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.     

Microbial, chemical and sensorial variables of the Spanish traditional blue-veined Cabrales cheese, as affected by inoculation with commercial Penicillium roqueforti spores

This paper reports the technological effects of inoculating Cabrales cheese (a traditional, Spanish, blue-veined cheese) with Penicillium roqueforti spores. Three batches of inoculated Cabrales cheese were manufactured and a number of their microbial and biochemical variables recorded. The results were compared with those obtained for three batches of control cheese made using traditional technology (i.e., adding neither starter cultures nor fungal spores). Although mould and yeast populations grew more quickly in the inoculated cheeses, their normally dominant and representative microbial populations were not affected neither was their gross biochemical composition changed. The variations observed were thought to be caused by the uncontrolled environmental conditions of manufacture and ripening. The development of free amino acids and volatile compounds was significantly increased at 30 days in the inoculated cheeses, although the values for both types of cheese were almost identical at 90 days. The inoculated cheeses obtained higher scores in a hedonistic sensorial evaluation. Thus, inoculation improved the standardization and quality of the cheese.     

Effect of some technological parameters on microbiological, chemical and sensory qualities of Civil cheese during ripening

The objective of this research was to determine the effects of different ripening methods [brine salting, dry salting, incorporating with Lor cheese (LR) and vacuum packaging] of Civil cheeses on its microbiological, chemical and sensory properties. Civil cheeses were analysed on the 2nd, 30th, 60th and 90th day of ripening. Chemical compositions of the cheeses were significantly different. While the highest dry matter and titratable acidity values were determined on dry salted cheeses, the highest fat and fat in dry matter contents were found in Civil cheese ripened together with LR. The water-soluble nitrogen and ripening index values were lower in cheese ripened incorporating with LR. Excessive proteolysis was not seen in any of cheese samples. The ripening in different methods affected microbiological and sensory properties of Civil cheese. Panellists preferred vacuum packaging and dry-salting cheeses compared to the other samples on the 90th day of 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.     

Influence of exopolysaccharide‐producing cultures on the volatile profile and sensory quality of low‐fat Tulum cheese during ripening

The objective of this investigation was to compare the composition and changes in the concentration of volatiles in low‐fat and full‐fat Tulum cheeses during ripening. Tulum cheese was manufactured from low‐ or full‐fat milk using exopolysaccharide (EPS)‐producing or non‐EPS‐producing starter cultures. A total of 82 volatile compounds were identified belonging to the following chemical groups: acids (seven), esters (21), ketones (14), aldehydes (six), alcohols (14) and miscellaneous compounds (20). The relative amounts of acids, alcohols and aldehydes increased in the cheeses made with EPS‐producing cultures during 90 days of ripening. Differences were found in the volatile profile of full‐fat Tulum cheese compared with the low‐fat variant, especially after 90 days of ripening. Exopolysaccharide‐producing cultures changed the volatile profile, and the EPS‐producing cultures including Streptococcus thermophilus + Lactobacillus delbrueckii subsp. bulgaricus + Lactobacillus helveticus (LF‐EPS2) produced cheese with higher levels of methyl ketones and aldehydes than the non‐EPS cultures. In the sensory analysis, full‐fat Tulum cheeses and the cheese produced with the EPS‐producing culture containing Lb. helveticus (LF‐EPS2) were preferred by the expert panel. It was concluded that the use of EPS‐producing starter cultures in the manufacture of low‐fat Tulum cheese had the potential to improve the flavour.     

Survival of micro‐organisms and organic acid profile of probiotic Cheddar cheese from buffalo milk during accelerated ripening

The study aimed to assess the impact of ripening at elevated temperatures on the survival of probiotic micro‐organisms and production of organic acids in Cheddar cheese. Cheese was manufactured from buffalo milk using lactococci starters along with different probiotic bacteria (Lactobacillus acidophilus LA‐5, Bifidobacterium bifidum Bb‐11 and Bifidobacterium longum BB536) as adjunct cultures. The cheeses were ripened at 4–6 °C or 12–14 °C for 180 days and examined for composition, organic acids and microbial survival. The production of organic acids was accelerated at 12–14 °C when compared to normal ripening temperatures. The probiotic bacteria increased production of lactic and acetic acids, compared to cheese made with lactococci alone. The survival of the mesophilic starters was significantly (P < 0.05) reduced in all the cheese samples ripened at the higher temperature. However, the probiotic bacteria remained viable (>7.0 log₁₀ cfu/g) throughout the 180 days of ripening, irrespective of temperature. It was concluded that Cheddar containing additional probiotic cultures can effectively be ripened at elevated temperatures without any adverse effects.     

The effects of some starter cultures on the properties of Turkish White cheese

White cheese samples were manufactured from bovine milk using three different commercial direct vat starter cultures (DVS-1, -2 and -3) and a lyophilized culture, and ripened at 4 ± 1°C for 90 days. The composition, titratable acidity and ripening indices of the cheese samples were determined on the 2nd, 30th, 60th and 90th days of ripening. The ratios of total solids, protein and fat were higher for cheeses manufactured using DVS-2 and lyophilized cultures but the titratable acidity in cheese produced using DVS-3 and lyophilized cultures was higher (P < 0.01). The mean value of the ripening indices of the cheese produced using the lyophilized culture was lower than the cheeses produced with added DVS cultures (P < 0.05). The total solids, ash, salt ratios, titratable acidity and ripening indices values increased for all types of white cheeses during ripening (P < 0.05).     

Proteolysis texture and microstructure of low‐fat Tulum cheese affected by exopolysaccharide‐producing cultures during ripening

The objective of the study was to determine the effects of exopolysaccharide (EPS)‐producing or non‐EPS‐producing starters on proteolysis, physical and microstructural characteristics of full‐fat or low‐fat Tulum cheeses during ripening. For this purpose, Tulum cheese was manufactured using full‐ or low‐fat milk with EPS‐producing and non‐EPS‐producing starter cultures. Chemical composition, proteolysis, texture profiles and microstructure of the cheeses were studied during 90 days of ripening. Urea‐PAGE of water‐insoluble and RP‐HPLC peptide profiles of water‐soluble fractions of the cheeses showed that the use of starters resulted in different degradation patterns in all cheeses during ripening. Although β‐casein exhibited similar degradation patterns in all cheeses, small differences are present in α_s1‐casein degradation during ripening. Reducing fat in Tulum cheese changed the RP‐HPLC peptide profile of the cheeses. The use of EPS‐producing cultures improved the textural characteristics and changed the microstructure and proteolysis of low‐fat Tulum cheese.     

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.     

The effects of using a starter culture,lipase, and protease enzymes on ripening of Mihalic cheese

The purpose of this study was to determine the effects of fungal lipase from Mucor miehei and a bacterial neutral protease from Bacillus subtilis alone and combined with a starter culture on ripening properties of traditional Turkish Mihalic cheese. The use of protease with lipase (Cult + Prot + Lip) resulted in better flavour and texture with accelerated ripening. The obtained results pointed out that the gross compositions of the cheeses were changed by the type of enzymes and ripening time (P < 0.01). The acid degree value (ADV) of all cheeses showed a linear increase with ripening. The highest lipolysis rate was noted in lipase‐added cheese batch (as 5.56 ADV) with highest γ‐CN ratio and β‐CN degradation. At the end of ripening time, it was observed that αs‐CN ratios decreased in starter‐added (Cult), starter + protease–added (Cult + Prot), and protease‐added (Prot) cheese batches. The use of protease with lipase (Cult + Prot + Lip) resulted in better flavour and texture with accelerated ripening. Protease‐added cheeses, which were characterized by bitterness and crumbly textural properties owing to the intense breakdown of β‐casein, scored lower than lipase‐added cheeses. It was determined that the use of mesophilic aromatic starter culture with lipase and protease could be used to accelerate ripening of Mihalic cheese made from pasteurised milk.     

Compositional,biochemical and textural changes during ripening of Tulum cheese made with different coagulants

In the present study, biochemical, chemical and texture changes in Tulum cheeses made using calf rennet and microbial rennets (Aspergillus niger protease and Rhizomucor miehei protease) were compared during ripening for up to 90 days. A total of 15 free fatty acids (FFAs) were detected in the cheese samples. The peroxide values (PV) of the cheeses increased significantly (P < 0.05) during ripening and the cheese made with calf rennet had the highest PV. Proteolysis in the cheeses increased as the ripening time increased. α_s1‐casein and β‐casein degradation was higher in cheeses manufactured with R. miehei protease. Cheeses made with calf rennet were significantly (P < 0.05) harder, more adhesive, more cohesive and more resilient than those made with microbial rennet.     

Effect of starter cultures on the physico‐chemical and biochemical features in ewe cheese made with extracts from flowers of Cynara cardunculus L

The effect of starter cultures on the physico‐chemical and biochemical features of Los Pedroches cheese made by using Cynara Cardunculus L extracts as vegetable coagulant was assessed. Specimens of cheese containing no starter (vats A and C) and others to which a lactic acid starter was added (vats B and D) were stored either under typical farmhouse ripening conditions (vats A and B) or in a controlled chamber at 14°C and 80% relative humidity (vats C and D) for 90 days. The addition of starter cultures and the ripening conditions where found to have no appreciable effect on the moisture, fat, protein, ash, NaCl, Ca and P contents of the cheese, nor on its water activity; however, it resulted in increased lactic acid contents and lower pH values relative to control specimens throughout ripening. The protein breakdown of the cheeses was assessed in terms of soluble nitrogen, non‐protein nitrogen, aminoacid nitrogen, ammonia nitrogen, soluble tyrosine and tryptophan, and urea‐PAGE. Proteolysis was generally more marked and rapid in cheese containing the lactic starter, the NPN, NH₃ ‐N and soluble Tyr and Trp contents of which were significantly higher than those of the cheese obtained with no starter. The PAGE technique revealed no appreciable differences between the four cheese batches studied; however, it showed the cheese to be more resistant to β‐CN hydrolysis than to α_s‐CN hydrolysis. In general, the flavour and aroma scores were higher in batches obtained with starter cultures and ripened under controlled conditions. © 1999 Society of Chemical Industry     

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.     

Use of principal component analysis to evaluate the physical properties of Mahon cheese

 The texture, colour and water activity of the seven existing types of Mahon cheese, manufactured under the Mahon cheese Appellation of Origin, were studied using eight physical variables. Four of the types are industrially manufactured [fresh (less than 10 days of ripening), half-ripened (2–5 months of ripening), ripened (5–10 months of ripening) and old-ripened (more than 10 months of ripening)] and three are traditionally manufactured (half-ripened, ripened and old-ripened). ANOVA, Tukey's multiple range test and principal component analysis (PCA) were used. Mahon cheeses from different ripening periods exhibited significant differences in water activity and yellowness values (P<0.001). PCA reduced the eight physical variables to two independent components, which accounted for 84.4% of the total variance. The first principal component (PC1) separated cheeses characterised by high water activity, high springiness values and slight yellowness from those with high hardness and puncture force; PC1 therefore distinguished among samples from different ripening periods. Cohesiveness, chewiness and gumminess were highly correlated in the second principal component, which could be interpreted as an index of cohesiveness. Received: 8 February 1999     

The effects of beeswax coating on quality of Kashar cheese during ripening

The aim of this study was to evaluate the effects of the application of beeswax coating on the microbiological, physicochemical and sensory properties of Kashar cheese during ripening (120 day). Kashar cheeses were coated with two different thickness of beeswax (single‐layer coating, BW1, and double‐layer coating, BW2). For comparison, vacuum packaged (VP) and without packaging material (control) were also studied. Generally, no differences were found in total aerobic mesophilic bacteria, LAB on M‐17 agar, coliform bacteria and S. aureus counts among cheeses. Microbiological analyses also showed the beeswax‐coated cheeses presented a decrease of 2.5 logarithmic units on mould counts compared to control at 120th day. The control cheese had significantly (P < 0.05) higher dry matter, fat and protein contents, followed by BW1. However, the coating reduced formation of a thick crust layer by delaying moisture loss. At the end of 120‐day storage period, no significant differences in pH and acidity values were observed among the cheeses studied. Compared to other cheeses, control and BW1 cheeses had higher levels of WSN and ripening index in the end of storage. In the result of sensory analysis, while cheese BW1 and control were more preferred by the panellists, cheese VP received the lowest scores.     

Improvement of texture and structure of reduced-fat Cheddar cheese by exopolysaccharide-producing lactococci

The objective of this study was to evaluate the effect of capsular and ropy exopolysaccharide (EPS)-producing strains of Lactococcus lactis ssp. cremoris on textural and microstructural attributes during ripening of 50%-reduced-fat Cheddar cheese. Cheeses were manufactured with added capsule- or ropy-forming strains individually or in combination. For comparison, reduced-fat cheese with or without lecithin added at 0.2% (wt/vol) to cheese milk and full-fat cheeses were made using EPS-nonproducing starter, and all cheeses were ripened at 7°C for 6 mo. Exopolysaccharide-producing strains increased cheese moisture retention by 3.6 to 4.8% and cheese yield by 0.28 to 1.19 kg/100 kg compared with control cheese, whereas lecithin-containing cheese retained 1.4% higher moisture and had 0.37 kg/100 kg higher yield over the control cheese. Texture profile analyses for 0-d-old cheeses revealed that cheeses with EPS-producing strains had less firm, springy, and cohesive texture but were more brittle than control cheeses. However, these effects became less pronounced after 6 mo of ripening. Using transmission electron microscopy, fresh and aged cheeses with added EPS-producing strains showed a less compact protein matrix through which larger whey pockets were dispersed compared with control cheese. The numerical analysis of transmission electron microscopy images showed that the area in the cheese matrix occupied by protein was smaller in cheeses with added EPS-producing strains than in control cheese. On the other hand, lecithin had little impact on both cheese texture and microstructure; after 6 mo, cheese containing lecithin showed a texture profile very close to that of control reduced-fat cheese. The protein-occupied area in the cheese matrix did not appear to be significantly affected by lecithin addition. Exopolysaccharide-producing strains could contribute to the modification of cheese texture and microstructure and thus modify the functional properties of reduced-fat Cheddar cheese.     

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     

Effects of different ripening procedures on the final characteristics of Picante cheese

 Picante da Beira Baixa (or Picante) cheese is a hard, piquant, salted traditional cheese manufactured in Portugal from raw sheep's and goat's milks. The purpose of this work was to quantitatively assess the influence of various ripening procedures on the final characteristics of Picante cheese. Two alternative ripening protocols were considered, the traditional one and another with controlled environmental conditions via use of maturation chambers set at different preselected temperatures. The experimental cheeses were characterised in terms of microbiological, physicochemical, biochemical, sensorial and textural properties. Ripening time and temperature were statistically significant parameters for all microflora. The two ripening methods led to statistically significant differences in all physicochemical and biochemical parameters, especially the moisture content and the soluble nitrogen fractions (i.e. water loss was slower and proteolysis was faster in cheeses ripened via the traditional method). Differences in microbiological, physicochemical and biochemical properties were probables implicated in differences in textural and sensorial properties, especially cheese hardness and flavour. It was concluded that the standard ripening method was closest to the traditional one in terms of final cheese characteristics when the ripening temperature was above 11.5  °C. Received: 3 February 1998     

Effects of different ripening procedures on the final characteristics of Picante cheese

 Picante da Beira Baixa (or Picante) cheese is a hard, piquant, salted traditional cheese manufactured in Portugal from raw sheep's and goat's milks. The purpose of this work was to quantitatively assess the influence of various ripening procedures on the final characteristics of Picante cheese. Two alternative ripening protocols were considered, the traditional one and another with controlled environmental conditions via use of maturation chambers set at different preselected temperatures. The experimental cheeses were characterised in terms of microbiological, physicochemical, biochemical, sensorial and textural properties. Ripening time and temperature were statistically significant parameters for all microflora. The two ripening methods led to statistically significant differences in all physicochemical and biochemical parameters, especially the moisture content and the soluble nitrogen fractions (i.e. water loss was slower and proteolysis was faster in cheeses ripened via the traditional method). Differences in microbiological, physicochemical and biochemical properties were probables implicated in differences in textural and sensorial properties, especially cheese hardness and flavour. It was concluded that the standard ripening method was closest to the traditional one in terms of final cheese characteristics when the ripening temperature was above 11.5  °C. Received: 3 February 1998     

Application of principal component analysis to chemical characteristics of Mahon cheese

 The chemical composition of the seven existing types of Mahon cheese, manufactured under the “Mahon” cheese Appellation of Origin, were studied. Of the cheeses, four were industrial (fresh, half-ripened, ripened and old-ripened) and three were traditionally manufactured (half-ripened, ripened and old-ripened). The chemical parameters were evaluated using ANOVA, Tukey’s multiple range test and principal component (PCA) analyses. Significant differences were found between Mahon cheeses ripened for different times in terms of moisture and non-protein nitrogen (NPN) contents (p <0.001); these differences were found to be independent of the manufacturing methodology, i.e. industrial or traditional. The ash and fat contents of industrially manufactured cheeses were significantly different from those in traditionally manufactured cheeses (p <0.001). PCA reduced chemical variables to two independent components: the first principal component was high in moisture, NPN content and water activity; the second, in NaCl, ash and fat contents. These results indicate that the seven types of Mahon cheese described by the “Mahon” cheese Appellation of Origin can be differentiated by their chemical characteristics. Received: 4 April 1997 / Revised version: 4 July 1997