The influence of milk pasteurization temperature and pH at curd milling on the composition, texture and maturation of reduced fat cheddar cheese

Reduced fat milks were pasteurized, for 15 s, at temperatures ranging from 72 to 88°C to give levels of whey protein denaturation varying from ˜ 3 to 35%. The milks were converted into reduced fat cheddar cheese (16–18% fat) in 500 litre cheese vats; the resultant cheese curds were milled at pH values of 5.75 and 5.35. Raising the milk pasteurization temperature resulted in impaired rennet coagulation properties, longer set-to-cut times during cheese manufacture, higher cheese moisture and moisture in the non-fat cheese substance, lower levels of protein and calcium and lower cheese firmness. Increasing the pH at curd milling from 5.35 to 5.75 affected cheese composition and firmness, during ripening, in a manner similar to that of increasing milk pasteurization temperature. Despite their effects on cheese composition and rheology, pasteurization temperature and pH at curd milling had little influence on proteolysis or on the grading scores awarded by commercial graders during ripening over 303 days .     

地衣芽孢杆菌凝乳酶对切达干酪成熟过程中蛋白水解的影响

利用地衣芽孢杆菌凝乳酶制作切达干酪和切达干酪类似物,分析干酪成熟过程中各蛋白水解指标的变化规律,以揭示地衣芽孢杆菌凝乳酶对切达干酪成熟过程中蛋白水解的影响。结果表明,CDF组（添加地衣芽孢杆菌D3.11凝乳酶所制切达干酪）、CD3组（添加地衣芽孢杆菌D3.11凝乳酶但未添加发酵剂制成的干酪类似物）和CCF组（添加商品凝乳酶所制切达干酪）干酪蛋白含量、pH 4.6-可溶性氮、12%三氯乙酸-可溶性氮、5%磷钨酸-可溶性氮、总游离氨基酸含量均随着成熟时间延长呈显著增加趋势,并且成熟期间CDF组干酪均显著高于CCF组干酪（P＜0.05）;十二烷基硫酸钠-聚丙烯酰氨凝胶电泳分析表明,CDF组干酪α-酪蛋白水解程度较大;pH 4.6-可溶性肽段分析表明,随着干酪的成熟,总肽含量呈先增加后下降趋势,但疏水性肽与亲水性肽的比值呈持续下降趋势,在成熟第6个月时,CDF组、CD3组和CCF组干酪疏水性肽与亲水性肽比值分别为2.668、2.822、3.788。主成分分析表明,3 组干酪的蛋白水解程度与成熟度呈正相关,与疏水性肽和亲水性肽的比值呈负相关。以上结果表明,利用地衣芽孢杆菌凝乳酶制作的干酪蛋白水解度更高,但其疏水性肽比例较小,研究结果可为地衣芽孢杆菌凝乳酶在干酪生产中的应用提供理论依据。     

Effects of changes during ripening and frozen storage on the physicochemical and sensory characteristics of Cabrales cheeses

The physicochemical and organoleptic characteristics of two batches of Cabrales cheese, stored at −20°C for 4 and 8 months, respectively, were studied during subsequent ripening. Frozen storage did not result in significant alterations in overall compositional, rheological and sensory properties or the level of lipolysis. The extent of proteolysis was slightly lower in the cheeses frozen prior to ripening.     

Use of high pressure treatment to attenuate starter bacteria for use as adjuncts for Cheddar cheese manufacture

Attenuated starter bacteria cannot produce acid during cheese manufacture, but contain enzymes that contribute to cheese ripening. The aim of this study was to investigate attenuation of starter bacteria using high pressure treatment, for use in combination with a primary starter for Cheddar cheese manufacture, and to determine the effect of such adjunct cultures on secondary proteolysis during ripening. Lactococcus lactis ssp. cremoris HP and L. lactis ssp. cremoris 303 were attenuated by pressure treatment at 200 MPa for 20 min at 20 °C. Cheddar cheese was manufactured using untreated cultures of both these starter strains, either alone or in combination with their high pressure-treated equivalents. High pressure-treated starters did not produce acid during cheese manufacture and starter counts in cheeses manufactured using high pressure-treated starter did not differ from those of the controls. Higher levels of cell lysis were apparent in cheese manufactured using high pressure-treated strains than in the controls after 26 d of ripening. Small differences were observed in the peptide profiles of cheeses, analysed by reversed-phase HPLC; cheeses manufactured using high pressure-treated starters also had slightly higher levels of amino acids than the relevant controls. Overall, addition of high pressure-treated starter bacteria as a secondary starter culture accelerated secondary proteolysis in Cheddar cheese.

Industrial relevance

Attenuated starters provide extra pool of enzymes, which can influence cheese ripening, without affecting the cheese making schedule. This paper presents an alternative method for attenuation of starter bacteria using high pressure treatment and their subsequent use to accelerate secondary proteolysis in Cheddar cheese during ripening.     

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

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

Effects of ripening conditions on the texture of Gouda cheese

Summary The effects of ripening temperature, type of packaging film and storage period before packaging were related to the degree of proteolysis and the texture of Gouda cheese, so as to determine the optimum ripening conditions. Gouda cheeses from a local plant were subjected to different ripening conditions. A factorial design of 2³× 5 was used, where the three factors selected in two levels were: (1) time of storage before packaging, 4 and 10 days, (2) ripening temperature, 10 and 20 °C and (3) plastic film, BK1 and BK5 (Grace, Quilmes, Argentina). Ripening time was a fourth factor analyzed; sampling times were 15, 25, 35, 49 and 70 days after production. Cheeses traditionally ripened (without packaging) were also analyzed. Water content and pH were determined. Nonprotein nitrogen (soluble in 12% trichloracetic acid (TCA)) was quantified by the Kjeldhal method. Cheese texture was analyzed by compression and relaxation tests which were done by using an Instron Universal Testing Machine (Instron Corp., Canton, MA, USA). The pH and water content of cheeses which ripened at 20 °C were lower than the corresponding ones ripened at 10 °C. Only ripening time and temperature had a significant effect on water content, nonprotein nitrogen concentration and rheological parameters. Results show that texture properties of Gouda cheese ripened in plastic films with low gaseous permeability are similar to those of traditionally ripened Gouda. Texture development was accelerated by increasing the storage temperature.     

Influence of vegetable and animal rennet on proteolysis during ripening in ewes’ milk cheese

The renewed interest in using enzymes from thistles of the genus Cynara in the making of traditional ewes’ milk cheese prompted us to investigate the effect of vegetable and animal rennet on proteolysis during ripening of Los Pedroches cheese. Casein hydrolysis was found to be much more extensive and faster in cheese made by using vegetable rennet (the amount of soluble nitrogen at 60, 80 and 100 days of ripening was more than 28% greater than that in cheese produced using animal rennet). The levels of insoluble Tyr and Trp were higher in cheese produced with vegetable rennet. PAGE, using gels containing 7 M urea, revealed decreased contents in residual α_s-CN and β-CN, as well as markedly increased levels of the more mobile components in cheese produced from vegetable rennet at the end of ripening. On the other hand, the degree of proteolysis in terms of NPN or its main components (peptides, amino acids and ammonia) was similar in cheese produced using animal or vegetable rennet.     

Effect of high-pressure treatments on proteolysis and texture of ewes’ raw milk La Serena cheese

La Serena cheeses, made from Merino ewes’ raw milk, were high-pressure (HP)-treated at 300 or 400 MPa for 10 min at 10 °C, on days 2 or 50 of ripening. Cheeses treated by HP on day 2 showed higher pH values than control cheese on day 3, but cheeses treated by HP on days 2 or 50 and control cheese had similar pH values on day 60. Breakdown of caseins was delayed by HP treatment of cheeses on day 2. Cheeses treated by HP on day 2 showed higher levels of hydrophilic peptides, lower levels of hydrophobic peptides, lower hydrophobic peptides: hydrophilic peptides ratios, and higher total contents of free amino acids than those of control cheese. HP treatment of cheese on day 50 scarcely affected proteolysis of 60-day-old cheeses. Fracturability, hardness and elasticity values of cheeses treated by HP on day 2 were higher than those of control cheese and of cheeses treated on day 50. Cheeses treated at 400 MPa on day 2 received the lowest scores for quality of taste from panellists, whereas the rest of HP-treated cheeses did not differ from control cheese.     

Kinetics of Casein Degradation during Ripening of Fynbo Cheese Salted with NaCl/KCl Brine

First-order kinetics with respect to the α_s1-casein concentration was used to study casein degradation during low-fat Fynbo cheese ripening. Effects of partial NaCl replacement by KCI during cheese salting were studied by statistical treatment of the casein degradation results. Four zones from cheeses at 1, 5, 10, 20, and 30 ripening days were analyzed by a polyacrylamide gel electrophoresis method. Similar kinetic parameters were obtained for a cheese salted with a NaCl/KCl brine and for a control cheese during ripening. Results were more affected by salt concentration than by salt substitution. KCl did not strongly influence kinetics of Fynbo cheese proteolysis.     

The effect of plasmin activity and cold storage of cheese milk on the composition, ripening and functionality of mozzarella-type cheese

The effects of casein hydrolysis by plasmin on composition, ripening and functionality in mozzarella-type cheese were studied. Milk was stored at 4°C for 0, 24 and 48 h, with and without added plasmin, before batch pasteurization and the manufacturing of mozzarella-type cheese. In parallel experiments, the effects of incubation of milk with added plasmin for 2 h at 37°C prior to cheese manufacture and of an increase in the level of rennet added during cheesemaking were studied. Plasmin addition adversely affected the rennet coagulation properties of the milk and led to increased (two- to three-fold) plasmin activity in cheese and greater plasmin-associated hydrolysis of β-casein to γ-caseins during ripening. Melted cheese flow rate and stretchability increased over ripening and melting time decreased, but functional properties were not affected by either plasmin activity or cold storage. Overall, neither plasmin activity (at the levels studied) nor cold storage greatly affected the composition or functionality of mozzarella-type cheese.     

Study of the chemical composition,proteolysis, volatile compounds,and textural properties of industrial and traditional Beaten (Bieno sirenje) ewe milk cheese

The objective of this study was to determine the gross composition, proteolysis, and volatile and texture profiles during ripening of industrial (IND) and traditional (TRD) Beaten (Bieno sirenje) cheeses made by using ewe milk. In the course of the analyses, statistical differences were determined in some physicochemical parameters, nitrogen fractions, and total free amino acid levels between TRD and IND types of cheese. Higher levels of proteolysis were observed in IND cheeses than in TRD cheeses during ripening. Levels of residual β- and α_s-caseins were 72.2 and 48.7%, respectively, in 180-d-old TRD cheeses. However, the residual levels were 52.8% for β-casein and 18% for α_s-casein in IND cheeses. Similar differences were noted for the reversed-phase HPLC peptide profiles of 2 types of cheeses. Also, higher concentrations of peptides were eluted in IND cheeses than in TRD cheeses during ripening. A total of 73 volatile compounds, including alcohols (16), esters (17), acids (14), terpenes (7), ketones (5), aldehydes (4), and miscellaneous (10) were identified. The IND cheeses contained higher levels of carboxylic acids, esters, alcohols, and terpenes than the TRD cheeses; however, the same levels of methyl ketones were determined in the 2 types of cheeses at the end of ripening. These may be due to some differences (e.g., pasteurization and scalding temperature, among other factors) in the manufacture of the 2 types of Beaten cheeses. The textural profile of Beaten cheeses showed that TRD production method resulted in firmer, less fracturable, and stiffer cheeses than the IND production method. In conclusion, the results suggest that the use of industrial production method (pasteurization of cheese milk and curd scalding at 70°C) in the manufacture of Beaten ewe milk cheese enriched the volatile profile of the cheese.     

Effects of Penicillium roqueforti and whey cheese on gross composition,microbiology and proteolysis of mould‐ripened Civil cheese during ripening

Four different types of mould‐ripened Civil cheese were manufactured. A defined (nontoxigenic) strain of a Penicillium roqueforti (SC 509) was used as the secondary starter with and without addition of the whey cheese (Lor); in parallel, secondary starter‐free counterparts were manufactured. Chemical composition, microbiology and proteolysis were studied during the ripening. The incorporation of whey cheese in the manufacture of mould‐ripened Civil cheese altered the gross composition and adversely affected proteolysis in the cheeses. The inoculated P. roqueforti moulds appeared to grow slowly on those cheeses, and little proteolysis was evident in all cheese treatments during the first 90 days of ripening. However, sharp increases in the soluble nitrogen fractions were observed in all cheeses after 90 days. Microbiological analysis showed that the microbial counts in the cheeses were at high levels at the beginning of ripening, while their counts decreased approximately 1–2 log cfu/g towards the end of ripening.     

Evaluation of cheese authenticity and proteolysis by HPLC and urea–polyacrylamide gel electrophoresis

Chromatographic and electrophoretic methods have been established as useful tools in characterising cheese ripening and in the detection of milk adulteration. The purpose of this work was to evaluate casein proteolysis of cheeses made from bovine, ovine or mixtures of bovine and ovine milks, as well as ovine cheese authenticity, for 30 days of ripening by HPLC and urea–polyacrylamide gel electrophoresis.Complementary information was obtained by both techniques when applied to the study of casein proteolysis during 30 days of ripening of ovine milk cheeses, ovine milk cheeses with 10% and 20% of bovine milk and bovine milk cheeses, manufactured according to the traditional Terrincho technology. For ovine cheeses, α-casein was the fraction that showed the higher degradation during cheese ripening. A similar behaviour was observed for ovine milk cheese with 10% of bovine milk. The profile for ovine milk cheese with 20% of bovine milk was more similar to that obtained for bovine cheese. Concerning bovine milk cheeses, electrophoresis was the most sensitive technique for the evaluation of proteolysis in these cheeses.Ten and 20% of bovine milk could be detected in ovine milk cheeses by urea–polyacrylamide gel electrophoresis and HPLC, respectively, even after 30 days of ripening.     

瑞士乳杆菌对契达干酪成熟期间所产ACE抑制肽的影响及其消化稳定性

为明确瑞士乳杆菌对契达干酪中血管紧张素转换酶（angiotensin-converting enzyme,ACE）抑制肽活性的影响,以蛋白质水解度和ACE抑制率为指标,与干酪乳杆菌组、鼠李糖乳杆菌组和空白组干酪进行对照,研究瑞士乳杆菌对干酪成熟期间蛋白质水解及ACE抑制活性的影响,并对ACE抑制活性最高时期的干酪进行消化稳定性研究。结果表明：成熟期间,3 组益生菌干酪的活菌数无明显差异（P＞0.05）,但均高于空白组;益生菌干酪的蛋白质水解程度和ACE抑制活性显著高于空白组（P＜0.05）,其中瑞士乳杆菌干酪的蛋白质水解程度最强,活性最高（79.71%）。模拟消化后,瑞士乳杆菌干酪活菌数降低14.30%,ACE抑制活性显著增加（P＜0.05）,达到86.06%,多肽质量浓度增加至2.81 mg/mL;研究不同分子质量超滤组分消化后的ACE抑制活性发现,其中大于10 kDa的多肽活性升高,小于10 kDa的活性下降。此外,添加瑞士乳杆菌不影响干酪的整体可接受性。因此,瑞士乳杆菌能促进干酪ACE抑制肽的产生并提高其活性,消化后活性的升高主要与大分子肽的降解有关。     

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.     

Proteolysis and microstructure of Piacentinu Ennese cheese made using different farm technologies

The aim of this study was to provide the biochemical and structural characterization of Piacentinu Ennese cheese and to evaluate the impact of different farm technologies on cheese proteolysis and microstructure. Fifteen cheeses were manufactured according to traditional technology, i.e., from raw milk and farmhouse rennet in the absence of starter culture. Pasteurized milk, commercial rennet, and starter were used for production of 20 nontraditional cheeses. Proteolysis in Piacentinu Ennese cheese was monitored during a 2- to 10-mo ripening time. Low rates of overall proteolysis were observed in cheese, as percentages of total N soluble at pH 4.6 and in 12% trichloroacetic acid were about 11.40 and 8.10%, respectively, after 10 mo of age. Patterns of primary proteolysis by urea-PAGE showed that alpha(s)-caseins were degraded to a larger extent than were beta-caseins, although a considerable amount of both caseins was still intact after 10 mo. Reversed phase-HPLC analysis of the cheese peptide fractions showed a slow decrease in the levels of hydrophobic peptides coupled to increasing levels of hydrophilic compounds as the cheese aged. The structural characteristics of Piacentinu Ennese cheese were evaluated by scanning electron microscopy after 2, 4, and 6 mo of age. The micrographs showed a sponge-like structural network with a well-distributed system of empty spaces, originally occupied by whey and fat. The microstructure changed during cheese ripening to become more compact with cavities of smaller size. Farm technology significantly affected cheese proteolysis and microstructure. Nontraditional cheeses had higher levels of pH 4.6-soluble N and showed a larger hydrolysis of alpha(s)-casein fractions by urea-PAGE analysis than did traditional cheeses. Large differences between cheese-types also concerned the patterns of secondary proteolysis. Nontraditional cheeses had higher levels of 12% trichloroacetic acid-soluble N and showed larger proportions of free amino acids and hydrophilic peptides in the HPLC profiles of the corresponding 70% ethanol-soluble N fraction than traditional cheeses. Nontraditional cheeses also had a more open structure with a coarser and less continuous appearance than did traditional cheeses. A large amount of variability in cheese proteolysis and structure within nontraditional treatment reflected farm-dependent changes in manufacturing conditions related to the use of various types of rennet and starter.     

Size-Exclusion HPLC Separation of Bitter and Astringent Fractions from Cheddar Cheese Made with Added Lactobacillus Strains to Accelerate Ripening

Lactobacillus strains were added as an adjunct to the regular lactic starter in Cheddar cheese manufacture in order to accelerate ripening. Microbial cheese proteolysis resulted in the release of free amino acids which were extracted with the astringent and bitter fractions and separated by size-exclusion and reversed-phase HPLC chromatography. Lactobacillus strains generally increased the degree of proteolysis. L. plantarum and L. brevis produced off-flavors possibly due to an accumulation of medium-size peptides. The control cheese (without lactobacilli) had the most peptides with a mean molecular- weight of < 1000 daltons and had a flavor described as slightly bitter. Addition of L. casei-casei L2A accelerated ripening and yielded a well-aged Cheddar cheese without any bitterness even after 7 months at 6°C.     

Ripening of Camembert-type cheese made from caprine milk using calf rennet or kid rennet as coagulant

Camembert-type cheese was made from caprine milk using either calf rennet or kid 'Grandine' rennet as coagulant. The pH of all cheeses increased throughout ripening and levels of pH 4.6-soluble nitrogen increased from 8.1 to 18.2% of total nitrogen (TN) and from 6.9 to 20% TN for the cheeses made using calf rennet and kid rennet, respectively. Degradation of β-casein, measured by urea–polyacrylamide gel electrophoresis, and total and free amino acids were greater in the cheese made using kid rennet. Production of peptides, analysed by high performance liquid chromatography (HPLC), was slightly more extensive in the Camembert-type cheese made using calf rennet as coagulant. In general, a higher degree of proteolysis was found in Camembert-type cheese made from caprine milk using kid rennet than in cheese made using calf rennet as coagulant.     

Ripening of cheese made from full concentrated milk retentate with and without peptidase addition

The aim of this study was to determine ripening of cheese made from full concentrated (FC) milk retentate with and without peptidase addition. No free amino acids (FAAs) were found in FC cheese at the end of ripening. However, added peptidase increased FAA formation. Protein and peptide profile analysis showed that FAA and small peptides increased during ripening and therefore some secondary proteolysis occurred. Added peptidase increased D‐lactic acid formation during ripening of cheeses. This kind of changes in lactose fermentation should be considered during developing the making cheese with different enzyme addition.     

Kinetics of Proteolysis of β-Casein During Ripening of Fynbo Cheese Salted with NaCl or NaCl/KCl and Ripened at Different Temperatures

ABSTRACT: The proteolysis of β-casein during ripening of low-fat Fynbo cheese was studied using 1st-order kinetics to improve the knowledge of the p-casein hydrolysis in Fynbo cheeses salted with NaCl or NaCl/KCl and ripened at different temperatures. Effects of ripening temperature, partial replacement of NaCl by KCl during cheese salting, and total salt concentration were evaluated. Central and external zones from cheeses at 1, 5, 10, 20, 30, 60, and 90 ripening days were analyzed by polyacrylamide gel electrophoresis. No significant differences in the kinetic parameters were observed between cheeses salted with NaCl and those salted with a NaCl/KCl brine. Kinetic constants were significantly affected by region within cheese and ripening temperature. Kinetic constant values were in the range of 0.004/d to 0.018/d, and the activation energy of the reaction was approximately 19 kcal/gmol.