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PubMed2001

Recombinant lamb chymosin as an alternative coagulating enzyme in cheese production.

Recombinant lamb chymosin (RLC) was prepared and tested for its potential use in cheese production. The milk clotting activity and proteolytic activity of RLC were evaluated in comparison with commercial recombinant calf chymosin (RCC), cow rennet (CR), and microbial coagulant (MC). RLC, RCC, and MC showed similar responses to pH, with a sharp increase of the coagulation time at pH 6.6 to 6.8 and decrease of curd firmness at the pH 6.5 to 6.6. In the case of CR, we observed two clear increases in the coagulation time and decreases in the curd firmness, at pH 6.4 to 6.5 and 6.6 to 6.8. Optimal clotting activity was obtained for RLC at 40 degrees C, for both CR and RCC at 45 degrees C, and for MC at 60 degrees C. The temperature instability of RLC at temperatures above 45 degrees C could constitute a benefit in making hard cheese varieties. The additon of CaCl2 to milk resulted in enhanced clotting activity of all coagulants, most prominently for CR. The proteolytic activity of RLC was significantly lower from that of CR but not significantly different from the activity of RCC. The lower proteolytic activity in the cheese made with RLC did not have negative effect on organoleptic properties. The overall quality of the cheese made with RLC was at least comparable to that of the cheese made with RCC, and both cheeses were better scored than the cheese made with CR.

Animals↗
PubMed2002

Effect of pH and calcium concentration on proteolysis in mozzarella cheese.

Low-moisture Mozzarella cheeses (LMMC), varying in calcium content and pH, were made using a starter culture (control; CL) or direct acidification (DA) with lactic acid or lactic acid and glucono-delta-lactone. The pH and calcium concentration significantly affected the type and extent of proteolysis in Mozzarella cheese during the 70-d storage period at 4 degrees C. For cheeses with a similar pH, reducing the calcium-to-casein ratio from -29 to 22 mg/g of protein resulted in marked increases in moisture content and in primary and secondary proteolysis, as indicated by polyacrylamide gel electrophoresis and higher levels of pH 4.6- and 5%-PTA-soluble N. Increasing the pH of DA cheeses of similar moisture content, from approximately 5.5 to 5.9, while maintaining the calcium-to-casein ratio almost constant at approximately 29 mg/g, resulted in a decrease in primary proteolysis but had no effect on secondary proteolysis. Comparison of CL and DA cheeses with a similar composition showed that the CL cheese had higher levels of alpha(s1)-CN degradation, pH 4.6- and 5%-PTA-soluble N. Analysis of pH 4.6-soluble N extracts by reverse-phase HPLC showed that the CL cheese had higher concentrations of compounds with low retention times, suggesting higher concentrations of low molecular mass peptides and free amino acids.

Animals↗
PubMed2002

Temperature-induced moisture migration in reduced-fat Cheddar cheese.

Moisture migration during cooling of 290-kg Cheddar cheese blocks is a problem. The problem is of greater magnitude in reduced and low fat varieties. The objective of this study was to design and evaluate the performance of a laboratory-scale apparatus for simulation of temperature induced moisture migration in 290-kg blocks of Cheddar cheese. Two apparati were designed to produce a systematic temperature gradient in small cheese slabs over a 36-h period to simulate the temperature gradient that develops during cooling of a 290-kg block. One of the apparati was designed to induce a moisture migration downwards with gravity and the other against gravity. The apparati produced moisture migration ranges of 9.7 and 6.4%, for the apparatus to induce moisture migration downwards and upwards, respectively. The moisture moved from areas of warm cheese to areas of cold cheese during cooling, as occurs in 290-kg blocks. These ranges were comparable to those obtained with 290-kg reduced-fat Cheddar blocks. In addition, small but significant differences in pH were created within slabs. The direct effect of the temperature gradient on moisture migration within cheese slabs appeared to be more important than the possible impact of the small pH gradient produced within the cheese by the temperature gradient.

Cheese↗
PubMed2002

Influence of starter and nonstarter on the formation of biogenic amine in goat cheese during ripening.

Two commercial starters were investigated for their potential ability to decarboxylate amino acids during goat cheese ripening. Two batches of goat cheese were produced with identical pasteurized milk but different starter cultures. One of them contained Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris and the other Lactococcus lactis subsp. lactis. The amine contents, microbial counts, proteolysis-related parameters, pH, total solids and salt content were studied in raw materials and cheeses. In raw materials, polyamines were the prevailing amines, whereas the main amines in cheeses were putrescine, tryptamine and, in particular, tyramine (94.59 mg/kg). Aerobic mesophilic microorganisms and Lactococcus counts increased throughout ripening, while Enterobacteriaceae were no longer detectable in cheese after 30 days of ripening. Amine concentration rose during cheese ripening in both batches. Moreover, the decarboxylase activity of microorganisms isolated from samples during cheese ripening was assayed and discussed.

Animals↗
PubMed2002

Estimation and fortification of vitamin D3 in pasteurized process cheese.

The objective of this study was to develop methods for the estimation and fortification of vitamin D3 in pasteurized Process cheese. Vitamin D3 was estimated using alkaline saponification at 70 degrees C for 30 min, followed by extraction with petroleum ether:diethyl ether (90:10 vol/vol) and HPLC. The retention time for vitamin D3 was approximately 9 min. A standard curve with a correlation coefficient of 0.972 was prepared for quantification of vitamin D3 in unknown samples. In the second phase of the study, pasteurized Process cheeses fortified with commercial water- or fat-dispersible forms of vitamin D3 at a level of 100 IU per serving (28 g) were manufactured. There was no loss of vitamin D3 during Process cheese manufacture, and the vitamin was uniformly distributed. No losses of the vitamin occurred during storage of the fortified cheeses over a 9-mo period at 21 to 29 degrees C and 4 to 6 degrees C. There was an approximately 25 to 30% loss of the vitamin when cheeses were heated for 5 min in an oven maintained at 232 degrees C. Added vitamin D3 did not impart any off flavors to the Process cheeses as determined by sensory analysis. There were no differences between the water- and fat-dispersible forms of the vitamin in the parameters measured in fortified cheeses.

Cheese↗
PubMed2003

Proteolysis, volatile compounds, and sensory evaluation in Hispánico cheese manufactured with the addition of a thermophilic adjunct culture, nisin, and calcium alginate-nisin microparticles.

Nisin, free or incorporated in calcium alginate microparticles, was added to pasteurized milk (80% cows' and 20% ewes' milk) used for the manufacture of Hispánico cheese with a mesophilic starter and a thermophilic adjunct culture of high aminopeptidase activity. Addition of nisin incorporated in microparticles promoted early lysis of thermophilic adjunct culture bacteria. Extracellular aminopeptidase activity in 1-d-old cheese made with both thermophilic adjunct culture and nisin incorporated in microparticles was 1.8-fold higher than in cheese made with thermophilic adjunct culture and free nisin and 2.0-fold higher than in cheese made only with thermophilic adjunct culture without any addition of nisin. Addition of nisin, free or incorporated in microparticles, did not influence cheese proteolysis measured using hydrophilic or hydrophobic peptides, o-phthaldialdehyde ripening index, or free amino acids. Moreover, a total of 37 volatile compounds were identified in the volatile fraction of Hispánico cheese, using a dynamic headspace technique with a purge and trap system followed by a gas chromatography mass spectrometry analysis. The volatile compound profile was not influenced by nisin addition, either free or incorporated in microparticles, but addition of thermophilic adjunct culture enhanced the formation of 2-butanone, diacetyl, 2,3-pentanedione and acetoin and improved the flavor quality (sensory panel) of cheese.

Alginates↗
PubMed2004

Molecular Subtyping and Tracking of Listeria monocytogenes in Latin-Style Fresh-Cheese Processing Plants.

Latin-style fresh cheeses, which have been linked to at least 2 human listeriosis outbreaks in the United States, are considered to be high-risk foods for Listeria monocytogenes contamination. We evaluated L. monocytogenes contamination patterns in 3 Latin-style fresh-cheese processing plants to gain a better understanding of L. monocytogenes contamination sources in the manufacture of these cheeses. Over a 6-mo period, 246 environmental samples were collected and analyzed for L. monocytogenes using both the Food and Drug Administration (FDA) method and the Biosynth L. monocytogenes detection system (LMDS). Finished cheese samples from the same plants (n = 111) were also analyzed by the FDA method, which was modified to include L. monocytogenes plating medium (LMPM) and the L. monocytogenes confirmatory plating medium (LMCM) used in the LMDS method. Listeria monocytogenes was detected in 6.3% of cheese and 11.0% of environmental samples. Crates, drains, and floor samples showed the highest contamination rates, with 55.6, 30.0, and 20.6% L. monocytogenes positive samples, respectively. Finished products and food contact surfaces were positive in only one plant. The FDA method showed a higher sensitivity than the LMDS method for detection of L. monocytogenes from environmental samples. The addition of LMPM and LMCM media did not further enhance the performance of the FDA method for L. monocytogenes detection from finished products. Molecular subtyping (PCR-based allelic analysis of the virulence genes actA and hly and automated ribotyping) was used to track contamination patterns. Ribotype DUP-1044A, which had previously been linked to a 1998 multistate human listeriosis outbreak in the United States, was the most commonly identified subtype (20/36 isolates) and was isolated from 2 plants. This ribotype was persistent and widespread in one factory, where it was also responsible for the contamination of finished products. We hypothesize that this ribotype may represent a clonal group with a specific ability to persist in food processing environments. While previous listeriosis outbreaks were linked to Latin-style fresh cheeses made from unpasteurized milk, the presence of this organism in pasteurized cheese products illustrates that persistent environmental contamination also represents an important source of finished product contamination.

Cheese↗
PubMed2004

Deacidification by Debaryomyces hansenii of smear soft cheeses ripened under controlled conditions: relative humidity and temperature influences.

Model smear soft cheeses were prepared from pasteurized milk inoculated with Debaryomyces hansenii (304, GMPA) and Brevibacterium aurantiacum (ATCC 9175) under aseptic conditions. Debaryomyces hansenii growth and curd deacidification were studied in relation to ripening chamber temperature and relative humidity (RH). A total of 9 descriptors, mainly based on kinetic data, were defined to represent D. hansenii growth (2 descriptors), cheese deacidification (5 descriptors), and cheese ripening (2 descriptors). Regardless of the temperature, when the RH was 85%, D. hansenii growth was inhibited due to limitation of carbon substrate diffusions; consequently, cheese deacidification did not take place. Debaryomyces hansenii growth was most prolific when the temperature was 16 degrees C, and the RH was 95%. Kinetic descriptors of lactate consumption and pH increase were maximal at 16 degrees C and 100% RH. Under these 2 ripening conditions, on d 14 (packaging) the creamy underrind represented a third of the cheese; however, at the end of ripening (d 42), cheese was too liquid to be sold. Statistical analysis showed that the best ripening conditions to achieve an optimum between deacidification and appearance of cheeses (thickness of the creamy underrind) were 12 degrees C and 95 +/- 1% RH.

Brevibacterium↗
PubMed2005

Utilization of front-face fluorescence spectroscopy for analysis of process cheese functionality.

The purpose of this study was to evaluate the feasibility of front-face fluorescence spectroscopy (FFFS) to predict the meltability of process cheese spreads or products. Twenty-seven commercial samples from 3 manufacturers were used in this study. Each sample was analyzed using dynamic stress rheometry, which was used to calculate the meltability index (temperature at tandelta = 1). Additionally, fluorescence spectra of tryptophan (excitation: 290 nm; emission: 305 to 400 nm) were collected on each sample at 20 degrees C using a front-face accessory. Fluorescence spectrum for each sample consisted of an average of 36 scans (6 scans performed on 6 replicates). The spectral data set consisted of normalized and mean-centered spectra from all the samples. Multivariate statistical analysis was used to correlate spectral data with cheese meltability index as measured by dynamic stress rheometry. A prediction model was developed using partial least square regression and was calibrated using a cross-validation method. A correlation coefficient of 0.93 was obtained between fluorescence spectra and cheese meltability. The regions 335 to 350 nm and 385 to 400 nm had the highest correlation to cheese meltability. A negative correlation between the peak height of tryptophan (335 to 350 nm) and cheese meltability index was observed. This correlation may be due to presence of tryptophan residues in a more hydrophobic environment in stronger emulsions as compared with a more polar environment in weak emulsions. These results indicate that the melt properties of process cheese spreads or products are related to molecular structure that can be measured using FFFS. Hence, FFFS can be used as an analysis technique to predict process cheese meltability.

Cheese↗
PubMed2005

Consumer acceptability of conjugated linoleic acid-enriched milk and cheddar cheese from cows grazing on pasture.

Two experiments were conducted to study the consumer acceptability attributes of conjugated linoleic acid (CLA)-enriched milk and cheese from cows grazing on pasture. In experiment 1, 15 cows were fed either a diet containing 51% alfalfa hay plus corn silage and 49% concentrate [total mixed ration (TMR)], were grazed on pasture, or were grazed on pasture and received 3.2 kg/d of a grain mix. The grain mix contained 75% full-fat extruded soybeans (FFES), 10% corn, 10% beet pulp, and 5% molasses. During the final 3 wk of the 6-wk experiment, milk was evaluated for sensory attributes. In experiment 2, 18 cows were fed similar diets as in experiment 1, except replacing the group of cows grazed on pasture and receiving the grain mix was a group of cows grazed on pasture and receiving 2.5 kg/d per cow of the FFES; Cheddar cheese was manufactured from milk. Average CLA contents (g/100 g of fatty acid methyl esters) were 0.52, 1.63, and 1.69 in milk and 0.47, 1.47, and 1.46 in cheese from cows fed a TMR, grazed on pasture, and grazed on pasture and fed the grain mix, respectively. An open and trained panel evaluated CLA-enriched milk for mouth-feel, color, flavor, and quality and evaluated cheese for color, flavor, texture, and quality. Open and trained panel evaluations of milk and cheese showed no differences among treatments for any of the attributes, except that the trained panel detected a more barny flavor in milk from cows grazing pasture compared with milk from cows fed the TMR only. Results suggest that consumer acceptability attributes of CLA-enriched milk and cheese from cows grazing pasture is similar to those of milk and cheese with low levels of CLA.

Animals↗
PubMed2005

Traceability of Asiago mountain cheese: a rapid, low-cost analytical procedure for its identification based on solid-phase microextraction.

The traceability of Asiago mountain cheese was established by analyzing samples of herbaceous species, milk, and cheese of mountain origin using the head-space solid-phase microextraction sampling procedure coupled with gas chromatography-mass spectrometry. As preliminary work had highlighted the characteristic presence of sesquiterpenes in Asiago mountain cheese, these species were considered effective markers of mountain origin. Systematic qualitative analysis, carried out using a carboxen/polydimethylsiloxane fiber, revealed several sesquiterpenes in mountain herbage and milk, in particular beta-caryophyllene and alpha-humulene, in Asiago mountain cheese, confirming sesquiterpenes as markers of cheese produced from animals grazing on mountain pastures. Analysis was performed on 19 samples of herbage, 8 of milk, and 8 of cheese, collected in summer from 4 mountain farms on the Asiago plateau. For quantitative analysis of caryophyllene in cheese, polydimethylsiloxane fiber sampling, coupled with the standard addition method to eliminate matrix effect, was preferred. The amount of beta-caryophyllene found ranged from 21 to 65 microg/kg.

Altitude↗
PubMed2005

Response of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella typhimurium, and Staphylococcus aureus to the thermal stress occurring in model manufactures of Grana Padano cheese.

The purpose of this research was to investigate the effect of temperature in the technology of production of Grana cheese against Escherichia coli O157:H7, Listeria monocytogenes, Salmonella Typhimurium, and Staphylococcus aureus. According to the technology of production, the cheese curds are cooked at 55 degrees C and then cooled at room temperature (25 degrees C). A curd-cooling model was developed to estimate the temperature variation across the curd during cooling, and the thermal stress was applied to the pathogens according to the model in model-scale productions of Grana cheese artificially contaminated with approximately 10(4) cfu/mL of the selected pathogens. According to the numerical results, the initial temperature inside the cheese is kept at almost the initial value (above 50 degrees C) for at least 4 h during cooling, whereas the crust of the curd cools rapidly to 30 degrees C in the first hour. The best case was that of the core of the cheese where the high temperature was able to efficiently eliminate the contaminating pathogens. Moreover, the worst case was where the external ring of the curd in which a more rapid cooling allowed bacterial survival. Therefore, the thermal stress in the technology of production of Grana cheese can be only partially effective in the control of the selected pathogens. However, the whole technology of production includes other hurdles that can affect the survival of the pathogens and that need to be taken into account as a whole to evaluate the safety of Grana Padano cheese.

Cheese↗
PubMed2006

Effect of milk protein standardization using different methods on the composition and yields of Cheddar cheese.

Two sets of Cheddar cheese were made in which the milk protein level (%, wt/wt) was increased from 3.3 (Control A, CA) to 3.6 (set A) or from 3.3 (control B, CB) to 4.0 (set B) by the addition of phosphocasein (PC), milk protein concentrate (MPC), or freshly prepared ultrafiltered milk retentate (UFR). The cheeses were denoted CA, PCA, MPCA, and UFRA from set A, and CB, PCB, MPCB, and UFRB, from set B, respectively. The level of cheese moisture decreased significantly on increasing milk protein level from 3.3 to 3.6 or 4.0% (wt/wt), but was not affected significantly by the method of protein standardization. The percentage milk fat recovered to cheese increased significantly on increasing the level of milk protein from 3.3 to 3.6% (wt/wt) with PC, and from 3.3 to 4.0% (wt/wt) with PC, MPC, and UFR. Increasing milk protein level from 3.3 to 4.0% (wt/wt) with PC significantly increased the percentage of milk protein recovered to cheese. Actual cheese yield increased significantly with milk protein level. The yield of cheese per 100 kg of milk normalized to reference levels of fat (3.4%, wt/wt) and casein (2.53%, wt/wt) indicated no significant effects of protein content or standardization treatment on yield. However, the moisture-adjusted yield per 100 kg of milk with reference levels of fat and casein increased significantly on increasing the protein content from 3.3 to 3.6% (wt/wt) with MPC and from 3.3 to 4.0% (wt/wt) with PC, MPC, and UFR.

Animals↗
PubMed2006

Proteolysis of Hispanico cheese manufactured using lacticin 481-producing Lactococcus lactis ssp. lactis INIA 639.

Hispánico cheese was manufactured using lacticin 481-producing Lactococcus lactis ssp. lactis INIA 639, bacteriocin-nonproducing L. lactis ssp. lactis INIA 437, or a combination of both strains, as starter cultures. Lactobacillus helveticus LH 92, a culture of high amino-peptidase activity sensitive to lacticin 481, was added to all vats. Milk inoculation with the bacteriocin producer promoted early lysis of Lb. helveticus cells in cheese. Cell-free aminopeptidase activity in cheese made with the 3 lactic cultures was 1.8 times the level reached in cheese made only with L. lactis strain INIA 437 and Lb. helveticus, after 15 d of ripening. Proteolysis (as estimated by the o-phthaldialdehyde method) in cheese made with the 3 lactic cultures was twice as high, and the level of total free amino acids 2.4 times the level found in cheese made only with L. lactis strain INIA 437 and Lb. helveticus, after 25 d of ripening. Hydrophobic and hydrophilic peptides and their ratio were at the lowest levels in cheese made with the 3 lactic cultures, which received the lowest scores for bitterness and the highest scores for taste quality.

Amino Acids↗
PubMed2006

Effect of protein composition on the cheese-making properties of milk from individual dairy cows.

The objective of this study was to evaluate the effect of variations in milk protein composition on milk clotting properties and cheese yield. Milk was collected from 134 dairy cows of Swedish Red and White, Swedish Holstein, and Danish Holstein-Friesian breed at 3 sampling occasions. Concentrations of alphaS1-, beta-, and kappa-casein (CN), alpha-lactalbumin, and beta-lactoglobulin (LG) A and B were determined by reversed phase liquid chromatography. Cows of Swedish breeds were genotyped for genetic variants of beta- and kappa-CN. Model cheeses were produced from individual skimmed milk samples and the milk clotting properties were evaluated. More than 30% of the samples were poorly coagulating or noncoagulating, resulting in weak or no coagulum, respectively. Poorly and noncoagulating samples were associated with a low concentration of kappa-CN and a low proportion of kappa-CN in relation to total CN analyzed. Furthermore, the kappa-CN concentration was higher in milk from cows with the AB genotype than the AA genotype of kappa-CN. The concentrations of alphaS1-, beta-, and kappa-CN and of beta-LG B were found to be significant for the cheese yield, expressed as grams of cheese per one hundred grams of milk. The ratio of CN to total protein analyzed and the beta-LG B concentration positively affected cheese yield, expressed as grams of dry cheese solids per one hundred grams of milk protein, whereas beta-LG A had a negative effect. Cheese-making properties could be improved by selecting milk with high concentrations of alphaS1-, beta-, and kappa-CN, with high kappa-CN in relation to total CN and milk that contains beta-LG B.

Animals↗
PubMed2006

Use of the acoustic impulse-response technique for the nondestructive assessment of Manchego cheese texture.

Manchego cheese pieces were hit with an impact probe and the acoustic response was recorded, analyzed, and used to assess the textural characteristics of the cheese pieces. The textural parameters measured by traditional instrumental methods increased during ripening, although the pattern of the increase was different for different batches. For the 2 acoustic impact probes used in this study, a change in the frequency spectrum took place as cheese matured, increasing higher frequencies and the energy content. Multiple linear regression (MLR) and partial least square regression (PLSR), considering the acoustical variables extracted from the spectrum, allowed for a good estimation of cheese texture. The textural characteristics of the cheese surface and in particular the maximum force in compression experiments (R(2) > 0.937 for MLR and R(2) > 0.852 for PLSR) were accurately predicted by the acoustic method; however, the texture of the central layers of the cheese are poorly assessed (R(2) < 0.720). The results obtained show the feasibility of using acoustic systems to assess Manchego cheese texture, aiding its classification.

Acoustics↗
PubMed2006

Effect of high-pressure treatment and a bacteriocin-producing lactic culture on the proteolysis, texture, and taste of Hispánico cheese.

The effects of high-pressure treatment, by itself or in combination with a bacteriocin-producing culture added to milk, on the proteolysis, texture, and taste of Hispánico cheese were investigated. Two vats of cheese were manufactured from a mixture of cow and ewe milk. Milk in one vat was inoculated with 0.5% Lactococcus lactis ssp. lactis INIA 415, a nisin Z and lacticin 481 producer; 0.5% L. lactis ssp. lactis INIA 415-2, a bacteriocin-nonproducing mutant; and 2% of a commercial Streptococcus thermophilus culture. Milk in the other vat was inoculated with 1% L. lactis ssp. lactis INIA 415-2 and 2% S. thermophilus culture. After ripening for 15 d at 12 degrees C, half of the cheeses from each vat were treated at 400 MPa for 5 min at 10 degrees C. Ripening of high-pressure-treated and untreated cheeses continued at 12 degrees C until d 50. High-pressure treatment of cheese made from milk without the bacteriocin producer accelerated casein degradation and increased the free AA content, but it did not significantly influence the taste quality or taste intensity of the cheese. Addition of the bacteriocin producer to milk lowered the ratio of hydrophobic peptides to hydrophilic peptides, increased the free AA content, and enhanced the taste intensity. The combination of milk inoculation with the bacteriocin producer and high-pressure treatment of the cheese resulted in higher levels of both hydrophobic and hydrophilic peptides but had no significant effect on the free AA content, taste quality, or taste intensity.

Amino Acids↗
PubMed2007

Effects of high pressure treatment on volatile profile during ripening of ewe milk cheese.

The effect of high-pressure treatment on the volatile profile of ewe milk cheeses was investigated. Cheeses were submitted to 200, 300, 400 and 500 MPa at 2 stages of ripening (after 1 and 15 d of manufacturing) and volatile compounds were assayed at 15 and 60 d of ripening. High-pressure treatment altered the balance of volatile profile of cheeses, limiting the formation of acids, alcohols, ketones, aldehydes, and sulfur compounds and enhancing the formation of 2,3-butanedione. In general, cheeses pressurized at 15 d of ripening were more similar to untreated cheeses than those treated at 1 d. Cheeses treated at 300 MPa after 1 d of manufacturing were characterized by higher levels of free amino acids, ethanol, ethyl esters, and branched-chain aldehydes, whereas cheeses treated at 500 MPa after 1 d of manufacturing had lower microbial populations, showed the highest abundance of 2,3-butanedione, pyruvaldehyde, and methyl ketones, and the lowest abundance of alcohols.

Animals↗