Effect of high-pressure-processing on the microbiology,proteolysis, texture and flavour of Brie cheese during ripening and refrigerated storage

Brie cheeses were high pressure (HP)-treated at 400 or 600 MPa on days 14 or 21 after manufacture to prevent over-ripening. Lactic acid bacteria and Penicillium camemberti numbers declined markedly after HP treatment. In control cheese pH increased 2.0 units from day 21 to day 60, but less than 0.3 units in HP-treated cheeses. Cheeses treated at 600 MPa showed the maximum concentrations of residual caseins during refrigerated storage and control cheese the minimum concentrations. A 7.6-fold increase in hydrophobic peptides was recorded from day 21 to day 60 in control cheese and 0.8–1.6-fold increases in HP-treated cheeses. The maximum aminopeptidase activity was detected in control cheese, the highest free amino acid concentrations in cheeses treated at 400 MPa. The firmest texture was recorded for cheeses treated on day 14 at 400 or 600 MPa. HP-treated cheeses showed higher flavour quality scores than control cheese from day 60 onwards.     

Using High-Pressure Processing for Reduction of Proteolysis and Prevention of Over-ripening of Raw Milk Cheese

High-pressure-processing (HPP) at 400 or 600 MPa was applied to cheeses made from ewe raw milk, on days 21 or 35 after manufacturing, to reduce proteolysis and prevent over-ripening. The characteristics of HPP and non-pressurized (control) cheeses were compared during ripening at 8 °C until day 60 and further storage at 4 °C until day 240. HPP and control cheeses showed similar pH values throughout ripening, but on day 240 pH values remained 0.4–0.6 units lower for HPP cheeses than for the control cheeses. Casein degradation was significantly retarded in the 600 MPa cheeses. Their α-casein concentration was 48–52 % higher on day 60 and 30–33 % higher on day 240 than in the control cheeses while β-casein concentration was 25–26 % higher on day 60 and 100–103 % higher on day 240. No significant differences in para-κ-casein concentration between cheeses were found on day 60, but on day 240, it was 22–35 % higher in the 600 MPa cheeses than in the control cheese. Hydrophilic peptides, hydrophobic peptides and total free amino acids evolved similarly in HPP and control cheeses during the 60-day ripening period. However, on day 240 hydrophilic peptides were at 34–39 % lower levels in the 600 MPa cheeses than in the control cheeses, hydrophobic peptides at 7–16 % lower levels and total free amino acids at 25–29 % lower levels. Flavour intensity scores increased at a slower rate in HPP cheeses than in the control cheese. Flavour quality declined markedly in the control cheeses during refrigerated storage while it did not vary significantly in 600 MPa cheeses.     

Effect of a bacteriocin-producing Lactococcus lactis strain and high-pressure treatment on the esterase activity and free fatty acids in Hispánico cheese

The effect of milk inoculation with a bacteriocin-producing (BP) culture and of high-pressure (HP) treatment of 15-day-old Hispánico cheeses (400 MPa, 5 min, 10 °C), separately or combined, on the release of intracellular esterases and cheese lipolysis was investigated. Esterase activity and free fatty acids (FFAs) content increased during ripening of Hispánico cheese and palmitic, oleic and stearic acids being the most abundant FFAs. On day 15, the highest esterase activity was recorded for HP-treated BP cheese. The activity for HP-untreated BP cheese was the next highest. No difference in the activities was found between HP-treated and untreated cheeses made without BP culture. Total FFAs on day 15 were at a lower concentration in BP cheeses than in cheeses made without BP culture, probably due to the lower pH values of the former. The rate of total FFA accumulation from day 15 to day 50 was higher in BP cheeses (31.1–32.1% increase) than in cheeses made without BP culture (19.3–21.7% increase). The highest total FFA concentration on day 50 (612 mg kg⁻¹) was found for HP-untreated cheese made without BP culture.     

Effects of high-pressure treatment on free fatty acids release during ripening of ewes' milk cheese

The free fatty acid (FFA) profile of high pressure treated ewes' milk cheeses were studied to assess the effect of pressure treatment on cheese lipolysis. Cheeses were treated at 200, 300, 400 or 500 MPa (2P to 5P) at two stages of ripening (after 1 and 15 days of manufacturing; P1 and P15) and FFA were assayed at 1, 15 and 60 d ripening. On the first day of ripening, 3P1-cheeses showed levels of FFA twice that of the control cheeses. However, no significant differences were found between 3P1 and control cheeses at 60 d ripening. On the contrary, 4P1 and 5P1-cheeses had the lowest total FFA levels. The point at which pressure treatment was applied influenced the FFA profile of cheeses; cheeses pressurized at pressures<400 MPa on the first day of ripening were more similar to untreated cheeses than their homologues treated at 15 d.     

Manufacture of Cheddar cheese from high-pressure-treated whole milk

The cheese-making characteristics of high-pressure (HP)-treated milk were examined. The rennet coagulation time of pasteurised milk decreased after HP treatment at 400 MPa but increased after treatment at 600 MPa. The L-value (whiteness) of milk decreased directly after HP treatment but, over the course of coagulation, whiteness of HP-treated milk increased to the same level as in the control. Cheddar cheese was then manufactured from raw whole milk or whole milk treated by high-pressure (HP) at 400 MPa (HP400) or 600 MPa (HP600) for 10 min at 20 °C. HP treatment of raw milk at 600 MPa resulted in a 3.66 log reduction in the initial counts of non-starter lactic acid bacteria (NSLAB), decreased protein and fat content, as well as a lower pH compared to the control. Furthermore, higher treatment pressures resulted in increased incorporation of β-lactoglobulin into the cheese curd, with parallel increases in yield by 1.23% and 7.78% for HP400 and HP600 cheeses, respectively. Overall, this study showed that the effects of HP treatment on milk proteins increased rennet coagulation times and changes in cheese composition at day 1.Industrial relevanceHigh-pressure treatment is a novel technology which has been applied to a number of commercial food products. In this study, HP-induced changes in milk proteins resulted in increased cheese yields and increased cheese whiteness. In addition, HP treatment significantly reduced the microflora of raw milk cheese. Those attributes could be of interest for both industry and consumer.     

Application of high-pressure treatment on ovine brined cheese: Effect on composition and microflora throughout ripening

Ovine brined cheese was high-pressure (HP) treated at 200 or 500 MPa for 15 min at 20 °C on the 15th day of ripening. Compared to control cheese, HP treatment did not affect significantly (P > 0.05) the pH values, moisture, fat in dry matter, protein in dry matter and salt in moisture contents of cheeses at 90 days. The counts of total aerobic mesophilic bacteria, thermophilic lactococci, thermophilic lactobacilli and non starter lactic acid bacteria (NSLAB) were not affected by HP treatment of cheese at 200 MPa throughout ripening. After 90 days of ripening, the same microbial groups in cheese treated at 500 MPa were about 1.2, 3.6, 2.1 and 4 log units lower than in control cheese respectively. Coliforms were reduced faster at non detectable levels in HP treated cheeses than in control cheese. Regarding the bacterial enzymatic activities in cheese, aminopeptidase activity (Apep) was marginally favoured by both HP treatments. However, its activity was decreased at 90 days due probably to loss in brine. In contrast, lactate dehydrogenase (LDH) activity, following the bacteria cell lysis, was negatively affected by HP treatment at 500 MPa throughout ripening.Industrial relevanceThe data obtained from this work suggest that application of HP treatment under optimized conditions on ovine cheese in brine can be used to reduce effectively the undesirable microbial load in it and to cause moderate enhancement of aminopeptidase activity, without modifying its composition.     

Proteolysis and biogenic amine buildup in high-pressure treated ovine milk blue-veined cheese

Penicillium roqueforti plays an important role in the ripening of blue-veined cheeses, mostly due to lactic acid consumption and to its extracellular enzymes. The strong activity of P. roqueforti proteinases may bring about cheese over-ripening. Also, free amino acids at high concentrations serve as substrates for biogenic amine formation. Both facts result in shorter product shelf-life. To prevent over-ripening and buildup of biogenic amines, blue-veined cheeses made from pasteurized ovine milk were high-pressure treated at 400 or 600 MPa after 3, 6, or 9 wk of ripening. Primary and secondary proteolysis, biogenic amines, and sensory characteristics of pressurized and control cheeses were monitored for a 90-d ripening period, followed by a 270-d refrigerated storage period. On d 90, treatments at 400 MPa had lowered counts of lactic acid bacteria and P. roqueforti by less than 2 log units, whereas treatments at 600 MPa had reduced lactic acid bacteria counts by more than 4 log units and P. roqueforti counts by more than 6 log units. No residual α-casein (CN) or κ-CN were detected in control cheese on d 90. Concentrations of β-CN, para-κ-CN, and γ-CN were generally higher in 600 MPa cheeses than in the rest. From d 90 onwards, hydrophilic peptides were at similar levels in pressurized and control cheeses, but hydrophobic peptides and the hydrophobic-to-hydrophilic peptide ratio were at higher levels in pressurized cheeses than in control cheese. Aminopeptidase activity, overall proteolysis, and free amino acid contents were generally higher in control cheese than in pressurized cheeses, particularly if treated at 600 MPa. Tyramine concentration was lower in pressurized cheeses, but tryptamine, phenylethylamine, and putrescine contents were higher in some of the pressurized cheeses than in control cheese. Differences in sensory characteristics between pressurized and control cheeses were generally negligible, with the only exception of treatment at high pressure level (600 MPa) at an early ripening stage (3 wk), which affected biochemical changes and sensory characteristics.     

Volatile compounds, odor, and aroma of La Serena cheese high-pressure treated at two different stages of ripening

La Serena cheeses made from raw Merino ewe's milk were high-pressure (HP) treated at 300 or 400 MPa for 10 min on d 2 or 50 after manufacture. Ripening of HP-treated and control cheeses proceeded until d 60 at 8°C. Volatile compounds were determined throughout ripening, and analysis of related sensory characteristics was carried out on ripe cheeses. High-pressure treatments on d 2 enhanced the formation of branched-chain aldehydes and of 2-alcohols except 2-butanol, but retarded that of n-aldehydes, 2-methyl ketones, dihydroxy-ketones, n-alcohols, unsaturated alcohols, ethyl esters, propyl esters, and branched-chain esters. Differences between HP-treated and control cheeses in the levels of some volatile compounds tended to disappear during ripening. The odor of ripe cheeses was scarcely affected by HP treatments on d 2, but aroma quality and intensity scores were lowered in comparison with control cheese of the same age. On the other hand, HP treatments on d 50 did not influence either the volatile compound profile or the sensory characteristics of 60-d-old cheese.     

Yield, composition and rheological characteristics of cheddar cheese made with high pressure processed milk

High Pressure (HP) treatment of milk prior to cheese-making was shown to increase the yield of cheese due to increased protein and moisture retention in cheese. Cheeses were made with raw milk or milk treated with high temperature short-time (HTST) pasteurization, and HP treatments at two levels (483 and 676 MPa) at 10 °C, 483 MPa HP at 30 °C, and 483 MPa HP at 40 °C. Cheese yield, total solids, protein, fat and salt contents were evaluated, and fat and protein recovery indices were calculated. Cheeses from HP treatments of 676 MPa at 10 °C and 483 MPa at 30 °C exhibited wet yields of 11.40% and 11.54%, respectively. Protein recovery was 79.9% for HP treatment of 676 MPa at 10 °C. The use of slightly higher pressurization temperatures increased moisture retention in cheese. Visco-elasticity of cheeses was determined by dynamic oscillatory testing and a creep-recovery test. Rheological parameters such as loss (G″) and storage (G′) moduli were dependent on oscillation frequency. At high (173 rad/s) and low (2.75 rad/s) angular frequencies, cheeses made from milk treated at 483 MPa at 10 °C behaved more solid-like than other treatments. Creep tests indicated that cheeses from milk treated with 483 MPa HP at 10 °C showed the smallest instantaneous compliance (J_o), confirming the more solid-like behavior of cheese from the 483 MPa at 10 °C treatment compared to the behavior of cheeses from other treatments. Cheeses made with pasteurized milk were more deformable, exhibited less solid-like behavior than cheeses made with HP treated milk, as shown by the J_o value. With more research into bacteriological implications, HP treatment of raw milk can augment Cheddar cheese yield with better curd formation properties.     

Effect of high pressure on fresh cheese shelf-life

The effect of high pressure (HP; 300 and 400 MPa for 5 min at 6 °C) on physico-chemical, microbial, color, texture and sensorial characteristics of starter-free fresh cheeses stored at 4 and 8 °C was studied. Physico-chemical parameters considered were total solids, fat, total protein, pH, whey loss and water activity. The microbiological quality was studied, on cheeses stored at 4 and 8 °C, by enumerating aerobic mesophilic bacteria, lactococci, psychrotrophic bacteria, Enterobacteriaceae, Escherichia coli, molds and yeasts. Cheeses treated at 300 and 400 MPa, stored at 4 °C, presented a shelf-life of 14 and 21 days, respectively, compared to untreated control cheese, which presented a shelf life of 7 days. On the other hand, HP treatments modified the texture (more firm) and color (more yellow) compared to control cheeses. These changes were detected by instrumental and sensory analysis.     

Cheesemaking with a Lactococcus lactis strain expressing a mutant oligopeptide binding protein as starter results in a different peptide profile

Lactic starters used for cheese manufacture play an important role in the production of bitter peptides and their degradation to non-bitter products. The oligopeptide transport system (Opp) of lactococci is essential for milk peptide utilization. The periplasmic substrate binding protein serves to capture the substrate with high affinity and to deliver it to a membrane-bound complex that translocates it inside the cell. Prt(+)- and Lac(+)-derivatives of MG1363 DeltaoppA strains expressing a wild-type MG1363 OppA or a mutant OppA with a single point mutation at residue 471 (OppA(D471R)) from a plasmid were constructed. These strains were used as lactic starters in cheese manufacture to improve flavour quality by removing hydrophobic peptides from the cheese matrix, through their preferential transport by OppA(D471R). Cheeses made with these strains were not significantly different from control cheeses after 1 day of ripening with respect to bacterial counts, pH and proteolysis, and only slight differences were recorded after 9 and 20 days of ripening. HPLC chromatograms of the hydrophilic and hydrophobic peptides present in the water-soluble fraction of experimental cheeses showed significant differences in peptide content as well as in peak profiles. These results suggest a different peptide utilization in the strain expressing OppA(D471R) and make it suitable for use as starter to improve cheese quality.     

Impact of nisin producing culture and liposome-encapsulated nisin on ripening of Lactobacillus added-Cheddar cheese

This study aimed to evaluate the effects of incorporating liposome-encapsulated nisin Z, nisin Z producing Lactococcus lactis ssp. lactis biovar. diacetylactis UL719, or Lactobacillus casei-casei L2A adjunct culture into cheese milk on textural, physicochemical and sensory attributes during ripening of Cheddar cheese. For this purpose, cheeses were made using a selected nisin tolerant cheese starter culture. Proteolysis, free fatty acid production, rheological parameters and hydrophilic/hydrophobic peptides evolution were monitored over 6 mo ripening. Sensory quality of cheeses was evaluated after 6 mo. Incorporating the nisin-producing strain into cheese starter culture increased proteolysis and lipolysis but did not significantly affect cheese rheology. Liposome-encapsulated nisin did not appear to affect cheese proteolysis, rheology and sensory characteristics. The nisinogenic strain increased the formation of both hydrophilic and hydrophobic peptides present in the cheese water extract. Sensory assessment indicated that acidic and bitter tastes were enhanced in the nisinogenic strain-containing cheese compared to control cheese. Incorporating Lb. casei and the nisinogenic culture into cheese produced a debittering effect and improved cheese flavor quality. Cheeses with added Lb. casei and liposome-encapsulated nisin Z exhibited the highest flavor intensity and were ranked first for sensory characteristics.     

High-Pressure Processing for the Control of Lipolysis,Volatile Compounds and Off-odours in Raw Milk Cheese

Build-up of flavour compounds throughout ripening of raw milk cheeses may result in strong over-ripening notes during refrigerated storage. In order to control the formation of free fatty acids (FFAs) and volatile compounds, and the appearance of off-odours, raw milk cheeses were high-pressure-processed (HPP) 21 or 35 days after manufacture at 400 or 600 MPa. Ripening proceeded at 8 °C until day 60 and, afterwards, cheeses were held at 4 °C until day 240. The effect of HPP on the formation of FFAs and volatile compounds was dependent on pressure level and cheese age at the time of treatment. On day 60, acetic and propionic acids, branched-chain FFAs and short-chain FFAs showed the lowest (p?<?0.05) concentrations in cheeses treated at 400 or 600 MPa on day 21, while medium- and long-chain FFAs were at similar levels in all cheeses. HPP influenced significantly (p?<?0.05) 84 out of the 94 volatile compounds found in cheese. On day 60, the lowest (p?<?0.05) concentrations of acids, alcohols and esters were recorded for cheeses treated at 400 or 600 MPa on day 21, and the lowest (p?<?0.05) concentrations of ketones for cheeses treated at 400 MPa on days 21 or 35. On day 240, all HPP cheeses showed lower (p?<?0.05) concentrations of aldehydes, esters and, particularly, sulphur compounds than control cheese, which exhibited putrid and rancid off-odours from day 120 onwards. Principal component analysis combining FFAs and volatile compounds discriminated 240-day control cheese from 120-day control cheese and both from the rest of cheeses.     

High-pressure effects on the microstructure, texture, and color of white-brined cheese

Abstract: White‐brined cheeses were subjected to high‐pressure processing (HPP) at 50, 100, 200, and 400 MPa at 22 °C for 5 and 15 min and ripened in brine for 60 d. The effects of pressure treatment on the chemical, textural, microstructural, and color were determined. HPP did not affect moisture, protein, and fat contents of cheeses. Similar microstructures were obtained for unpressurized cheese and pressurized cheeses at 50 and 100 MPa, whereas a denser and continuous structure was obtained for pressurized cheeses at 200 and 400 MPa. These microstructural changes exhibited a good correlation with textural changes. The 200 and 400 MPa treatments resulted in significantly softer, less springy, less gummy, and less chewy cheese. Finally, marked differences were obtained in a* and b* values at higher pressure levels for longer pressure‐holding time and were also supported by ΔE* values. The cheese became more greenish and yellowish with the increase in pressure level. Practical Application: The quality of cheese is the very important to the consumers. This study documented the pressure‐induced changes in selected quality attributes of semisoft and brine‐salted cheese. The results can help the food processors to have knowledge of the process parameters resulting in quality changes and to identify optimal process parameters for preserving pressure‐treated cheeses.     

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°C, half of the cheeses from each vat were treated at 400 MPa for 5 min at 10°C. Ripening of high-pressure-treated and untreated cheeses continued at 12°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.     

Proteolysis, lipolysis, volatile compounds, texture, and flavor of Hispánico cheese made using frozen ewe milk curds pressed for different times

Hispánico cheese is manufactured in Spain from a mixture of cow and ewe milk. Production of ewe milk varies throughout the year, with a peak in spring and a valley in summer and autumn. To overcome this seasonal shortage, curd from spring ewe milk may be frozen and used for cheese manufacture some months later. In the present work, ewe milk curds pressed for 15, 60, or 120 min were held at −24°C for 4 mo, thawed, cut to 1-mm pieces, and mixed with fresh cow milk curd for the manufacture of experimental Hispánico cheeses. Control cheese was made from a mixture of pasteurized cow and ewe milk in the same (80:20) proportion. Cheeses, made in duplicate experiments, were analyzed throughout a 60-d ripening period. No significant differences between cheeses were found for lactic acid bacteria counts, dry matter content, hydrophilic peptides, 47 out of 68 vol.tile compounds, texture, and flavor characteristics. On the other hand, differences of minor practical significance between experimental and control cheeses, unrelated to the use of frozen ewe milk curd or the pressing time of ewe milk curd, were found for pH value, aminopeptidase activity, proteolysis, hydrophobic peptides, free amino acids, free fatty acids, and the remaining 21 vol.tile compounds. It may be concluded that the use of frozen ewe milk curd in the manufacture of Hispánico cheese does not alter its main characteristics.     

Proteolysis during the ripening of goats’ milk cheese made with plant coagulant or calf rennet

Powdered plant coagulant (PPC) obtained from the cardoon (Cynara cardunculus) was compared with calf rennet (CR) for the manufacture of goats’ milk cheese, by determining difference in the proteolysis throughout ripening. There were no substantial differences between the compositions of cheeses made using the two types of coagulants. However, cheeses manufactured with PPC exhibited higher levels of pH 4.6-SN than cheese made using CR. The extent of breakdown of α_s-casein, as measured by urea-PAGE, was greater in cheese made using PPC than cheese made using CR. The formation of hydrophobic peptides and the ratio of hydrophobic/hydrophilic peptides throughout the ripening were higher in cheeses made with PPC than in cheeses made with CR. Principal component analysis (PCA) of peak heights of RP-HPLC peptide profiles of the ethanol-soluble and ethanol-insoluble fractions distributed the samples according to the coagulant used in their manufacture. Quantitative differences in several peptides were evident between the two types of cheese.     

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.     

High Hydrostatic Pressure for Accelerating Ripening of Goat's Milk Cheese: Proteolysis and Texture

High hydrostatic pressurization is proposed for cheese ripening acceleration. Several treatments were used for accelerating ripening of goat's milk cheese: 50 MPa / 72 h, 400 MPa / 5 min and 400 MPa / 5 min followed by 50 MPa / 72 h all at 14 °C. Moisture content and pH were higher in 400 MPa treatments compared to the others. By measuring proteolysis indexes, 400 MPa treatments were found to accelerate ripening (14 d in contrast to 28 d conventionally) due to enhanced enzyme activity from inoculated starter culture. Sensory analysis indicated bitter notes in the accelerated ripened cheese. Pressurized cheeses were less crumbly and more elastic than control.     

Ripening Profiles of Goat Cheese Produced from Milk Treated with High Pressure

Goat cheeses were made from pasteurized (72 °C, 15 s) and high-pressure (HP)-treated milk (500 MPa, 15 min, 20 °C). At 45 days of ripening, cheeses made from both types of milk were similar in moisture, quality, electrophoretic profiles, water-soluble nitrogen, and total free fatty acid contents. Cheeses made from HP-treated milk had higher pH and salt, matured more quickly, as determined by formation of total free amino acids, and developed strong flavors. Reverse-phase high-performance liquid chromatography showed differences between the peptide profiles of the cheeses. Differences in small peptides and free amino acids indicated a higher extent of proteolysis in cheeses made from HP-treated milk.