Size, shape, and identity of surface crystals and their relationship to sensory perception of grittiness in soft smear-ripened cheeses.

Soft smear-ripened cheeses undergo extensive surface crystallization and radial demineralization of calcium, magnesium, and phosphorus, which likely contributes to radial softening during ripening. Furthermore, anecdotal evidence suggests that grittiness is a common characteristic of smear-ripened cheeses. The primary aims of the present study were to evaluate the intensity of perceived grittiness while assessing other key sensory attributes in US artisanal and European protected designation of origin smear-ripened cheeses, and to relate perceived grittiness to the size, shape, and identity of crystals present in the cheese surface smears. Fully ripened wheels of 24 different varieties of smear-ripened cheeses, 16 produced in the United States and 8 in the European Union, were obtained from retail sources. A trained sensory panel (n = 12) was employed to evaluate intensity of grittiness. Crystals present in the cheese smears were identified by powder X-ray diffractometry and polarized light microscopy, and further evaluated in polarized light microscopy micrographs by image analysis for size and shape characteristics. Mean sensory scores for the 24 cheeses ranged from no perceived grittiness to easily identifiable grittiness. Surface crystals included ikaite, struvite, calcite, and brushite, and mean crystal length and area ranged among cheeses from 27 to 1,096 µm, and 533 to 213,969 µm2, respectively. Panel threshold for grittiness occurred at a mean crystal length of about 66 µm and mean crystal area of about 2,913 µm2. Cheeses with mean values at or below these thresholds displayed negligible perceived grittiness. In contrast, for cheeses with mean values above these thresholds, the mean sensory scores for grittiness were highly correlated with mean crystal length and crystal area (r = 0.93 and 0.96, respectively). Results suggest that surface crystals in soft smear-ripened cheeses influence sensory perception of texture in complex ways that likely include radial softening and grittiness development. A better understanding of factors that govern surface crystal formation may lead to improved control over crystallization and more consistent cheese texture.

Characterization and identification of surface crystals on smear-ripened cheese by polarized light microscopy.

Surface crystallization and radial demineralization of Ca, P, and Mg occur in smear-ripened cheese. Furthermore, crystals of ikaite, struvite, calcite, and brushite have been identified in cheese smears by powder X-ray diffractometry (PXRD), and ikaite and struvite exist in smears as single crystals. Polarized light microscopy (PLM) is a simple, inexpensive, and well-established method in geology to detect and identify single crystals. However, use of PLM to identify cheese crystals has not been reported previously. The specific objectives of this research were (1) to identify crystals in cheese smears using selected PLM criteria; (2) to compare identification by PLM against PXRD; and (3) to develop and evaluate a novel treatment for smear material to improve crystal analyses by both PLM and PXRD. Duplicate wheels of 4 cheeses produced by different manufacturers were obtained from retail sources. Scrapings of surface smears were prepared and analyzed by PLM and PXRD by previously described methods. Crystals were categorized by PLM based on angle of extinction (AE), birefringence behavior under crossed polarizers and quartz filters, and size and shape (circularity) by image analysis. Crystals observed by PLM fell almost exclusively into 2 readily differentiated groups based on birefringence behavior and estimated angle of extinction. Group 1 (n = 18) were highly birefringent with AE = 88-92°, whereas group 2 (n = 28) had no birefringence with AE = 13-26°. Group 2 crystals were significantly larger and more circular than group 1 crystals. Group 1 and 2 were identified as struvite and ikaite, respectively, based on known birefringence and AE characteristics. Struvite was identified in all 4 cheeses by PLM but in only 3 cheeses by PXRD. Ikaite was identified in 3 cheeses by PLM but in only 2 cheeses by PXRD. These discrepancies occurred because the smear scrapings from 1 cheese contained excessive amorphous matter that caused extreme background noise, potentially obscuring diffractogram peaks that may have been present. To minimize noise, smear scrapings were dispersed in aqueous NaOH (pH 10) before analyses, which resulted in consistent results by PXRD and PLM. The method also rendered high-quality images by PLM. Data suggest that PLM may offer a simple and inexpensive means to identify struvite, ikaite, and possibly other single crystals in cheese smears.

Crystal fingerprinting: elucidating the crystals of Cheddar, Parmigiano-Reggiano, Gouda, and soft washed-rind cheeses using powder x-ray diffractometry.

Crystals in cheese may be considered defects or positive features, depending on the variety and mode of production (industrial, artisanal). Powder x-ray diffractometry (PXRD) offers a simple means to identify and resolve complex combinations of crystals that contribute to cheese characteristics. The objective of the present research was to demonstrate the application of PXRD to study crystals from a range of different cheese types, specifically Cheddar, Parmigiano-Reggiano, Gouda, and soft washed-rind (smear ripened) cheeses. In studies of Parmigiano-Reggiano and long-aged Gouda, PXRD has confirmed that hard (crunchy) crystals that form abundantly within these cheeses consist of tyrosine. Furthermore, PXRD has tentatively identified the presence of an unusual form of crystalline leucine in large (up to 6 mm in diameter) spherical entities, or "pearls", that occur abundantly in 2-year-old Parmigiano Reggiano and long-aged Gouda cheeses, and on the surface of rindless hard Italian-type cheese. Ongoing investigations into the nature of these "pearls" are providing new insight into the roles that crystals play in the visual appearance and texture of long-aged cheeses. Crystals also sometimes develop profusely in the eyes of long-aged Gouda, which have been shown by PXRD to consist of tyrosine and the aforementioned presumptive form of crystalline leucine. Finally, crystals have been shown by PXRD to form in the smears of soft washed-rind cheeses. These crystals may be associated in some cheeses with gritty mouth feel and with zonal body softening that occurs during ripening. Heightened interest in artisanal cheeses highlights the need to better understand crystals and their contributions to cheese characteristics.

Powder X-ray diffraction can differentiate between enantiomeric variants of calcium lactate pentahydrate crystal in cheese.

Powder X-ray diffraction has been used for decades to identify crystals of calcium lactate pentahydrate in Cheddar cheese. According to this method, diffraction patterns are generated from a powdered sample of the crystals and compared with reference cards within a database that contains the diffraction patterns of known crystals. During a preliminary study of crystals harvested from various Cheddar cheese samples, we observed 2 slightly different but distinct diffraction patterns that suggested that calcium lactate pentahydrate may be present in 2 different crystalline forms. We hypothesized that the 2 diffraction patterns corresponded to 2 enantiomeric forms of calcium lactate pentahydrate (L- and DL-) that are believed to occur in Cheddar cheese, based on previous studies involving enzymatic analyses of the lactate enantiomers in crystals obtained from Cheddar cheeses. However, the powder X-ray diffraction database currently contains only one reference diffraction card under the title "calcium lactate pentahydrate." To resolve this apparent gap in the powder X-ray diffraction database, we generated diffraction patterns from reagent-grade calcium l-lactate pentahydrate and laboratory-synthesized calcium dl-lactate pentahydrate. From the resulting diffraction patterns we determined that the existing reference diffraction card corresponds to calcium dl-lactate pentahydrate and that the other form of calcium lactate pentahydrate observed in cheese crystals corresponds to calcium l-lactate pentahydrate. Therefore, this report presents detailed data from the 2 diffraction patterns, which may be used to prepare 2 reference diffraction cards that differentiate calcium l-lactate pentahydrate from calcium dl-lactate pentahydrate. Furthermore, we collected crystals from the exteriors and interiors of Cheddar cheeses to demonstrate the ability of powder X-ray diffraction to differentiate between the 2 forms of calcium lactate pentahydrate crystals in Cheddar cheeses. Powder X-ray diffraction results were validated using enzymatic assays for lactate enantiomers. These results demonstrated that powder X-ray diffraction can be used as a diagnostic tool to quickly identify different forms of calcium lactate pentahydrate that may occur in Cheddar cheese.

Surface roughness and packaging tightness affect calcium lactate crystallization on Cheddar cheese.

Calcium lactate crystals that sometimes form on Cheddar cheese surfaces are a significant expense to manufacturers. Researchers have identified several postmanufacture conditions such as storage temperature and packaging tightness that contribute to crystal formation. Anecdotal reports suggest that physical characteristics at the cheese surface, such as roughness, cracks, and irregularities, may also affect crystallization. The aim of this study was to evaluate the combined effects of surface roughness and packaging tightness on crystal formation in smoked Cheddar cheese. Four 20-mm-thick cross-section slices were cut perpendicular to the long axis of a retail block (~300g) of smoked Cheddar cheese using a wire cutting device. One cut surface of each slice was lightly etched with a cheese grater to create a rough, grooved surface; the opposite cut surface was left undisturbed (smooth). The 4 slices were vacuum packaged at 1, 10, 50, and 90kPa (very tight, moderately tight, loose, very loose, respectively) and stored at 1°C. Digital images were taken at 1, 4, and 8 wk following the first appearance of crystals. The area occupied by crystals and number of discrete crystal regions (DCR) were quantified by image analysis. The experiment was conducted in triplicate. Effects of storage time, packaging tightness, surface roughness, and their interactions were evaluated by repeated-measures ANOVA. Surface roughness, packaging tightness, storage time, and their 2-way interactions significantly affected crystal area and DCR number. Extremely heavy crystallization occurred on both rough and smooth surfaces when slices were packaged loosely or very loosely and on rough surfaces with moderately tight packaging. In contrast, the combination of rough surface plus very tight packaging resulted in dramatic decreases in crystal area and DCR number. The combination of smooth surface plus very tight packaging virtually eliminated crystal formation, presumably by eliminating available sites for nucleation. Cut-and-wrap operations may significantly influence the crystallization behavior of Cheddar cheeses that are saturated with respect to calcium lactate and thus predisposed to form crystals.

Effect of storage temperature on crystal formation rate and growth rate of calcium lactate crystals on smoked Cheddar cheeses.

Previous studies have shown that storage temperature influences the formation of calcium lactate crystals on vacuum-packaged Cheddar cheese surfaces. However, the mechanisms by which crystallization is modulated by storage temperature are not completely understood. The objectives of this study were to evaluate the effect of storage temperature on smoked Cheddar cheese surfaces for (1) the number of discrete visible crystals formed per unit of cheese surface area; (2) growth rate and shape of discrete crystals (as measured by area and circularity); (3) percentage of total cheese surface area occupied by crystals. Three vacuum-packaged, random weight (∼300 g) retail samples of naturally smoked Cheddar cheese, produced from the same vat of cheese, were obtained from a retail source. The samples were cut parallel to the longitudinal axis at a depth of 10mm from the 2 surfaces to give six 10-mm-thick slabs, 4 of which were randomly assigned to 4 different storage temperature treatments: 1, 5, 10°C, and weekly cycling between 1 and 10°C. Samples were stored for 30 wk. Following the onset of visible surface crystals, digital photographs of surfaces were taken every other week and evaluated by image analysis for number of discrete crystal regions and total surface area occupied by crystals. Specific discrete crystals were chosen and evaluated biweekly for radius, area, and circularity. The entire experiment was conducted in triplicate. The effects of cheese surface, storage temperature, and storage time on crystal number and total crystal area were evaluated by ANOVA, according to a repeated-measures design. The number of discrete crystal regions increased significantly during storage but at different rates for different temperature treatments. Total crystal area also increased significantly during storage, at rates that varied with temperature treatment. Storage temperature did not appear to have a major effect on the growth rates and shapes of the individual crystals that were chosen for analysis. The data indicated that the effect of storage temperature was complex, likely involving solubility changes, the formation of d(-) and l(+) lactic acid, and the occurrence of syneresis, which in turn affected the number of crystal formation sites and total crystal area on the cheese surface.

Chemical changes that predispose smoked Cheddar cheese to calcium lactate crystallization.

We have observed a high incidence of calcium lactate surface crystals on naturally smoked Cheddar cheese in the retail marketplace. The objective of this study was to identify chemical changes that may occur during natural smoking that render Cheddar cheese more susceptible to calcium lactate crystal formation. Nine random-weight (approximately 300 g) retail-packaged samples of smoked Cheddar cheese were obtained from a commercial manufacturer immediately after the samples were smoked for about 6 h at 20 degrees C in a commercial smokehouse. Three similarly sized samples that originated from the same 19.1-kg block of cheese and that were not smoked were also obtained. Within 2 d after smoking, 3 smoked and 3 control (not smoked) samples were sectioned into 5 subsamples at different depths representing 0 to 2, 2 to 4, 4 to 6, 6 to 8, and 8 to 10 mm from the cheese surface. Six additional smoked cheese samples were similarly sectioned at 4 wk and again at 10 wk of storage at 5 degrees C. Sample sections were analyzed for moisture, L(+) and D(-) lactate, pH, and water-soluble calcium. The effects of treatment (smoked, control), depth from cheese surface, and their interactions were analyzed by ANOVA according to a repeated measures design with 2 within-subject variables. Smoked samples contained significantly lower moisture and lower pH, and higher total lactate-in-moisture (TLIM) and water-soluble calcium-in-moisture (WSCIM) than control cheeses. Smoked samples also contained significant gradients of moisture, pH, TLIM, and WSCIM, with lower moisture and pH, and higher TLIM and WSCIM, occurring at the cheese surface. Gradients of moisture were still present in smoked samples at 4 and 10 wk of storage. In contrast, the pH, TLIM, and WSCIM equilibrated and showed no gradients at 4 and 10 wk. The results indicate that calcium and lactate in the serum phase of the cheese were elevated because of smoking, especially at the cheese surface immediately after smoking treatment, which presumably predisposes the smoked cheeses to increased susceptibility to calcium lactate surface crystallization.

Effect of pH on microstructure and characteristics of cream cheese.

This study evaluated the effect of pH on the microstructure of cream cheese and compared pH-induced changes in its microstructure with concomitant changes in cheese firmness and meltability. On 4 different days, experimental batches of cultured hot pack cream cheese were manufactured and analyzed for initial chemical composition. The cheeses were then sectioned into samples that were randomly assigned to 7 different treatment groups. Three groups were exposed to ammonia vapor for 1, 3, and 5 min to increase the pH; 3 groups were exposed to acetic acid vapor for 30, 60, and 90 min to decrease the pH; and 1 unexposed group served as the control. After equilibration at 4 degrees C, samples were analyzed for pH, firmness, meltability, and microstructure by scanning electron microscopy. The effects of experimental treatments on cheese pH, firmness, and meltability were analyzed by randomized complete block analysis of variance (ANOVA). Relationships between cheese pH and firmness and meltability were evaluated by regression. Experimental treatments significantly affected cheese pH, firmness, and meltability. Cheese firmness decreased and meltability increased with increasing pH from about pH 4.2 to 6.8. Cheese microstructure also changed dramatically over the same approximate pH range. Specifically, the volume of the protein network surrounding the fat droplets increased markedly with increasing pH, presumably due to casein swelling. These data support the hypothesis that protein-to-water interactions increased as the cheese pH increased, which gave rise to progressive swelling of the casein network, softer texture, and increased meltability.

Effect of pH on characteristics of low-moisture Mozzarella cheese during refrigerated storage.

This study evaluated the effect of cheese pH on proteolysis, calcium distribution, and functional characteristics of Mozzarella cheese. On 4 occasions, cultured low-moisture part-skim Mozzarella cheeses were obtained from a commercial producer on the day after manufacture. Cheese blocks were randomly assigned to 2 groups. One group was shredded, subdivided, and exposed to either ammonia vapor to increase the pH or HCl vapor to decrease the pH. Samples were vacuum packaged, stored at 4 degrees C, and analyzed for pH 4.6 and 12% TCA soluble nitrogen, apparent viscosity, free oil, and water-soluble calcium on days 5, 12, 22, and 40. The 2nd group was sectioned into 23-mm thick slabs and similarly exposed to either ammonia vapor to increase the pH or HCl vapor to decrease the pH. The slabs were vacuum packaged, stored at 4 degrees C, and analyzed for pH 4.6 and 12% TCA soluble nitrogen, TPA hardness, springiness and cohesiveness, and meltability on days 17, 29, and 41. Data were analyzed by ANOVA according to a spilt-plot design. Experimentally induced pH differences persisted and significantly affected TPA hardness, apparent viscosity, meltability, and water-soluble calcium throughout 40 d of storage, but did not affect soluble nitrogen changes. Thus, cheese pH affected functional characteristics and calcium distribution but did not affect proteolysis rates. Higher cheese pH resulted in a harder cheese that required longer aging to develop desirable melting characteristics, whereas cheese with lower pH developed desirable melting characteristics more quickly but had a shorter functional shelf life.

Characterization of calcium lactate crystals on cheddar cheese by image analysis.

Previous research demonstrated that crystal coverage on the surface of Cheddar cheese can be quantitatively and nondestructively measured using image analysis of digital photographs of the cheese surface. The objective of the present study was to extend image analysis methodology to quantify and characterize additional features of visible crystals on cheese surfaces as they grow over time. A random weight (approximately 300 g) retail sample of naturally smoked Cheddar cheese exhibiting white surface crystals was obtained from a commercial source. The total area occupied by crystals and total number of discrete crystal regions on one of the surfaces (approximately 55 x 120 mm) was measured at 3-wk intervals for 30 wk using image analysis. In addition, 5 small (approximately 0.3 mm radius) individual crystals on that surface were chosen for observation over the 30-wk period. The crystals were evaluated for area, radius, and shape factor (circularity) every third week using image analysis. The total area occupied by crystals increased in a linear manner (R(2) = 0.95) from about 0.44 to 7.42% of the total cheese surface area over the 30-wk period. The total number of discrete crystal regions also increased but in a nonlinear manner that was best described by a quadratic relationship. Measurement of discrete crystal regions underestimated the true number of crystals present at the cheese surface due to merging of adjacent crystals as they grew and merged into a single crystal region over time. Throughout this period, the shapes of the 5 individual crystals closely approximated perfect circles, except when adjacent crystals merged to form a single irregular crystal region, and the area occupied by each of the 5 crystals increased in a near-linear manner (R(2) = 0.95). Image analysis approaches may be used to evaluate crystal formation and growth rates and morphology on cheese.

Development and application of image analysis to quantify calcium lactate crystals on the surface of smoked Cheddar cheese.

Calcium lactate crystals that form white specks or haze on the surface of cheese constitute a significant quality problem for producers of Cheddar cheese. Subjective methods to evaluate crystal coverage of cheese surfaces have been reported previously, but objective methods are currently lacking. The objectives of this work were to develop and evaluate an objective method to measure the area occupied by calcium lactate crystals on surfaces of naturally smoked Cheddar cheese samples using digital photography and image analysis. Coefficients of variation ranged from 1.29 to 4.68% for 5 replicate analyses of 3 different cheese surfaces that ranged from approximately 2 to 49% of total surface area occupied by crystals. Thus, results showed a high degree of repeatability for the 3 cheese surfaces, which ranged from very slight and geometrically simple to very heavy and geometrically complex crystal coverage. The method underestimated total area occupied by crystals on the 3 surfaces by 0.24 to 4.83% unless the fainter crystal regions that went undetected during initial thresholding were manually segmented and quantified. The wet weight of crystal substance collected per unit of surface area from 20 different cheese samples increased exponentially as the percentage of total surface area occupied by crystals increased. These data were consistent with subjective observations that crystal regions appeared to grow vertically as well as horizontally as they expanded to occupy greater surface area. Image analysis was well suited for evaluating changes in crystal coverage during cheese aging because measurements were made nondestructively and with minimal disruption to the cheese. The area occupied by crystals on 6 different surfaces from 3 different cheese samples increased linearly (R2 = 0.94 to 0.99) during storage at 4 degrees C for up to 33 wk. However, the rates of increase differed significantly among the 3 cheese samples. Image analysis may serve as a useful tool to quantitatively evaluate the effects of factors such as cheese composition, packaging conditions and storage temperature on rate of crystal growth and time of crystal appearance during storage.

Compositional factors associated with calcium lactate crystallization in smoked Cheddar cheese.

Previous researchers have observed that surface crystals of calcium lactate sometimes develop on some Cheddar cheese samples but not on other samples produced from the same vat of milk. The causes of within-vat variation in crystallization behavior have not been identified. This study compared the compositions of naturally smoked Cheddar cheese samples that contained surface crystals with those of samples originating from the same vat that were crystal-free. Six pairs of retail samples (crystallized and noncrystallized) produced at the same cheese plant on different days were obtained from a commercial source. Cheese samples were 5 to 6 mo old at the time of collection. They were then stored for an additional 5 to 13 mo at 4 degrees C to ensure that the noncrystallized samples remained crystal-free. Then, the crystalline material was removed and collected from the surfaces of crystallized samples, weighed, and analyzed for total lactic acid, L(+) and D(-) lactic acid, Ca, P, NaCl, moisture, and crude protein. Crystallized and noncrystallized samples were then sectioned into 3 concentric subsamples (0 to 5 mm, 6 to 10 mm, and greater than 10 mm depth from the surface) and analyzed for moisture, NaCl, titratable acidity, L(+) and D(-) lactic acid, pH, and total and water-soluble calcium. The data were analyzed by ANOVA according to a repeated measures design with 2 within-subjects variables. The crystalline material contained 52.1% lactate, 8.1% Ca, 0.17% P, 28.5% water, and 8.9% crude protein on average. Both crystallized and noncrystallized cheese samples contained significant gradients of decreasing moisture from center to surface. Compared with noncrystallized samples, crystallized samples possessed significantly higher moisture, titratable acidity, L(+) lactate, and water soluble calcium, and significantly lower pH and NaCl content. The data suggest that formation of calcium lactate crystals may have been influenced by within-vat variation in salting efficacy in the following manner. Lower salt uptake by some of the cheese curd during salting may have created pockets of higher moisture and thus higher lactose within the final cheese. When cut into retail-sized chunks, the lower salt, higher moisture samples contained more lactic acid and thus lower cheese pH, which shifted calcium from the insoluble to the soluble state. Lactate and soluble calcium contents in these samples became further elevated at the cheese surface because of dehydration during smoking, possibly triggering the formation of calcium lactate crystals.

Solubility and relative absorption of copper, iron, and zinc in two milk-based liquid infant formulae.

The relative solubility of copper, iron, and zinc in two experimental liquid infant formulae are examined. The results of these trials suggest that substituting organic forms of copper and iron in the mix results in an almost three-fold increase in their solubility (iron lactate, 73.4% vs ferrous sulfate, 27.6% and copper gluconate, 11.3% vs cupric sulfate, 3.0%). Organic zinc substitutes did not show this pattern of increased solubility Electron microscopy was employed to document the changes in protein-protein and protein-lipid interactions and to examine the pattern of electron-dense precipitates in the two experimental formulae. Electron micrographs of the liquid infant formula that had been formulated using inorganic salts (sulfates) showed extensive attachment of denatured whey proteins and casein micelles to the oil droplet surfaces and the surface of the oil droplets were also punctuated by electron-dense granule. The oil droplets of the formula produced using organic versions of the mineral salts were smooth and clear of electron-dense deposits. Experiments were designed to determine whether the observed changes in solubility and microstructure were correlated with increases in relative absorption of the minerals. We applied a technique of in vitro acidification followed by a peptic digestion of the two experimental infant formulae with human milk samples as controls. Coupling this in vitro digestion with an absorption model consisting of live isolated intestinal loops from guinea pig we were able to assess the relative absorption of copper, iron and zinc in the test digests. The relative absorption of the three minerals from digests of human milk was significantly higher than for either of the experimental formulae. Relative mineral absorption from digests of the two experimental infant formulae tested was only significantly different (P < 0.05) for Fe. Based on the results from this study we can conclude that substituting organic forms of iron in bovine milk-base infant formulae would have beneficial effects on both the solubility and bioavailability of this important micronutrient.

Effect of milk preacidification on low fat mozzarella cheese: II. Chemical and functional properties during storage.

The effect of milk preacidification on cheese manufacturing, chemical properties, and functional properties of low fat Mozzarella cheese was determined. Four vats of cheese were made in 1 d using no preacidification (control), preacidification to pH 6.0 and pH 5.8 with acetic acid, and preacidification to pH 5.8 with citric acid. This process was replicated four times. Modifications in the typical Mozzarella manufacturing procedures were necessary to accommodate milk preacidification. The chemical composition of the cheeses was similar among the treatments, except the calcium content and calcium as a percentage of protein were lower in the preacidified treatments. During refrigerated storage, the chemical and functional properties of low fat Mozzarella were affected the most by milk preacidification to pH 5.8 with citric acid. The amount of expressible serum, unmelted cheese whiteness, initial unmelted hardness, and initial apparent viscosity were lower with preacidification. The reduction in initial unmelted cheese hardness and initial apparent viscosity in the pH 5.8 citric treatments represents an improvement in the quality of low fat Mozzarella cheese that allows the cheese to have better pizza bake characteristics with shorter time of refrigerated storage.

Effect of milk preacidification on low fat mozzarella cheese: III. Post-melt chewiness and whiteness.

The effect of calcium reduction (as a result of milk preacidification) on post-melt chewiness and whiteness of low fat Mozzarella cheese was determined. Four vats (230 kg of milk per vat) of cheese were made in 1 d using no preacidification (control), preacidification pH 6.0 and pH 5.8 with acetic acid, and preacidification to pH 5.8 with citric acid. Cheese manufacture was repeated on four different days using a randomized complete block design. The total calcium content and the water-insoluble calcium content of the cheese were lower in the cheeses made from preacidified milks. The amount of water-soluble and water-insoluble calcium changed during refrigerated storage, as did pH. The post-melt chewiness and whiteness of low fat Mozzarella cheese were affected by milk preacidification. The largest level of calcium reduction and modification in post-melt chewiness and whiteness occurred in the pH 5.8 citric treatment. Multiple regression analysis of post-melt chewiness and cheese whiteness at 38 degrees C after heating and cooling indicated that both water-insoluble calcium and proteolysis were strongly associated with changes in the post-melt chewiness and whiteness of low fat Mozzarella cheese. High levels of proteolysis and low levels of water-insoluble calcium were associated with decreased post-melt chewiness and whiteness of low fat Mozzarella cheese.

Moisture variations in brine-salted pasta filata cheese.

A study was made of the moisture distribution in brine-salted pasta filata cheese. Brine-salted cheeses usually develop reasonably smooth and predictable gradients of decreasing moisture from center to surface, resulting from outward diffusion of moisture in response to inward diffusion of salt. However, patterns of moisture variation within brine-salted pasta filata cheeses, notably pizza cheese, are more variable and less predictable because of the peculiar conditions that occur when warm cheese is immersed in cold brine. In this study, cold brining resulted in less moisture loss from the cheese surface to the brine. Also it created substantial temperature gradients within the cheese, which persisted after brining and influenced the movement of moisture within the cheese independently of that caused by the inward diffusion of salt. Depending on brining conditions and age, pizza cheese may contain decreasing, increasing, or irregular gradients of moisture from center to surface, which may vary considerably at different locations within a single block. This complicates efforts to obtain representative samples for moisture and composition testing. Dicing the entire block into small (e.g., 1.5 cm) cubes and collecting a composite sample after thorough mixing may serve as a practical sampling approach for manufacturers and users of pizza cheese that have ready access to dicing equipment.

Effect of milk preacidification on low fat mozzarella cheese. I. Composition and yield.

The effect of milk preacidification with acetic or citric acid on the composition and yield of low fat Mozzarella cheese was determined. Two cheese manufacturing trials were conducted. In trial 1, three vats (230 kg of milk per vat) of cheese were made in 1 d using no preacidification (control) and preacidification to pH 6.0 and pH 5.8 with citric acid. In trial 2, four vats (230 kg of milk per vat) of cheese were made in 1 d using no preacidification (control), preacidification to pH 6.0 and 5.8 with acetic acid, and preacidification to pH 5.8 with citric acid. Cheese manufacture was repeated on three different days in trial 1 and four different days in trial 2 using a randomized-complete block design. Preacidification to pH 5.8 with citric acid decreased cheese calcium more than preacidification to pH 5.8 with acetic acid. Preacidification with citric acid in trial 1 decreased protein recovery in the cheese, and there was a trend for decreased protein recovery in the cheese for trial 2. Differences in fat recovery due to preacidification varied, sometimes being lower than the control other times being higher than the control. The reduction in calcium and protein recovery in the cheese caused by preacidification lowered composition adjusted cheese yield and yield efficiency. Yield efficiency was reduced by about 2.5 and 5.5%, respectively, with preacidification to pH 6.0 and 5.8 with citric acid. Yield efficiency was reduced by about 2.2 and 3.4%, respectively, with preacidification to pH 6.0 and pH 5.8 with acetic acid.

Whiteness change during heating and cooling of Mozzarella cheese.

Whiteness (L-value) changes in low-fat and low-moisture, part-skim Mozzarella cheeses during heating (7 to 60 degrees C) and cooling (60 to 7 degrees C) were evaluated. In low-fat Mozzarella, a large increase in whiteness was observed during heating, and a decrease in whiteness was observed during cooling. In low-moisture, part-skim Mozzarella, the whiteness changes during heating and cooling were smaller. Serum phase was removed from low-fat and low-moisture, part-skim Mozzarella cheeses. White protein gels were formed when the isolated serum phase from either low-fat or low-moisture, part-skim Mozzarella was heated. The white gel that formed was composed predominantly of casein and casein proteolysis products. The gel might have been produced by heat-induced, hydrophobic protein-protein interactions, and it tended to dissociate when cooled. Formation of a gel during heating increased light scattering, which increased the L-value. The gel dissociated during cooling and no longer scattered light, which decreased the L-value. We hypothesized that a gel, which was reversible, formed in the serum phase of cheese during heating and might have been responsible for the observed changes in the L-value of low-fat Mozzarella cheese during heating and cooling. The additional fat in low-moisture, part-skim Mozzarella compared with low-fat Mozzarella masked some of the color changes in the serum phase of low-moisture, part-skim Mozzarella. A model was developed to describe the contributions of the casein matrix plus serum phase of Mozzarella cheese and the contribution of fat to the changes in whiteness of Mozzarella cheese during heating and cooling.

Effect of fat reduction on chemical composition, proteolysis, functionality, and yield of Mozzarella cheese.

Mozzarella cheese was made from skim milk standardized with cream (unhomogenized, 40% milk fat) to achieve four different target fat percentages in the cheese (ca. 5, 10, 15, and 25%). No statistically significant differences were detected for cheese manufacturing time, stretching time, concentration of salt in the moisture phase, pH, or calcium as a percentage of the protein in the cheese between treatments. As the fat percentage was reduced, there was an increase in the moisture and protein content of the cheese. However, because the moisture did not replace the fat on an equal basis, there was a significant decrease in the moisture in the nonfat substance in the cheese as the fat percentage was reduced. This decrease in total filler volume (fat plus moisture) was associated with an increase in the hardness of the unmelted cheese. Whiteness and opacity of the unmelted cheese decreased as the fat content decreased. Pizza baking performance, meltability, and free oil release significantly decreased as the fat percentage decreased. The minimum amount of free oil release necessary to obtain proper functionality during pizza baking was between 0.22 and 2.52 g of fat/100 g of cheese. Actual cheese yield was about 30% lower for cheese containing 5% fat than for cheese with 25% fat. Maximizing fat recovery in the cheese becomes less important to maintain high cheese yield, and moisture control and the retention of solids in the water phase become more important as the fat content of the cheese is reduced.

Susceptibility of beta-lactoglobulin and sodium caseinate to proteolysis by pepsin and trypsin.

The susceptibility of beta-LG and sodium caseinate to proteolysis by pepsin and trypsin was investigated using SDS or urea-PAGE. The effects were studied of heat, urea, and 2-mercaptoethanol on proteolysis. Native beta-LG was resistant to hydrolysis by pepsin or trypsin because of its compact globular structure. Heat treatment of beta-LG solutions at 90 to 100 degrees C for 5 or 10 min caused changes in the structure or conformation of the protein that rendered it accessible to pepsin and enhanced the extent of proteolysis by trypsin. The susceptibility of beta-LG to proteolysis by pepsin was markedly increased in the presence of urea (3 to 6 M), and the effect was reversible after removal of urea by dialysis. Proteolysis by trypsin was also increased by the presence of 2% 2-mercaptoethanol. Sodium caseinate was very accessible to pepsin without pretreatment and was extensively hydrolyzed at pH 1 to 5 in the presence of 5 M urea (which prevented the protein from precipitation in the isoelectric region); optimal pH was about 2. The activity of pepsin on sodium caseinate at pH 2 was not significantly affected by urea concentration up to about 8 M. The results indicated that the changes in conformation and structure of beta-LG that were induced by heating, reduction, or urea rendered the protein susceptible to peptic hydrolysis.