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.     

The effect of using an exopolysaccharide‐producing culture on the physicochemical properties of low‐fat and reduced‐fat Kasar cheeses

A mixed starter culture containing exopolysaccharide (EPS)‐producing strains of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus was combined with Lactobacillus helveticus LH301 and used in the manufacture of low‐fat and reduced‐fat Kasar cheeses. For comparison, low‐fat (C10) and reduced‐fat (C20) cheeses were made using EPS‐producing (EPS⁺) starter strain and EPS‐non‐producing (EPS^?) starter strain. The physicochemical properties of the cheeses were assessed in terms of chemical composition, texture, microstructure and microbial content over 90 days. Cheeses made with EPS‐producing culture (EPS10 and EPS20) had lower protein contents than control cheeses with 10% and 20% fat in dry basis (C10 and C20). Scanning electron microscopy images showed that using EPS‐producing culture resulted in a less compact protein matrix and sponge‐like structure in the cheese samples. In general, cheeses made using EPS‐producing culture had lower total viable counts. This could be related to the reduced survivability of EPS‐producing cells in the cheese matrix during ripening due to autolysis ability.     

Study of macromolecular interactions in low‐fat brined cheese modified with Zedu gum

The functionality of Zedu gum as a fat mimetic in low‐fat brined cheese was studied. The physicochemical, textural, rheological, microstructural and sensory properties of cheese samples modified with 0.1% and 0.25% of Zedu gum were compared to those of control cheeses (low‐fat and full‐fat cheeses with no fat mimetic) during ripening. To obtain further information about the cheeses' structure and interactions between macromolecules (casein protein and Zedu gum), other parameters were analysed by differential scanning calorimetry and Fourier transform infrared (FTIR) spectroscopy. Incorporation of Zedu gum into low‐fat cheese caused an open microstructure and softer texture in comparison with the control low‐fat cheese. The thermal properties and FTIR spectra of the cheeses were influenced by both fat mimetic and ripening time. On days 1 and 60 of ripening time, the lower value of enthalpy of the low‐fat cheese with 0.25 g of Zedu gum/kg of milk (AS 0.25) in comparison with control low‐fat cheese could have been due to the electrostatic nature of the interactions between Zedu gum and casein protein. On both days, the FTIR spectrum of AS 0.25 showed a well separated absorption at 1746 cm^?1 possibly due to the formation of ester groups as a result of the interaction of the carbonyl groups in Zedu gum with the hydroxyl groups of some amino acids in casein.     

Proteolytic properties of Turkish white‐brined cheese (Beyaz peynir) made by using wild‐type Lactococcal strains

The development of proteolysis in white‐brined Turkish cheese made by using wild strains of Lactococcus lactis subsp. lactis (namely MBLL9, MBLL23 and MBL27) was monitored for 90 days. Proteolysis in cheeses was investigated using urea‐PAGE gel electrophoresis of pH 4.6‐insoluble and RP‐HPLC of both 70% ethanol‐insoluble and 70% ethanol‐soluble nitrogen fractions. Results indicated that developments of proteolysis in the experimental cheeses were strain dependent. The degradation of casein fractions was more evident in the cheeses made using strain MBLL23. The lowest levels of proteolysis and development of acidity were obtained in the cheese made using strain MBLL9.     

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.     

Angiotensin‐converting enzyme‐inhibitory and antithrombotic activities of soluble peptide extracts from buffalo and cow milk Cheddar cheeses

The aim of the study was to evaluate potential role of a water‐soluble peptide (WSP) extracts derived from buffalo and cow milk Cheddar cheeses with special reference to their antihypertension and antithrombotic activities. The WSP fractions collected at different stages of ripening were tested to assess their degree of proteolysis, their peptides were profiled by RP‐HPLC and in vitro assays for potential bioactivity were conducted. The peptide peak development was observed with slight differences in peaks number, area and height. Both angiotensin‐converting enzyme‐inhibitory and antithrombotic activities increased progressively during ripening. In comparison, the highest activities were observed in peptide extracts obtained from buffalo milk Cheddar cheese, in a dose‐dependent fashion.     

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.     

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.     

The effects of starter cultures on chemical,biochemical and sensory properties of low‐fat Tulum cheeses during ripening

Three different commercial starter cultures, Choozit^? MA 11 (MA ), Choozit^? BT 01 (BT ) and Choozit^? Feta A (Feta), were used to remedy textural and aromatic defects and improve the overall quality of low‐fat Tulum cheeses. Chemical and sensory analyses as well as electrophoresis were performed. Supplemental yoghurt bacteria and Lactobacillus helveticus were found to be key contributors in proteolysis with varying protein breakdown capacities. The results suggest that using appropriate culture combinations could result in low‐fat Tulum cheeses with better sensory characteristics and proteolysis rates.     

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

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

Ultrafiltered milk reduces bitterness in reduced-fat Cheddar cheese made with an exopolysaccharide-producing culture

The objectives were to reduce bitterness in reduced-fat Cheddar cheese made with an exopolysaccharide (EPS)-producing culture and study relationships among ultra-filtration (UF), residual chymosin activity (RCA), and cheese bitterness. In previous studies, EPS-producing cultures improved the textural, melting, and viscoelastic properties of reduced-fat Cheddar cheese. However, the EPS-positive cheese developed bitterness after 2 to 3 mo of ripening due to increased RCA. We hypothesized that the reduced amount of chymosin needed to coagulate UF milk might result in reduced RCA and bitterness in cheese. Reduced-fat Cheddar cheeses were manufactured with EPS-producing and nonproducing cultures using skim milk or UF milk (1.2×) adjusted to a casein:fat ratio of 1.35. The EPS-producing culture increased moisture and RCA in reduced-fat Cheddar cheese. Lower RCA was found in cheese made from UF milk compared with that in cheese made from control milk. Ultrafiltration at a low concentration rate (1.2×) produced EPS-positive, reduced-fat cheese with similar RCA to that in the EPS-negative cheese. Slower proteolysis was observed in UF cheeses compared with non-UF cheeses. Panelists reported that UF EPS-positive cheese was less bitter than EPS-positive cheese made from control milk. This study showed that UF at a low concentration factor (1.2×) could successfully reduce bitterness in cheese containing a high moisture level. Because this technology reduced the RCA level (per g of protein) to a level similar to that in the control cheeses, the contribution of chymosin to cheese proteolysis would be similar in both cheeses.     

Changes during ripening in chemical composition,proteolysis, volatile composition and texture in Kashar cheese made using raw bovine,ovine or caprine milk

Kashar cheeses were manufactured from pure ovine (OV), bovine (BV) and caprine (CP) milk, and the chemical composition, cheese yield, proteolysis, hardness, meltability and volatile composition were studied during 90 days. Gross chemical composition, cheese yield and level of proteolysis were higher in OV cheeses than those of BV or CP cheeses. Glu, Val, Leu, Phe and Lys were the most abundant free amino acids (FAA) in the samples, and the concentrations of individual FAA were at the highest levels in OV cheeses with following BV and CP cheeses. Urea‐PAGE patterns and RP‐HPLC peptide profiles of the BV cheeses were completely different from the small ruminants’ milk cheeses (OV or CP). Higher and lower hardness and meltability values were observed in CP cheeses, respectively. OV cheeses resulted in higher levels of the major volatile compounds. In conclusion, the Kashar cheese made using OV milk can be recommended due to high meltability, proteolysis and volatiles.     

Application of exopolysaccharide-producing cultures in reduced-fat Cheddar cheese: texture and melting properties

Textural, melting, and sensory characteristics of reduced-fat Cheddar cheeses made with exopolysaccharide (EPS)-producing and nonproducing cultures were monitored during ripening. Hardness, gumminess, springiness, and chewiness significantly increased in the cheeses as fat content decreased. Cheese made with EPS-producing cultures was the least affected by fat reduction. No differences in hardness, springiness, and chewiness were found between young reduced fat cheese made with a ropy Lactococcus lactis ssp. cremoris [JFR1; the culture that produced reduced-fat cheese with moisture in the nonfat substance (MNFS) similar to that in its full-fat counterpart] and its full-fat counterpart. Whereas hardness of full-fat cheese and reduced-fat cheese made with JFR1 increased during ripening, a significant decrease in its value was observed in all other cheeses. After 6 mo of ripening, reduced fat cheeses made with all EPS-producing cultures maintained lower values of all texture profile analysis parameters than did those made with no EPS. Fat reduction decreased cheese meltability. However, no differences in meltability were found between the young full-fat cheese and the reduced-fat cheese made with the ropy culture JFR1. Both the aged full- and reduced-fat cheeses made with JFR1 had similar melting patterns. When heated, they both became soft and creamy without losing shape, whereas reduced-fat cheese made with no EPS ran and separated into greasy solids and liquid. No differences were detected by panelists between the textures of the full-fat cheese and reduced-fat cheese made with JFR1, both of which were less rubbery or firm, curdy, and crumbly than all other reduced-fat cheeses.     

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.     

Proteolysis on Reggianito Argentino cheeses manufactured with natural whey cultures and selected strains of Lactobacillus helveticus

Reggianito Argentino cheese is traditionally manufactured with whey starter cultures that provide typical and intense flavor but can cause poor quality standardization. In this study, the influence of natural and selected starters on Reggianito Argentino cheese proteolysis was investigated. Cheeses were manufactured with three strains of Lactobacillus helveticus (SF133, SF138 and SF209) cultured individually in sterile whey and used as single or mixed starters. Control cheeses were made with natural whey starter culture. Cheeses were analyzed to determine gross composition, as well as total thermophilic lactic flora. Proteolysis was assessed by N fractions, electrophoresis and liquid chromatography. Gross composition of the cheeses did not significantly differ, while viable starter cell counts were lower for cheeses made with strain SF209 alone or combined with other strains. Soluble N at pH 4.6 was the same for cheeses made with natural or selected starters, but soluble N in 12% trichloroacetic acid and 2.5% phosphotungstic acid was significantly higher in cheeses made with starters containing strain SF209. Nitrogen fractions results indicated that natural whey starter cultures could be replaced by several starters composed of the selected strains without significant changes to proteolysis patterns. Starter cultures prepared only with SF209 or with the three selected L. helveticus strains produced cheese products with significantly more proteolysis than control cheeses. Chromatographic profiles analyzed by principal components showed that three main peaks on chromatograms, presumptively identified as Tyr, Phe, and Trp, explained most of variability. Principal component scores indicated that cheese samples were grouped by ripening time, which was confirmed by linear discriminant analysis. On the contrary, samples did not cluster by Lactobacillus strain or type of starter.     

Primary and Secondary Proteolysis in Eleven Turkish Cheese Varieties

Levels of proteolysis of 75 samples belonging to 11 Turkish cheese varieties, including Civil, Canak, Dil, Divle Tulum, Ezine, Hellim, Malatya, Mihalic, Orgu, Urfa, and Van Otlu, were comparatively studied. The cheeses were mainly produced using traditional methods; however, some varieties were industrially produced. Chemical composition and the levels of soluble nitrogen fractions of the cheeses varied depending on the cheese variety. Gel electrophoresis of the cheeses showed that the samples presented different gel patterns with α_s1-casein being extensively degraded in many cheeses; whereas the hydrolysis of α_s1-casein in Malatya and Hellim was observed to be limited. Peptide profiles by RP-HPLC of the water-soluble fractions were largely different for many of the samples, but some similarities were visualized. Multivariate analysis of the RP-HPLC data grouped the cheeses according to their peptide profiles. The results suggested that each variety of cheese had different levels of proteolysis. The manufacturing technique and ripening conditions employed have played a determinative role on the proteolytic patterns of the cheeses analyzed.     

Utilization of interesterified fat in the production of Turkish white cheese

The chemical, physicochemical, proteolysis, sensory, and texture characteristics of white cheeses made from interesterified fat were examined throughout ripening for 90 days. The water-soluble nitrogen based ripening indexes of cheeses increased throughout the ripening period. However, there were not large quantitative differences between the peptide profiles of the all cheese samples. Cheeses produced by using fully interesterified fat had higher values for hardness, chewiness, and gumminess than that of control cheese (p<0.05). The polyunsaturated to saturated fatty acid ratios of cheeses were increased due to the presence of interesterified fat. The cholesterol values of cheeses decreased at the rate of between 58.83–89.04% depending on interesterified fat addition. In the sensory analysis, similar scores were obtained for both the control cheese and the other cheeses. The results showed that interesterified fat in cheese production could be used to fully or partially replace the milk fat in cheese.     

Lipolysis and proteolysis profiles of fresh artisanal goat cheese made with raw milk with 3 different fat contents

The objective of this study was to describe the proteolysis and lipolysis profiles in goat cheese made in the Canary Islands (Spain) using raw milk with 3 different fat contents (0.5, 1.5, and 5%) and ripened for 1, 7, 14, and 28 d. β-Casein was the most abundant protein in all cheeses and at all ripening times. Quantitative analysis showed a general decrease in caseins as ripening progressed, and degradation rates were higher for α_S1-casein than for β-casein and α_S2-casein. Furthermore, the degradation rate during the experimental time decreased with lower fat contents. The α_S2-casein and α_S1-casein levels that remained in full-fat and reduced-fat cheeses were less than those in low-fat cheese. In contrast, β-casein also showed degradation along with ripening, but differences in degradation among the 3 cheese types were not significant at 28 d. The degradation products increased with the ripening time in all cheeses, but they were higher in full-fat cheese than in reduced-fat and low-fat cheeses. The free fatty acid concentration per 100 g of cheese was higher in full-fat cheese than in reduced- and low-fat cheese; however, when the results were expressed as milligrams of free fatty acids per gram of fat in cheese, then lipolysis occurred more rapidly in low-fat cheese than in reduced- and full-fat cheeses. These results may explain the atypical texture and off-flavors found in low-fat goat cheeses, likely the main causes of non-acceptance.     

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.