Effects of coagulant type on the physicochemical and organoleptic properties of Kashar cheese

Kashar cheeses were manufactured using different coagulants (calf rennet, chymosin derived by fermentation and proteases from Rhizomucor miehei and Cryphonectria parasitica) and ripened for 90 days. Use of different coagulants did not influence the dry matter, fat, protein, salt, pH, titratable acidity, total free fatty acids and texture profile analyses. The levels of water‐soluble nitrogen, 12% trichloroacetic acid‐soluble nitrogen, and for 5% phosphotungstic acid‐soluble nitrogen, the sensory properties were significantly influenced by the use of different coagulants. β‐casein was more hydrolysed in the cheese manufactured using protease from Cryphonectria parasitica than the other cheeses during 90 d of ripening.     

Casein breakdown during ripening of Idiazabal cheese: influence of starter and rennet type

The objective of this work was to determine the effect of starter and rennet type on casein breakdown during Idiazabal cheese ripening. Four batches of cheeses were manufactured with two rennets, commercial calf rennet and artisanal lamb rennet, and the use of natural flora or a commercial starter. Electrophoretic analysis of cheese samples showed six bands identified as α_s1‐, α_s2 + β‐, α_s1‐I‐, γ₁‐, β‐I‐ and para‐κ‐casein. As expected, the casein breakdown during cheese ripening was considerably affected by rennet type and the use of a commercial starter. The artisanal lamb rennet produced a higher hydrolysis of casein fractions than the commercial calf rennet, probably owing to its high percentage of chymosin (around 78%). The effect of addition of starter on proteolysis was dependent on the casein fractions generated by artisanal lamb rennet or commercial calf rennet. © 2000 Society of Chemical Industry     

基于多元线性回归对Tilsit型干酪成熟期的研究

根据Tilsit干酪加工方法制作生干酪样品,在90 d成熟内测定其总氮、水溶性氮(WSN)、三氯乙酸水溶性氮(TCA-SN)、磷钨酸水溶性氮(PTA-SN)等含量,研究成熟过程中蛋白质水解程度。上述指标,除磷钨酸水溶性氮,其余均与干酪成熟期呈现一定的线性相关性(p<0.05),该多元线性回归模型可以用于鉴定和预测Tilsit型干酪的成熟期。     

Effects of Packaging Materials and Filling Methods on Selected Characteristics of Otlu (Herby) Cheese

In this research, two kinds of filling methods (block and pieced) in two container types (pot and plastic) were used for the preparation of Otlu cheeses. The uses of different containers influenced (P < 0.01) acidity, pH, dry matter, fat contents and water soluble nitrogen. Degradation of α?s₁- and β-casein were higher in pieced cheeses in pot (P1F2). Also, breakdown products of casein were higher in P1F2 cheeses compared to others. The amounts of α?s₁- and β-caseins of all treatments decreased during ripening, while the amounts of α?s₁-I casein and breakdown products increased. The acceptability of cheeses in block form was higher.     

Proteolysis and texture changes of a Spanish soft cheese (‘Torta del Casar’) manufactured with raw ewe milk and vegetable rennet during ripening

Proteolysis and textural changes of the Spanish ewe raw milk soft cheese of the Protected Designation of Origin Torta del Casar were studied in four different stages of ripening, with 1, 30, 60 and 90 days. In general, proteolysis in Torta del Casar cheese was weak at 1 and 30 days and it was more intense between the 30–60 days of ripening. Soluble nitrogen non‐protein nitrogen, polypeptide N and free amino acids values significantly increased during cheese ripening. Protein and casein nitrogen decreased significantly after 60 days of ripening resulting in the increase of the other nitrogen fractions measured. Caseins changes determined by capillary zone electrophoresis showed that proteolysis of β‐casein occurred faster than αs1‐casein but the latter suffered higher proteolytic degradation at the end of ripening (day 90). This pattern of degradation of caseins is reversed in other cheeses made with animal rennet. Texture analysis showed that firmness and consistency decreased along ripening while adhesiveness increased. Highly significant correlations were found between textural parameters, residual caseins levels and nitrogen fractions during maturation, which shows the importance of proteolytic changes for an optimal texture formation.     

Influence of rennet milk-clotting activity on the proteolytic and sensory characteristics of an ovine cheese

Roncal cheese (regulated by an Apellation of Origin) is a traditional hard cheese manufactured from raw ewe's milk in the region of Navarre in Spain. Roncal cheeses, manufactured using two lamb rennets with different milk-clotting activity levels, were evaluated to compare their chemical, proteolytic, and sensory characteristics. A preliminary study of samples of lamb rennets indicated that a large proportion of such rennets did not fulfil current microbiological requirements and likewise revealed considerable variation in the milk-clotting activity of the samples examined. Trends in the overall physicochemical parameter values (pH, dry matter, fat, and protein) were similar in both cheese batches. Proteolysis of the nitrogen fractions was observed to take place at a faster rate in the cheeses made using the rennet with the higher milk-clotting activity (soluble nitrogen, non-protein nitrogen, and amino acid nitrogen values around 13–20% higher than in the cheeses made using the rennet with the lower milk-clotting activity after 180 days of ripening). Urea-PAGE electrophoretic analysis of the caseins from the cheeses manufactured using both types of rennet showed that the β-caseins were less susceptible to proteolysis than the α_s-caseins. The effect of the different milk-clotting activity levels was most pronounced on the α_s-caseins, in which the rennet with the higher milk-clotting activity gave higher breakdown. Nevertheless, the differences in the proteolysis rates did not yield any appreciable sensory differences.     

Proteolysis in Manchego-type cheese salted by brine vacuum impregnation

A new salting procedure based on the brine vacuum impregnation of porous products was tested on Manchego-type cheese and compared with conventional brine immersion. Its effect on cheese proteolysis throughout a 90-d ripening period was determined. Three cheese regions were evaluated (the rind, the middle, and the internal regions). The parameters analyzed were total N, water-soluble N, soluble N in trichloroacetic acid and soluble N in phosphotungstic acid by using the Kjeldahl method, casein profile by urea-PAGE, and peptide profile of the water soluble nitrogen extract by reverse-phase HPLC. Free amino acid formation was monitored with a spectrophotometric method by using a Cd-ninhydrin reagent. Globally, proteolysis was significantly affected by ripening stage (increasing throughout all the maturation period studied) and cheese region (rind showed a proteolysis pattern different from the middle and internal regions). The salting procedure only affected cheese proteolysis in the rind, whereas conventional brine-salted cheeses showed lower proteolysis than vacuum-impregnated cheeses.     

Multiparametric analysis of cheese using single spectrum of laser-induced breakdown spectroscopy

The potential of using laser-induced breakdown spectroscopy (LIBS) for prediction of chemical quality/control parameters in a cheese matrix was investigated. Traditional methods are usually time-consuming and require complex pre-treatments, the use of toxic chemicals and laboratories and personnel equipped with these analytical skills. In this study, 11 different chemical analyses commonly performed on cheese samples were conducted using traditional methods on 82 full-fat, white pickled and ripened cheese samples. Additionally, these parameters of interest were correlated with multi-elemental spectra of LIBS using partial least squares regression. The results obtained for moisture, dry matter, salt, total ash, total protein, pH, fat, acidity, water soluble nitrogen, trichloroacetic acid soluble nitrogen, and phosphotungstic acid soluble nitrogen (R_cal² = 0.964, 0.959, 0.970, 0.973, 0.952, 0.971, 0.733, 0.762, 0.714, 0.633, and 0.707, respectively) indicated that LIBS could be used as an alternative or complementary method in quality control of cheese.     

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.     

The hydration behaviour of rennet caseins in calcium chelating salt solution as determined using a rheological approach

Rennet casein is used in the manufacture of Mozzarella cheese analogues. Hot solutions of calcium chelating salts are used to facilitate hydration of the protein which subsequently acts as an emulsifier for the oil phase dispersed in the protein/aqueous phase of the cheese analogue. On cooling, a matrix resembling cheese in texture and other rheological properties is produced. Considerable variations in the functional performance of different rennet caseins during cheese analogue manufacture have been observed. Commercial rennet caseins have been obtained and are being studied to establish the nature and causes of these variations in performance. A simplified model system to study rennet casein hydration was developed. The method involves dispersing rennet caseins in solutions of chelating salt at 55°C with stirring. The changes in viscosity index that occur as the rennet casein hydrated were followed using a controlled shear rate rheometer equipped with a custom designed paddle stirrer. Hydration of the rennet casein particles was found to occur in a number of stages including particle swelling, clumping, network formation and subsequent breakdown of the network. Considerable variations in the times taken for different rennet caseins to swell, clump and reach maximum viscosity index and in the stability of the hydrated protein dispersions were found. The rheological properties of the rennet caseins during hydration were highly dependent on the concentration of the chelating salt in the hydrating solution.     

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.     

The effect of storage temperatures and packaging methods on properties of Motal cheese

The effects of storage temperature (+4°C and -18°C) and packaging method (nonvacuum and vacuum) on biogenic amines in Motal cheese during storage periods were investigated. In addition, dry matter, titratable acidity, total nitrogen, water-soluble nitrogen, trichloroacetic acid-soluble nitrogen, phosphotungstic acid-soluble nitrogen, free amino group (proteolysis), electrophoretic patterns of casein, and amounts of lactic acid bacteria and coliforms were determined. Storage period had a significant effect on all of the biogenic amines. When compared with vacuum packaging, normal packaging had higher amounts of putrescine, cadaverine, histamine, and tyramine. Coliforms were not found at detectable levels (<100 cfu/g) in all cheese samples. Results of urea-PAGE analysis of cheese samples were in good agreement with biogenic amine results and other proteolysis parameters.     

Determinations of nonprotein nitrogen content of cheese whey.

Nonprotein nitrogen in pooled sweet (Cheddar and Dagano) and acid (cottage) wheys was estimated by currently proposed dialysis and chemical precipitation methods. Kjeldahl nitrogen analyses of (a) membrane retentates after water dialysis, and (b) 12% trichloroacetic acid plus .2% phosphotungstic acid filtrates indicated that nonprotein nitrogen values vary significantly with the method of sample preparation. Membrane porosity influence nonprotein nitrogen values for molecular weight cut-offs of 3500, 6000 to 8000 and 12,000 to 14,000. Dialyzable nitrogen values with all membranes were lower than 12% trichloroacetic acid soluble nitrogen for both wheys and higher than the 12% trichloroacetic acid + .2% phosphotungstic acid soluble nitrogen for all but the 3500 molecular weight cut-off membrane. The dialyzable nonprotein nitrogen fraction was heterogeneous but more that 80% was less than 3500 in molecular weight.     

The effect of two types of mould inoculants on the microbiological composition,physicochemical properties and protein hydrolysis in two Gorgonzola‐type cheese varieties during ripening

The aim of this paper was to investigate the role of two types of Penicillium roqueforti moulds (type esportazione and dolce) in the ripening of two Gorgonzola‐type cheese varieties. Cheeses were analysed after 4, 14, 30 and 60 days of ripening. Microbiological analysis showed high numbers of total bacterial count, yeasts and moulds in both 60‐day‐old cheese varieties. The concentration of water‐soluble N, nonprotein N and 5% phosphotungstic acid‐soluble N increased significantly during ripening. Patterns of proteolysis by urea‐polyacrylamide gel electrophoresis showed that rind‐to‐core gradients and age‐related changes in moisture and salt content influenced mould and other enzyme activities, which are reflected in various rates of protein degradation. The hydrolysis of αs₁‐ and β‐caseins was more extensive in the core than under the rind of both cheese varieties.     

Maillard reaction contributes to variability in hydration characteristics of rennet caseins in solutions of a calcium-sequestering salt

The objective of this study was to assess the effect of protein modification by Maillard reactions on the functionality of rennet caseins. The extent of Maillard reactions in rennet caseins was determined by measuring furosine levels by a high-performance liquid chromatography method following acid hydrolysis of the casein. An approximately 20-fold range in furosine levels in commercially produced rennet caseins was observed. High furosine levels correlated significantly with increased time taken to reach maximum viscosity index and with decreased maximum viscosity index when the rennet caseins were dispersed in a solution of calcium-sequestering salt, which suggests that modification of casein by Maillard reaction contributes to the variability in the hydration behaviour of the rennet caseins.     

Proteolysis,texture and colour of a raw goat milk cheese throughout the maturation

Changes on chemical and textural parameters were studied throughout maturation of PDO Ibores cheese. NCN/TN (non-casein nitrogen/total nitrogen) values were significantly modified (P < 0.001) throughout maturation, and significantly, lower values were observed at day 1 than at day 30, 60 and 90. Non-protein nitrogen/total nitrogen (NPN/TN) was not significantly changed during the ripening process (P > 0.05). Therefore, proteolysis extent (casein degradation and ‘ripening depth’) took place at initial stage and was limited throughout Ibores cheese maturation. In addition, since casein nitrogen decreased during the first month of ripening without a simultaneous increase in NPN/TN, this likely indicates that the large fragments of caseins liberated during this period are not further degraded into smaller products. Primary proteolysis (resulting from the action of endoproteases on caseins) is thus far more important than secondary proteolysis during this period. Moreover, secondary proteolysis remains limited along the 90 days. Polypeptide nitrogen significantly increased at day 30 from casein degradation compared to day 1, while free amino acids (FAA) content significantly increased during maturation process. In addition, hardness and adhesiveness values significantly increased, but cohesiveness and springiness significantly decreased up to day 60. Variables such as dry matter, NCN/TN and polypeptide nitrogen showed high correlations with textural parameters. Principal component analysis (PCA) of variables divided the Ibores cheeses according to their ripening: early, middle or late ripening.     

Proteolysis in rennet casein-based cheese analogues

Rennet caseins manufactured from mid- or late-lactation milk were used to prepare pilot- and commercial-scale Mozzarella-type cheese analogues. Mid-lactation rennet casein and analogues made from it contained low levels of plasmin; late-lactation rennet caseins and analogues made from them had higher levels of plasmin while pilot-scale analogues had higher levels of plasmin than commercial-scale analogues. The cheese analogues were stored at 8°C for 32 weeks during which time total bacterial counts increased from ∼10⁴ to 10⁷–10⁸ cfu g⁻¹, with non-starter lactic acid bacteria dominating the microflora. Proteolysis, as reflected by pH 4.6-soluble nitrogen and free amino acids, increased on storage of all the analogues. The increase was greater in the analogues manufactured from late-lactation rennet caseins than in those manufactured from mid-lactation rennet casein and in pilot-scale analogues than in commercial-scale analogues, reflecting the differences in plasmin levels as well as slight differences in composition. Polyacrylamide gel electrophoretograms of the cheese analogues showed that β-casein was hydrolysed more extensively than α_s1-casein and that the concentration of γ-caseins increased on storage. The degree of hydrolysis of β-casein was greater in the cheese analogues manufactured from late-lactation rennet caseins than in those manufactured from mid-lactation rennet casein and in the pilot-scale analogues than in the commercial-scale analogues reflecting the corresponding plasmin levels. Seventeen peptides in the pH 4.6-soluble extract of the cheeses were isolated and partially sequenced; all originated from the N-terminal region of β-casein; 9 peptides had Lys₂₉ of β-casein as the N-terminal, 6 peptides had Arg₁ of β-caesin as the N-terminal and the remaining two peptides had Ile₃₀ and Phe₃₃ of β-casein as the N-terminal, respectively. The pattern of casein hydrolysis in the cheese analogues suggests that plasmin was the primary proteolytic agent contributing to initial hydrolysis of the caseins while microbial proteinases and peptidases may have contributed to the high levels of free amino acids.     

Effect of commercial adjunct cultures on proteolysis in low-fat Kefalograviera-type cheese

The effect of two commercially available adjunct cultures, LBC 80 (Lactobacillus casei subsp. rhamnosus) and CR-213 (containing Lactococcus lactis subsp. cremoris and Lc. lactis subsp. lactis) on the proteolysis in low-fat hard ewes’ milk cheese of Kefalograviera-type was investigated. Two controls, a full-fat cheese (306 g kg⁻¹ fat, 378 g kg⁻¹ moisture) and a low-fat cheese (97 g kg⁻¹ fat, 486 g kg⁻¹ moisture, made using a modified procedure), were also prepared. The effect of adjunct culture on proteolysis, as examined by polyacrylamide gel electrophoresis of cheese and water soluble cheese extracts, was marginal. The reverse-phase HPLC peptide profiles of the water soluble extracts from low-fat cheeses were similar although some quantitative differences were observed between low-fat control cheese and experimental cheeses. The fat content as reflected by the differences in peptide profiles affected the pattern of proteolysis. Proteolysis, as measured by the percentage of total nitrogen soluble in water or in 120 g L⁻¹ trichloroacetic acid, was significantly (P<0.05) affected by the addition of adjunct cultures. Furthermore, the adjunct cultures enhanced the production of low molecular mass nitrogenous compounds; the levels of total nitrogen, soluble in 50 g L⁻¹ phosphotungstic acid, and of free amino acids were significantly (P<0.05) higher in the low-fat experimental cheeses than in the low-fat control cheese.     

Characteristics of Miniature Cheddar‐Type Cheese Made by Microbial Rennet from Bacillus amyloliquefaciens: A Comparison with Commercial Calf Rennet

Miniature Cheddar‐type cheeses were produced using microbial rennet from Bacillus amyloliquefaciens (milk‐clotting enzyme [MCE]) or calf rennet (CAR). With the exception of pH, there were no significant differences in gross composition between MCE‐cheese (MCE‐C) and CAR‐cheese (CAR‐C). The pH value of CAR‐C was significantly higher than that of MCE‐C at 40 and 60 d of ripening. The total nitrogen content of the pH 4.6‐soluble fraction obtained from MCE‐C was higher than that obtained from CAR‐C. However, nitrogen content of the 12% TCA‐soluble fraction was similar between CAR‐C and MCE‐C. The extent of α_s1‐casein and β‐casein hydrolysis, measured by urea‐PAGE, was similar in both cheese samples. The hydrolysis of β‐casein was lower than that of α_s1‐casein. Different reverse phase‐high‐performance liquid chromatography peptide profiles of ethanol‐soluble and ethanol‐insoluble fractions were obtained from CAR‐C and MCE‐C. The peptide content in the 2 cheese samples increased throughout ripening; the ratio of hydrophobic to hydrophilic peptides was lower in MCE‐C than in CAR‐C. Compared with CAR‐C, MCE‐C was softer as a result of higher protein hydrolysis. Microbial rennet from B. amyloliquefaciens contributed to higher proteolytic rates, which reduced ripening time.     

Protein profiles and total antioxidant capacity of water‐soluble and water‐insoluble fractions of white brined goat cheese at different stages of ripening

This study deals with proteolysis and total antioxidant capacity of proteins of white brined cheese prepared from overheated goat milk and ripened for fifty days. Proteolytic changes were reflected through the relatively low level of soluble nitrogen (50 days ripened cheese had 15.32 g/100 g of water‐soluble nitrogen, 8.1 g/100 g of TCA‐soluble nitrogen and 2.69 g/100 g of PTA‐SN), intensive proteolysis of α_s2‐CN during initial 10 days of ripening (up to 50.70% of initial content) and its much slower degradation through further 40 days, slow but continual decrease of β‐CN content (up to 85.14% of residual content) and high level of proteolytic products tightly bounded into gel network. Total antioxidant capacity of water‐soluble and water‐insoluble fractions increased after cheese ripening. These findings could be useful for better understanding and control over the production of white brined goat cheese as highly valuable functional product.