Influence of ripening on proteolysis and lipolysis of Murcia al Vino cheese

The aim of this study was to evaluate the influence of five different manufacturers and two ripening periods on the proteolysis and lipolysis patterns of Murcia al Vino goat cheese. The manufacturers significantly affected the water activity (a_w), pH, dry matter and fat content, several nitrogen fractions: water soluble nitrogen (WSN), trichloroacetic acid (12% w/v) soluble nitrogen (TCASN) and phosphotungstic acid (5% w/v) soluble nitrogen (PTASN); also the free amino acid (FAA) and free fatty acid (FFA) contents, with the exception of C_4:0, C_16:0 and C_18:0. Different ripening periods significantly affected the dry matter content, WSN and PTASN and all FAA, except serine.     

Application of linear discriminant analysis to different proteolysis parameters for assessing the ripening of Manchego cheese

Different proteolysis parameters, the nitrogenous fractions and the breakdown of caseins, were determined for Manchego cheeses at different stages of ripening. Linear discriminant analysis was applied to these parameters to ascertain the degree of ripening. Two discriminant functions enabling 100% correct classification of the cheeses into fresh, medium ripe and aged were found.     

Effect of high pressure homogenisation of milk on cheese yield and microbiology, lipolysis and proteolysis during ripening of Caciotta cheese

The principal aim of this work was to compare Caciotta cheeses obtained from cow milk previously subjected to high pressure homogenisation (HPH) at 100 MPa with those produced from raw (R) or heat-treated (P) cow milk. HPH had both direct and indirect effects on cheese characteristics and their evolution during ripening. In particular, HPH treatment of milk induced a significant increase of the cheese yield; moreover, it affected the microbial ecology of both curd and cheese. Compared with the thermal treatment, the HPH treatment resulted in a decrease of about one log cfu/g of yeast and lactobacilli cell loads of the curd. The initial milk treatment also affected the evolution over time and the levels attained at the end of ripening of all the microbial groups studied. In fact, lactobacilli, microstaphylococci and yeast cell loads remained at lower levels in the cheeses obtained from HPH milk with respect to the other cheese types over the whole ripening period. Moreover, HPH of milk induced marked and extensive lipolysis. Cheeses from HPH milk showed the presence of high amounts of free fatty acids immediately after brining. The electrophoretic patterns of the different cheese types showed that Caciotta made from HPH-treated milk was characterized by a more extensive and faster proteolysis as well as a significant modification of its volatile molecule profile. The results obtained and the sensory analysis indicated that HPH treatment of milk was able to differentiate Caciotta cheese or to modify its ripening patterns.     

Caprine cheese with probiotic strains: the effects of ripening temperature and relative humidity on proteolysis and lipolysis

 The effects of ripening temperature, relative humidity and time on chemical and textural characteristics of a 'probiotic' goat's milk cheese were examined. The experimental layout followed a 2³ factorial design, with all possible combinations of 5  °C and 10  °C (ripening temperature), 85% and 95% (ripening relative humidity) and 1 day and 70 days (ripening time). All proteolytic indices measured (water-soluble nitrogen, trichloroacetic acid-soluble nitrogen and phosphotungstic acid-soluble nitrogen) were enhanced with increased ripening temperature to a greater extent than with increased ripening relative humidity; the increase in phosphotungstic acid-soluble nitrogen was the most significant. Free fatty acid concentrations in cheeses were not influenced by ripening relative humidity but increased with ripening temperature and time. A higher ripening temperature and a lower relative humidity gave rise to firmer cheeses. Postulated empirical models have provided a good fit to the experimental data set generated; such models were able to predict a decrease of 25 days in ripening time with no impairment of either proteolytic or lipolytic indices if a cheese were to be ripened at 10  °C (rather than 5  °C) and 95% relative humidity. Received: 23 March 1998     

The effect of fat content on the microbiology and proteolysis in cheddar cheese during ripening dairy foods

We investigated the effect of incremental reduction in fat content, in the range 33 to 6% (wt/wt), on changes in the microbiology and proteolysis of Cheddar cheese, over a 225-d ripening period at 7 degrees C. A reduction of fat content resulted in significant increases in contents of moisture and protein and a decrease in the concentration of moisture in nonfat substance. Reduced fat had little effect on the age-related changes in the population of starter cells. The populations of nonstarter lactic acid bacteria decreased with fat content, and counts in the low fat cheese (6% wt/wt) were significantly lower than those in the full fat cheese (33% wt/wt) at ripening times >1 and <180 d. Proteolysis as measured by the percentage of total N soluble at pH 4.6 or in 70% ethanol decreased significantly as the fat content decreased. However, the content of pH 4.6 soluble N per 100 g of cheese was not significantly influenced by fat content. At ripening times >60 d, the content of 70% ethanol soluble N per 100 g of full fat (33% wt/wt) cheese was significantly lower than that in either the half fat (17% wt/wt) or low fat (6% wt/wt) cheeses. The concentration of AA N, as a percentage of total N, was not significantly affected by fat content. However, when expressed as a percentage of total cheese, amino acid N increased significantly with decreasing fat content. Analysis of pH 4.6 soluble N extracts by reverse phase- and gel permeation HPLC revealed that fat content affected the pattern of proteolysis, as reflected by the differences in peptide profiles.     

Modeling of proteolysis and lipolysis in Iranian white brine cheese

The simultaneous effects of processing variables such as ripening time (20–60 days), ripening temperature (6–10 °C), level of rennet added (1–2 g/100 kg milk) and brine concentration (8–14%, w/v) on the proteolysis, lipolysis and sensory score of Iranian white brined cheese (Feta type) were explored by the means of response surface methodology. The most important effect in proteolytical terms was produced by ripening temperature and ripening time in linear form, but level of rennet added and brine concentration were also significant at the 5% level. In terms of lipolysis, ripening time was dominant factor in both linear and quadratic forms; quadratic effect of ripening temperature was greater than its linear effect.     

Ultrafiltration of cheese milk: Effect on plasmin activity and proteolysis during cheese ripening

Havarti 45+ cheese was manufactured from milk concentrated 1.8–4.6-fold by ultrafiltration (UF) and from normal milk, and the effect of concentration factor on plasmin activity and subsequent proteolysis in cheese during ripening was examined. There was decreased plasmin activity and a reduced rate of proteolysis of α_S2-casein and β-casein in the UF-cheeses, compared with the corresponding controls, independent of concentration factor. The decreased plasmin activity and slower breakdown of α_S2-casein and β-casein in UF-cheeses compared with traditional cheeses can be partly explained by the inclusion of an increased amount of plasmin inhibitors into the UF-cheeses. However, it is suggested that the differences in plasmin activity and proteolysis arise mainly as a result of inactivation of the plasminogen activation system during UF-concentration, due to a combination of time, temperature and the presence of air in the UF-equipment. The effect of milk treatment on the plasminogen activation system should be further investigated.     

成熟温度对Mozzarella干酪蛋白水解和质构的影响

研究了温度为4,7,10℃时对干酪成熟过程中蛋白水解和质地的影响。结果表明,随着干酪成熟温度的升高,成熟期间干酪中可溶性氮与总氮的比值增加较快,干酪的硬度下降速度也较快。说明在较高的成熟温度下,干酪在较短的时间内能够达到成熟的状态。     

The effect of Allium sp. on the extension of lipolysis and proteolysis in Van herby cheese during maturation

The aim of this study was to determine the effect of herb (Allium sp.) on biochemical changes of herby cheese produced in Turkey. Raw cows' milk was used for cheese manufacture. Five groups of cheeses, containing 0 (as control), 0.5, 1, 2, and 3% herb, were produced and coded as K, A, B, C, and D respectively. All cheese groups were ripened at 8 degrees C for 90 days. Samples were taken from cheeses after 3, 15, 30, 60 and 90 days, and analysed for lipolysis (as acid degree value) and proteolysis (water-soluble nitrogen, TCA-soluble nitrogen, and PTA-soluble nitrogen). It was found that lipolysis in herby cheeses increased with increasing herb addition, and the increase in lipolysis degree was significant (P < 0.05) in cheese D. Water-soluble N, TCA-soluble N, and PTA-soluble N as indicator of proteolysis degrees were affected significantly (P < 0.05) by increasing herb ratios.     

The influence of ripening temperature and sampling site on the lipolysis in Reggianito Argentino cheese

The effect of ripening temperature elevation and sampling site on lipolysis in Reggianito Argentino cheese was evaluated. Cheeses ripened at 12 or 18 °C and 85% relative humidity were assayed at 2, 4 and 6 months at two sampling zones (central and external). Samples were analysed to determine physicochemical parameters and the concentration of nine free fatty acids (FFAs) (C_6:0–C_18:2). Myristic, palmitic, stearic and oleic acids were found in higher concentration. While ripening time and temperature significantly affected the concentrations of the nine FFAs analysed, sampling zone significantly affected only two FFAs. Ripening temperature increased the lipolytic process, but it seems to have no effect on the pathways of lipolysis in Reggianito Argentino cheese.     

Seasonal variations in the free fatty acid composition of Manchego cheese and changes during ripening

 This paper examines the evolution of the free fatty acid concentration in Manchego cheese during ripening (up to 150 days) and in two different cheesemaking seasons (autumn and winter). Factorial analysis of the principal components showed that short-chain fatty acids were the variables that best correlated (81% of the total variance explained) with the samples when distributed according to ripening time. These fatty acids likewise differentiated autumn and winter samples. Received: 30 April 1999 / Revised version: 5 July 1999     

Chemometric analysis of proteolysis during ripening of Ragusano cheese

Chemometric modeling of peptide and free amino acid data was used to study proteolysis in Protected Denomination of Origin Ragusano cheese. Twelve cheeses ripened 3 to 7 mo were selected from local farmers and were analyzed in 4 layers: rind, external, middle, and internal. Proteolysis was significantly affected by cheese layer and age. Significant increases in nitrogen soluble in pH 4.6 acetate buffer and 12% trichloroacetic acid were found from rind to core and throughout ripening. Patterns of proteolysis by urea-PAGE showed that rind-to-core and age-related gradients of moisture and salt contents influenced coagulant and plasmin activities, as reflected in varying rates of hydrolysis of the caseins. Analysis of significant intercorrelations among chemical parameters revealed that moisture, more than salt content, had the largest single influence on rates of proteolysis. Lower levels of 70% ethanol-insoluble peptides coupled to higher levels of 70% ethanol-soluble peptides were found by reversed phase-HPLC in the innermost cheese layers and as the cheeses aged. Non-significant increases of individual free amino acids were found with cheese age and layer. Total free amino acids ranged from 14.3 mg/g (6.2% of total protein) at 3 mo to 22.0 mg/g (8.4% of total protein) after 7 mo. Glutamic acid had the largest concentration in all samples at each time and, jointly with lysine and leucine, accounted for 48% of total free amino acids. Principal components analysis and hierarchical cluster analysis of the data from reversed phase-HPLC chromatograms and free amino acids analysis showed that the peptide profiles were more useful in differentiating Ragusano cheese by age and farm origin than the amino acid data. Combining free amino acid and peptide data resulted in the best partial least squares regression model (R(2) = 0.976; Q(2) = 0.952) predicting cheese age, even though the peptide data alone led to a similarly precise prediction (R(2) = 0.961; Q(2) = 0.923). The most important predictors of age were soluble and insoluble peptides with medium hydrophobicity. The combined peptide data set also resulted in a 100% correct classification by partial least squares discriminant analysis of cheeses according to age and farm origin. Hydrophobic peptides were again discriminatory for distinguishing among sample classes in both cases.     

The influence of ripening temperature and sampling site on the proteolysis in Reggianito Argentino cheese

The effects of elevated ripening temperature and sampling site on proteolysis in Reggianito Argentino cheese were evaluated. Cheeses ripened at 12 or 18 °C and 85% relative humidity were analysed at 2, 4 and 6 months in 2 sampling zones (central and external). Samples were analysed to determine the physicochemical and proteolysis parameters through the urea-PAGE of the urea-soluble fraction, RP-HPLC analysis of the water-soluble fraction at pH 4.6, and the free amino acid analysis. Proteolysis was significantly affected by ripening temperature and sampling site. Urea-PAGE analysis showed that elevated temperature increased the degradation of α_s1- and β-casein. The degradation of α_s1-casein was larger in the central zone than in the external one, while β-casein degradation was similar in both zones. The majority peaks detected by RP-HPLC of the water-soluble fraction at pH 4.6 and free amino acids were significantly affected by ripening temperature and sampling site. Glu, His, Val, Leu, and Lys had the higher concentrations. Principal component analysis showed useful groupings when results from chromatograms were studied. In conclusion, the results obtained not only are useful to characterise the ripening of an Argentinean hard cheese, but also to evaluate the effect of an increase of ripening temperature on Reggianito Argentino cheese proteolysis.     

Evolution of lipolysis during the ripening of traditional Feta cheese

Lipolysis was studied during ripening of traditional Feta cheese produced in two small dairies, A and B. The cheeses were made from a thermized mixture of ewes’/goats’ milk by using yoghurt as starter and artisanal rennet from lambs’ and kids’ abomasa (cheese A) or mixed artisanal rennet with calf rennet (cheese B).The acid degree value and the free fatty acids (FFA) contents in both cheeses increased sharply up to 18 d (pre-ripening period at 15 °C) and continued to increase throughout ripening. In both mature cheeses, acetic acid was found at high levels (13–18% of the total FFAs). However, except for this, all FFA contents differed significantly (P < 0.05) between the two cheeses throughout ripening. The levels of individual and total C2:0–C8:0, C10:0–C14:0 and C16:0–C18:2 fatty acids were significantly higher (P < 0.05) in cheese A than in cheese B. Presumably the difference, especially in the C2:0–C8:0 content, was due mainly to the type of the rennet used. Butyric acid was the dominant FFA in cheese A (20% of the total FFAs at 120 d), while the most abundant FFAs in cheese B were capric (18%) and lauric acid (18%). In general, the lipolysis degree of the two cheeses was higher than those reported for the industrially-made Feta cheese.In organoleptic evaluation, cheese A had a piquant taste that was attributed to its high content of butyric acid and showed a significantly (P < 0.05) higher total score than cheese B.     

Accelerated ripening of Cheddar cheese at elevated temperatures

Blocks (20 kg) of Cheddar cheese from a single vat were obtained from a local factory. Half the cheeses were cooled rapidly (15 h) to ripening temperature (8, 12 or 16 °C) and half were cooled slowly over 8 days to the same ripening temperatures. Cheeses were ripened for 9 months at 7 different time/temperature combinations. Ripening temperature had little influence on the number of non-starter lactic acid bacteria in the cheeses after 9 months, although rapid cooling to and ripening at 8 °C drastically reduced the growth rate of these adventitious bacteria. Proteolysis (as determined by urea-polyacrylamide gel electrophoresis; increases in water-soluble N; increases in phosphotungstic acid-soluble N; Cd ninhydrin-reactive amino groups; and reverse-phase HPLC) and lipolysis were accelerated by increasing the ripening temperature and by slow cooling of the cheeses. The rate of ripening was increased or decreased by changing the temperature. Cheeses ripened at 16 °C generally received the highest flavour scores, particularly early during ripening. However, the texture of these cheeses deteriorated after prolonged ripening at 16 °C. Maturation at 12 °C was considered to be optimal for the commercial acceleration of Cheddar cheese ripening.     

凝乳酶对牦牛乳硬质干酪成熟期间蛋白降解的影响

以新鲜牦牛乳为原料,采用小牛皱胃酶、木瓜蛋白酶和微生物凝乳酶制作硬质干酪,探讨凝乳酶种类对牦牛乳硬质干酪成熟期间蛋白质降解的影响。结果表明:三种凝乳酶牦牛乳硬质干酪成熟过程中,不同凝乳酶牦牛乳硬质干酪在成熟期间蛋白质降解能力存在较大差异,总氮（TN）、p H4.6水溶性氮（p H4.6-SN/TN）、12%的三氯乙酸氮（12%TCA-N/TN）、5%磷钨酸氮（5%PTA-N/TN）含量、游离氨基酸均随成熟时间延长不同程度的增加,蛋白氮和酪蛋白氮逐渐降低,多肽氮呈先升高后下降趋势,且微生物凝乳酶降解牦牛乳硬质干酪蛋白能力显著（p<0.05）高于木瓜蛋白酶和小牛皱胃酶。      

Secondary proteolysis in Serra cheese during ripening and throughout the cheese-making season

 Experimental Serra cheeses were manufactured from raw ewe’s milk and thistle flowers following a two-way factorial design. The content of nitrogen soluble in water (WSN), in 2% trichloroacetic acid (2% TCA-N), in 12% TCA (TCA-N) and in 5% phosphotungstic acid (5% PTA-N), and the pH and salt-in-moisture concentration were measured throughout the ripening period (sampling at 0, 7, 21 and 35 days) and the cheese-making season (sampling in November, February and May). Proteolysis in 35-day-old Serra cheese was quantitatively high [average values of 34.6% and 11.9% for WSN/TN (total nitrogen) and 2% TCA-N/TN, respectively], but qualitatively low (average values of 5.8% and 1.2% for 12% TCA-N/TN and 5% PTA-N/TN, respectively). The ratios WSN/TN and 2% TCA-N/TN were lowest for cheeses ripened in February, whereas the ratio 12% TCA-N/TN was highest for cheeses ripened in November. By 35 days of ripening, the average pH and salt-in-moisture concentration values were 5.2 and 4.8%, respectively. No correlation was found to occur between the measured pH or salt-in-moisture concentration and the values of soluble nitrogen fractions throughout the cheese-making season. Received: 14 February 1996/Revised version: 4 June 1996