degrees C was sampled at 7, 14, 28, 56, 112, and 168 d and subsequently used for the manufacture of processed cheese. The cheddar cheese samples were analyzed throughout ripening for proteolysis while the textural and rheological properties of the processed cheeses (PCs) were studied. The rate of proteolysis was the greatest in the first 28 d of cheddar cheese ripening but began to slow down as ripening progressed from 28 to 168 d. A similar trend was observed in changes to the texture of the PC samples, with the greatest decrease in hardness and increase in flowability being in the first 28 d of ripening. Confocal scanning laser microscopy showed that the degree of emulsification in the PC samples increased as the maturity of the cheddar cheese ingredient increased from 7 to 168 d. This increased emulsification resulted in a reduction in the rate of softening in the PC in samples manufactured from cheddar cheese bases at later ripening times. Multivariate data analysis was performed to summarize the relationships between proteolysis in the cheddar cheese bases and textural properties of the PC made therefrom. The proportion of alpha(s)(1)-casein (CN) in the cheddar cheese base was strongly correlated with hardness, adhesiveness, fracturability, springiness, and storage modulus values for the corresponding PC. Degradation of alpha(s) (1)-CN was the proteolytic event with the strongest correlation to the softening of PC samples, particularly those manufactured from cheddar cheese in the first 28 d of ripening.

Effect of pH and calcium concentration on some textural and functional properties of mozzarella cheese.

The effects of Ca concentration and pH on the composition, microstructural, and functional properties of Mozzarella cheese were studied. Cheeses were made using a starter culture (control) or by direct acidification of the milk with lactic acid or lactic acid and glucono-delta-lactone. In each of three trials, four cheeses were produced: a control, CL, and three directly-acidified cheeses, DA1, DA2, and DA3. The cheeses were stored at 4 degrees C for 70 d. The Ca content and pH were varied by altering the pH at setting, pitching, and plasticization. The mean pH at 1 d and the Ca content (mg/g of protein) of the various cheeses were: CL, 5.42 and 27.7; DA1, 5.96 and 21.8; DA2, 5.93 and 29.6; DA3, 5.58 and 28.7. For cheeses with a high pH (i.e., approximately 5.9), reducing the Ca content from 29.6 to 21.8 mg/g of protein resulted in a significant decrease in the protein level and increases in the moisture content and mean level of nonexpressible serum (g/g of protein). Reducing the Ca concentration also resulted in a more swollen, hydrated para-casein matrix at 1 d. The decrease in Ca content in the high-pH cheeses coincided with increases in the mean stretchability and flowability of the melted cheese over the 70-d storage period. The fluidity of the melted cheese also increased when the Ca content was reduced, as reflected by a lower elastic shear modulus and a higher value for the phase angle, delta, of the melted cheese, especially after storage for <12 d. The melt time, flowability, and stretchability of the low-Ca, high-pH DA1 cheese at 1 d were similar to those for the CL cheese after storage for > or = 12 d. In contrast, the mean values for flowability and stretchability of the high-pH, high-Ca DA2 cheese over the 70-d period were significantly lower than those of the CL cheese. Reducing the pH of high-Ca cheese (27.7 to 29.6 mg/g of protein) from -5.95 to 5.58 resulted in higher flowability, stretchability, and fluidity of the melted cheese. For cheeses with similar pH and Ca concentration, the method of acidification had little effect on composition, microstructure, flowability, stretchability, and fluidity of the melted cheese.

Development and application of confocal scanning laser microscopy methods for studying the distribution of fat and protein in selected dairy products.

Confocal scanning laser microscopy (CSLM) methods were developed to identify fat and protein in cheeses milk chocolate and milk powders. Various fluorescent probes were assessed for their ability to label fat or protein in selected food products in situ. Dual labelling of fat and protein was made possible by using mixtures of probes. Selected probes and probe mixtures were then used to study (a) structure development of Mozzarella cheese during manufacture and ripening, and (b)) the distribution of fat and protein in milk chocolate made with milk powders containing varying levels of free fat. Microstructural changes in the protein and fat phases of Mozzarella cheese were observed at each major step in processing. Aggregation of renneted micelles occurred during curd formation; this was followed by amalgamation of the para-casein into linear fibres during plasticization. Following storage, the protein phase of the Mozzarella became more continuous; entrapping and isolating fat globules. Chocolate made with a high free-fat spray-dried powder blend showed a homogeneous fat distribution, similar to that of chocolate made with roller-dried milk. Chocolate made with whole milk powder containing 10 g free fat/100 fat showed a non-homogeneous fat distribution with some fat occluded within milk protein particles. These differences in fat distribution were related to Casson yield value and Casson viscosity of the chocolates.

Ethanol-dependent heat-induced dissociation of casein micelles.

The dissociation of casein micelles when heated to approximately 65 degrees C in the presence of ethanol [1:1 mixture (v/v) of milk and 65% (w/w) aqueous ethanol] was investigated using L* values and transmission measurements. Mixtures of milk and ethanol became transparent on heating, which suggests dissociation of casein micelles. Results of experiments using confocal laser scanning microscopy, light scattering (static and dynamic), and dialysis to examine the changes of milk during heating in the presence of ethanol supported the assertion that such treatments result in dissociation of casein micelles, as did studies of model beta-casein micellar systems.

Direct in situ viability assessment of bacteria in probiotic dairy products using viability staining in conjunction with confocal scanning laser microscopy.

The viability of the human probiotic strains Lactobacillus paracasei NFBC 338 and Bifidobacterium sp. strain UCC 35612 in reconstituted skim milk was assessed by confocal scanning laser microscopy using the LIVE/DEAD BacLight viability stain. The technique was rapid (<30 min) and clearly differentiated live from heat-killed bacteria. The microscopic enumeration of various proportions of viable to heat-killed bacteria was then compared with conventional plating on nutrient agar. Direct microscopic enumeration of bacteria indicated that plate counting led to an underestimation of bacterial numbers, which was most likely related to clumping. Similarly, LIVE/DEAD BacLight staining yielded bacterial counts that were higher than cell numbers obtained by plate counting (CFU) in milk and fermented milk. These results indicate the value of the microscopic approach for rapid viability testing of such probiotic products. In contrast, the numbers obtained by direct microscopic counting for Cheddar cheese and spray-dried probiotic milk powder were lower than those obtained by plate counting. These results highlight the limitations of LIVE/DEAD BacLight staining and the need to optimize the technique for different strain-product combinations. The minimum detection limit for in situ viability staining in conjunction with confocal scanning laser microscopy enumeration was approximately 10(8) bacteria/ml (equivalent to approximately 10(7) CFU/ml), based on Bifidobacterium sp. strain UCC 35612 counts in maximum-recovery diluent.

Comparative survival rates of human-derived probiotic Lactobacillus paracasei and L. salivarius strains during heat treatment and spray drying.

Spray drying of skim milk was evaluated as a means of preserving Lactobacillus paracasei NFBC 338 and Lactobacillus salivarius UCC 118, which are human-derived strains with probiotic potential. Our initial experiments revealed that NFBC 338 is considerably more heat resistant in 20% (wt/vol) skim milk than UCC 118 is; the comparable decimal reduction times were 11.1 and 1.1 min, respectively, at 59 degrees C. An air outlet temperature of 80 to 85 degrees C was optimal for spray drying; these conditions resulted in powders with moisture contents of 4.1 to 4.2% and viable counts of 3.2 x 10(9) CFU/g for NFBC 338 and 5.2 x 10(7) CFU/g for UCC 118. Thus, L. paracasei NFBC 338 survived better than L. salivarius UCC 118 during spray drying; similar results were obtained when we used confocal scanning laser microscopy and LIVE/DEAD BacLight viability staining. In addition, confocal scanning laser microscopy revealed that the probiotic lactobacilli were located primarily in the powder particles. Although both spray-dried cultures appeared to be stressed, as shown by increased sensitivity to NaCl, bacteriocin production by UCC 118 was not affected by the process, nor was the activity of the bacteriocin peptide. The level of survival of NFBC 338 remained constant at approximately 1 x 10(9) CFU/g during 2 months of powder storage at 4 degrees C, while a decline in the level of survival of approximately 1 log (from 7.2 x 10(7) to 9.5 x 10(6) CFU/g) was observed for UCC 118 stored under the same conditions. However, survival of both Lactobacillus strains during powder storage was inversely related to the storage temperature. Our data demonstrate that spray drying may be a cost-effective way to produce large quantities of some probiotic cultures.