Effect of coagulant type and level on the properties of half-salt,half-fat Cheddar cheese made with or without adjunct starter: Improving texture and functionality

The potential of increasing proteolysis as a means of enhancing the texture and heat-induced flow of half-fat, half-salt Cheddar cheese made with control culture (CL, Lactococcus lactis subsp. cremoris/lactis) or adjunct culture (AC, CL + Lactobacillus helveticus) was investigated. Proteolysis was altered by substituting bovine chymosin (BC) with camel chymosin (CC), or by a 2.5-fold increase in level of BC. In cheese with CL-culture, increasing BC led to a large increase in pH and more rapid degradation of α_S1-casein during maturation, and cheese that was less firm after 180 d. In contrast, substitution of BC with CC in cheeses made with CL-culture had an opposite effect. While chymosin type and level had a similar influence on α_S1-casein hydrolysis in the AC-culture cheeses, it did not affect texture or flowability. Grading indicated that cheese made with AC-culture and with a higher level of BC was the most appealing.     

Influence of starters on chemical, biochemical, and sensory changes in Turkish White-brined cheese during ripening

Turkish White-brined cheese was manufactured using Lactococcus strains (Lactococcus lactis ssp. lactis NCDO763 plus L. lactis ssp. cremoris SK11 and L. lactis ssp. lactis UC317 plus L. lactis ssp. cremoris HP) or without a starter culture, and ripened for 90 d. It was found that the use of starters significantly influenced the physical, chemical, biochemical, and sensory properties of the cheeses. Chemical composition, pH, and sensory properties of cheeses made with starter were not affected by the different starter bacteria. The levels of soluble nitrogen fractions and urea-PAGE of the pH 4.6-insoluble fractions were found to be significantly different at various stages of ripening. Urea-PAGE patterns of the pH 4.6-insoluble fractions of the cheeses showed that considerable degradation of α_s1-casein occurred and that β-casein was more resistant to hydrolysis. The use of a starter culture significantly influenced the levels of 12% trichloroacetic acid-soluble nitrogen, 5% phosphotungstic acid-soluble nitrogen, free amino acids, total free fatty acids, and the peptide profiles (reverse phase-HPLC) of 70% (vol/vol) ethanol-soluble and insoluble fractions of the pH 4.6-soluble fraction of the cheeses. The levels of peptides in the cheeses increased during the ripening period. Principal component and hierarchical cluster analyses of electrophoretic and chromatographic results indicated that the cheeses were significantly different in terms of their peptide profiles and they were grouped based on the use and type of starter and stage of ripening. Levels of free amino acid in the cheeses differed; Leu, Glu, Phe, Lys, and Val were the most abundant amino acids. Nitrogen fractions, total free amino acids, total free fatty acids, and the levels of peptides resolved by reverse phase-HPLC increased during ripening. No significant differences were found between the sensory properties of cheeses made using a starter, but the cheese made without starter received lower scores than the cheeses made using a starter. It was found that the cheese made with strains NCDO763 plus SK11 had the best quality during ripening. It was concluded that the use of different starter bacteria caused significant differences in the quality of the cheese, and that each starter culture contributed to proteolysis to a different degree.     

Genotypic identification of some lactic acid bacteria by amplified fragment length polymorphism analysis and investigation of their potential usage as starter culture combinations in Beyaz cheese manufacture

In this study, 2 different starter culture combinations were prepared for cheesemaking. Starter culture combinations were formed from 8 strains of lactic acid bacteria. They were identified as Lactococcus lactis ssp. lactis (2 strains), Lactobacillus plantarum (5 strains), and Lactobacillus paraplantarum (1 strain) by amplified fragment length polymorphism analysis. The effects of these combinations on the physicochemical and microbiological properties of Beyaz cheeses were investigated. These cheeses were compared with Beyaz cheeses that were produced with a commercial starter culture containing Lc. lactis ssp. lactis and Lc. lactis ssp. cremoris as control. All cheeses were ripened in brine at 4°C for 90 d. Dry matter, fat in dry matter, titratable acidity, pH, salt in dry matter, total N, water-soluble N, and ripening index were determined. Sodium dodecyl sulfate-PAGE patterns of cheeses showed that α_S-casein and β-casein degraded slightly during the ripening period. Lactic acid bacteria, total mesophilic aerobic bacteria, yeast, molds, and coliforms were also counted. All analyses were repeated twice during d 7, 30, 60, and 90. The starter culture combinations were found to be significantly different from the control group in pH, salt content, and lactobacilli, lactococci, and total mesophilic aerobic bacteria counts, whereas the cheeses were similar in fat, dry matter content, and coliform, yeast, and mold counts. The sensory analysis of cheeses indicated that textural properties of control cheeses presented somewhat lower scores than those of the test groups. The panelists preferred the tastes of treatment cheeses, whereas cheeses with starter culture combinations and control cheeses had similar scores for appearance and flavor. These results indicated that both starter culture combinations are suitable for Beyaz cheese production.     

Biochemical patterns in ovine cheese: Influence of probiotic strains

This study was undertaken to evaluate the effect of lamb rennet paste containing probiotic strains on proteolysis, lipolysis, and glycolysis of ovine cheese manufactured with starter cultures. Cheeses included control cheese made with rennet paste, cheese made with rennet paste containing Lactobacillus acidophilus culture (LA-5), and cheese made with rennet paste containing a mix of Bifidobacterium lactis (BB-12) and Bifidobacterium longum (BB-46). Cheeses were sampled at 1, 7, 15, and 30 d of ripening. Starter cultures coupled with probiotics strains contained in rennet paste affected the acidification and coagulation phases leading to the lowest pH in curd and cheese containing probiotics during ripening. As consequence, maturing cheese profiles were different among cheese treatments. Cheeses produced using rennet paste containing probiotics displayed higher percentages of α_S1-I-casein fraction than traditional cheese up to 15 d of ripening. This result could be an outcome of the greater hydrolysis of α-casein fraction, attributed to higher activity of the residual chymosin. Further evidence for this trend is available in chromatograms of water-soluble nitrogen fractions, which indicated a more complex profile in cheeses made using lamb paste containing probiotics versus traditional cheese. Differences can be observed for the peaks eluted in the highly hydrophobic zone being higher in cheeses containing probiotics. The proteolytic activity of probiotic bacteria led to increased accumulation of free amino acids. Their concentrations in cheese made with rennet paste containing Lb. acidophilus culture and cheese made with rennet paste containing a mix of B. lactis and B. longum were approximately 2.5 and 3.0 times higher, respectively, than in traditional cheese. Principal component analysis showed a more intense lipolysis in terms of both free fatty acids and conjugated linoleic acid content in probiotic cheeses; in particular, the lipolytic pattern of cheeses containing Lb. acidophilus is distinguished from the other cheeses on the basis of highest content of health-promoting molecules. The metabolic activity of the cheese microflora was also monitored by measuring acetic, lactic, and citric acids during cheese ripening. Cheese acceptability was expressed for color, smell, taste, and texture perceived during cheese consumption. Use of probiotics in trial cheeses did not adversely affect preference or acceptability; in fact, panelists scored probiotic cheeses higher in preference over traditional cheese, albeit not significantly.     

Suitability of recombinant camel (Camelus dromedarius) chymosin as a coagulant for Cheddar cheese

Cheddar-type cheeses were manufactured using fermentation-produced camel or calf chymosin. There were no significant differences in the composition and pH between the cheeses made with either coagulant. The extent of primary proteolysis was significantly lower in cheeses made with camel chymosin than in cheeses made with calf chymosin. There were large quantitative differences between the peptide profiles of cheeses; however, the levels of amino acids were similar except for isoleucine, histidine and lysine. The cheeses made with camel chymosin were characterized by lower intensities of sulphur and brothy flavours and showed less bitter taste; however, the cheeses made with calf chymosin had greater breakdown of texture, higher smoothness and mouthcoating and were more cohesive and adhesive. The results of this study suggest that camel chymosin appears to be suitable for making Cheddar cheese with lower levels of proteolysis but with good flavour.     

Effect of heterofermentative lactic acid bacteria of DL-starters in initial ripening of semi-hard cheese

The role of heterofermentative lactic acid bacteria from dl-starters in ripening of semi-hard cheese was investigated using the strains Leuconostoc pseudomesenteroides PS12 and 1159, Leuconostoc mesenteroides subsp. cremoris T26 and Lactobacillus danicus 13M1. Control cheese was made with starter containing only homofermentative Lactococcus lactis subspecies. Leuc. mesenteroides subsp. cremoris T26 did not grow in cheese and started to decrease in number early, whereas the others grew and remained at a high number throughout the nine-week ripening period. None of the added heterofermentative strains affected proteolysis and total amount of amino acids; however, differences in the composition of amino acids were observed, and caused significant differences in the composition of volatile aroma compounds. Added strains increased the amount of secondary alcohols and mediated decreases in the amount of corresponding methyl ketones, diacetyl and acetoin. Eye formation was only affected by Lb. danicus through stimulation of late gas formation in cheeses.     

The effects of incorporating wild‐type strains of Lactococcus lactis into Turkish white‐brined cheese (Beyaz peynir) on the fatty acid and volatile content

The development of free fatty acids (FFA) and volatile flavour compounds in the Turkish white‐brined cheese Beyaz peynir made by using three wild strains of Lactococcus lactis subsp. lactis was investigated over 90 days. Results showed that production of both FFA and flavour compounds in the control (PK1) and experimental cheeses (MBLL9, MBLL23 and MBL27) was strain dependent. The hydrolysis of milk fat was more evident in the cheese made using Lc. lactis subsp. lactis MBL27. Considering the production of fat breakdown compounds and acidification activities of the strains MBLL23 and MBL27, the combination of these strains could be proposed for the production of white‐brined cheese.     

Effects of blends of camel and calf chymosin on proteolysis,residual coagulant activity,microstructure, and sensory characteristics of Beyaz peynir

Beyaz peynir, a white brined cheese, was manufactured using different blends of camel chymosin (100, 75, 50, 25, and 0%) with calf chymosin and ripened for 90 d. The purpose of this study was to determine the best mixture of coagulant for Beyaz peynir, in terms of proteolysis, texture, and melting characteristics. The cheeses were evaluated in terms of chemical composition, levels of proteolysis, total free amino acids, texture, meltability, residual coagulant activity, microstructure, and sensory properties during 90 d of ripening. Differences in the gross chemical composition were statistically significant for all types of cheeses. Levels of proteolysis were highly dependent on the blends of the coagulants. Higher proteolysis was observed in cheeses that used a higher ratio of calf chymosin. Differences in urea-PAGE and peptide profiles of each cheese were observed as well. Meltability values proportionally increased with the higher increasing levels of calf chymosin in the blend formula. These coagulants had a slight effect on the microstructure of cheeses. The cheese made with camel chymosin had a harder texture than calf chymosin cheese, and hardness values of all cheese samples decreased during ripening. The cheeses with a high ratio of calf chymosin had higher residual enzyme activity than those made with camel chymosin. No significant difference in sensory properties was observed among the cheeses. In conclusion, cheeses made with a high level of calf chymosin had a higher level of proteolysis, residual coagulant activity, and meltability. The cheeses also had a softer texture than cheeses made with a high content of camel chymosin. Camel chymosin may be used as a coagulant alone if low or limited levels of proteolysis are desired in cheese.     

Influence of a bacteriocin-producing lactic culture on proteolysis and texture of Hispánico cheese

Hispánico cheese was manufactured in duplicate experiments, each consisting of two 50-L vats, and ripened for 75 days. Lactic cultures for experimental cheese were 0.5% Lactococcus lactis subsp. lactis INIA 415 (Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, Madrid, Spain), a bacteriocin-producing (Bac⁺) strain harbouring the structural genes of nisin Z and lacticin 481, 0.5% L. lactis subsp. lactis INIA 415-2, a Bac⁻ mutant and 2% TA052, a commercial Streptococcus thermophilus culture. Lactic cultures for control cheese were 1% L. lactis subsp. lactis INIA 415-2 and 2% TA052. S. thermophilus counts were lower, and levels of cell-free intracellular aminopeptidases higher, from day 1 in cheese made with the bacteriocin producer, indicating early lysis of the thermophilic culture. Experimental cheese showed reduced proteolysis of α_s-casein and lower levels of hydrophilic and hydrophobic peptides than control cheese. However, proteolysis as estimated by the o-phthaldialdehyde method and total level of free amino acids were in experimental cheese 1.38- and 2.47-fold, respectively, those in control cheese on day 25, and 1.49- and 2.34-fold, respectively, on day 75. Higher values of fracturability, elasticity and hardness were recorded from day 50 for cheese made with the bacteriocin producer, which were related to its higher residual α_s-casein content. The use of a bacteriocin-producing culture, though retarding α_s-casein proteolysis and softening of texture, enhanced considerably secondary proteolysis during cheese ripening.     

Milk fermentation by functional mixed culture producing nisin Z and exopolysaccharides in a fresh cheese model

A nisin Z-producing strain, Lactococcus lactis subsp. lactis biovar. diacetylactis UL719 and two nisin-sensitive cultures, Lactobacillus rhamnosus RW-9595 M producing exopolysaccharide (EPS), and Lc. lactis subsp. cremoris for acidification, were tested in pure and mixed cultures during milk fermentation. The mixed culture of the three strains showed a higher acidifying capacity at 34°C and 38°C, even though populations of Lc. cremoris were largely reduced compared with pure cultures. Bacteriocin production was 3.1–4.6-fold higher in mixed cultures than for pure cultures of Lc. diacetylactis UL719. These data can be explained by commensalism behavior relying on high proteolytic activity of Lc. cremoris and autolysis and nisin Z-induced lysis. In mixed culture, EPS production was 3-fold lower than for Lb. rhamnosus RW-9595 M pure culture. Our data showed that this strain combination, with nisin-producing and sensitive strains, can be used in mixed cultures for manufacture of fresh cheese with improved functional properties.     

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.     

The effects of freezing and frozen storage of ewe's milk cheese on the viability and proteolytic activity of lactococci used as a starter

A study has been conducted on the effect of two freezing conditions (slow and fast) and frozen storage of ewe's milk cheese (?20 °C for 4 months) on the viability and the proteolytic (cascinolytic and aminopeptidase) activity ofLactococcus lactis subsp.lactis andL. lactis subsp.cremoris used as starters in the manufacture of cheese. The study was carried out on the lactococci subjected to freezing and frozen storage, either in the cheeses or in curds simulating a model system. As well as other parameters used to quantity microbial activity, the total viable counts during the subsequent cheese ripening have been analysed in the frozen cheeses. Frozen storage of the investigated lactococci, either in the cheese or in model systems, gave rise to significant decreases in the caseinolytic and aminopeptidase activities greater than did freezing alone. No clear differences were found in enzymatic activity values when using slow or fast freezing conditions. Storage of the cheeses under frozen conditions also affected microbial viability and consequently caused a greater decrease of the viable flora during subsequent ripening of the frozen stored cheeses.     

Dynamics of starter,adjunct non-starter lactic acid bacteria and propionic acid bacteria in low-fat and full-fat Dutch-type cheese

The microbial dynamics of Dutch-type cheeses differing in starter (commercial DL starter or single strain of Lactococcus lactis ssp. cremoris), adjunct (Lactobacillus or Propionibacterium) and fat contents (10% or 28% fat) were investigated by culture-dependent and culture-independent analysis. The cheese microbiota was dominated by the adjunct Lactobacillus after 4 weeks of ripening and the fat content did not influence the microbial diversity. The Leuconostoc sp., presumably from the DL starter, was detected in cheeses made with added Lactobacillus plantarum and Lactobacillus rhamnosus and was not detected in cheese made with added Lactobacillus paracasei after 4 and 7 weeks. No Lactobacillus spp. were detected in cheese with added Propionibacterium, while Leuconostoc was the only species detected. In cheeses made with Lc. lactis ssp. cremoris as starter, the Lactobacillus microbiota was similar to the cheese milk microbiota after 24 h while after 4 weeks different species of Lactobacillus and Leuconostoc were detected.     

Effects of variousMicrococcus strains on the ripening and organoleptic characteristics of Arzúa cow's-milk cheese

Ten batches of Arzúa, a soft cow's-milk cheese produced in northwest Spain, were prepared from pasteurized milk. Two (control) batches (CB) were made with a commercial starter containingLactococcus lactis subspecieslactis andcremoris. Another eight batches (MB) were made with the commercial starter plus one of eightMicrococcus spp. strains previously isolated from raw-milk Arzúa cheeses. In all MB, β-casein degradation over the 30-day ripening period was more pronounced (mean 31%) than in the CB (mean 12%). α_S1-Casein degradation was highly variable in the MB, though mean degradation over the ripening period (75%) was similar to that observed in the CB (73%). Similarly, volatile fatty acid (VFA) content was highly variable in the MB, with the mean content at 30 days (3.8 mEq per 100 g) being higher than in the CB (1.6 mEq per 100 g). Rheological characterization of the cheeses (by uniaxial compression) revealed significant differences between batches, with some samples fracturing under the compression pressure applied and others not. Sensory evaluation also revealed significant differences. “Non-milk” aromas were more frequently detected in batches made with lipolytic micrococcal strains. Betweenbatch differences in tastes and texture were also detected. Multiple correlation analysis of the data obtained at day 15 of ripening revealed statistically significant positive correlations (r>0.70) between α_S1-casein content and dry matter content, between α_S1-casein content and sensorially evaluated firmness, and between VFA content and sensorially evaluated rancidity. Statistically significant negative correlations between sensorially evaluated firmness and the ratio of α_S1-I content to α_S1-casein content were detected. The results of this study suggest a need for further studies aimed at selecting those strains which could be most suitable for the production of Arzüa cheeses; due to their effects on texture, α_S1-caseinolytic strains seem to be more appropriate than β-caseinolytic ones.     

Flavour enhancement of low-fat Feta-type cheese using a commercial adjunct culture

The effect of a commercial adjunct culture (CR-213), containing Lactococcus lactis subsp. cremoris and Lactococcus lactis subsp.lactis, added at the level of 0.06 or 0.09% (w/w) to cheese milk, on the characteristics of the resultant low-fat Feta-type cheese during aging, was studied. Two controls, a full-fat cheese (∼22% fat) and a low-fat cheese (∼7% fat, made using the standard procedure), were also prepared. The results indicated that the adjunct containing low-fat cheeses exhibited no significant (P>0.05) differences in compositional (moisture, fat, protein, salt, pH) or textural (force and compression to fracture, hardness) characteristics in comparison with the low-fat control cheese. It was also found that the use of the adjunct culture slightly improved the flavour intensity of the low-fat cheese which received a flavour score similar to that of the full-fat control cheese. Moreover, the experimental low-fat cheeses received significantly (P<0.05) higher total scores (overall quality) than the low-fat control cheese but lower than the full-fat cheese.     

Peptidase and proteinase activity ofLactococcus lactis,Lactobacillus casei andLactobacillus plantarum

The proteolytic system of several non-commercial strains of lactococci and lactobacilli that were isolated directly from traditional-Spanish, semi-hard, goats' milk cheese was studied. The aminopeptidase, X-prolyldipeptidyl aminopeptidase, dipeptidase and proteinase activity of these new strains was measured for the cytoplasmic, cell-wall/membrane and spontaneously released fractions. The aminopeptidase activity was exclusively intracellular and higher forLactobacillus casei subsp.casei than forLactococcus lactis subsp.lactis. Lactobacillus plantarum showed higher dipeptidase activity thanL. casei. The highest level of proteinase activity was recorded for the cell-wallmembrane fraction ofLactococcus lactis subsp.lactis IFPL 359, and was higher on β-casein than on α_s-casein for all the strains studied. These results suggest some different contribution of these strains to the proteolysis of cheese during ripening and they seem to complement each other when used together in the starter culture.     

Effects of high pressure on proteolytic enzymes in cheese: relationship with the proteolysis of ewe milk cheese

Ewe milk cheeses were submitted to 200, 300, 400, and 500 MPa (2P to 5P) at 2 stages of ripening (after 1 and 15 d of manufacturing; P1 and P15). The high-pressure-treated cheeses showed a more important hydrolysis of β-casein than control and 2P1 cheeses. Degradation of α_s1-casein was more important in 3P1, 4P1, and P15 cheeses than control and 2P1 cheeses. The 5P1 cheeses exhibited the lowest degradation of α_s-caseins, probably as a consequence of the inactivation of residual chymosin. Treatment at 300 MPa applied on the first day of ripening increased the peptidolytic activity, accelerating the secondary proteolysis of cheeses. The 3P1 cheeses had extensive peptide degradation and the highest content of free amino acids. Treatments at 500 MPa, however, decelerated the proteolysis of cheeses due to a reduction of microbial population and inactivation of enzymes.     

Use of Taiwanese ropy fermented milk (TRFM) and Lactococcus lactis subsp. cremoris isolated from TRFM in manufacturing of functional low-fat cheeses

Abstract: The purpose of this study was to manufacture new functional low‐fat cheeses using Taiwanese ropy fermented milk (TRFM) and Lactococcus lactis subsp. cremoris strains isolated from TRFM. After 28 d of ripening and storage, the viable populations of lactic acid bacteria (LAB) in the low‐fat cheeses made with L. lactis subsp. cremoris TL1 (TL1), L. lactis subsp. cremoris TL4 (TL4), and TRFM still maintained above 10⁸ CFU/g. The low‐fat cheeses made with TL1 and TRFM showed higher moisture contents than the cheeses made with TL4, full‐fat, and low‐fat cheese controls. The low‐fat cheeses made with TL1 and TL4 had higher customer preferential scores similar to full‐fat cheese control in the sensory evaluation. Additionally, the low‐fat cheeses fermented with TL1, TL4, and TRFM for 4 h had higher 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) free radical‐scavenging and ferrous ion‐chelating abilities than the cheeses fermented with the starters for 8 h, full‐fat, and low‐fat cheese controls. A better angiotensin‐converting enzyme (ACE) inhibition activity was also observed in the low‐fat cheeses made with TL1, TL4, and TRFM than that in the full‐fat and low‐fat cheese controls during ripening and storage period. Practical Application: As health‐conscious consumers continue to seek low‐fat alternatives in their diets, there remain strong interests for the dairy industry to develop low‐fat cheeses to meet the demands. This study clearly demonstrated that the low‐fat cheeses fermented with TL1 for 4 h showed a better overall acceptability and possessed antioxidative abilities and ACE inhibitory activities than other cheeses tested in this study. By improving its flavor and investigating the possible mechanisms of its functionalities in the future, this low‐fat cheese might possibly be commercialized and give a positive impact on cheese consumption in the future.     

Application of ruthenium red and colloidal gold-labeled lectin for the visualization of bacterial exopolysaccharides in Cheddar cheese matrix using transmission electron microscopy

The objective of the present study was to develop a methodology for direct observation of capsular and ropy strains and their exopolysaccharides (EPS) in a Cheddar cheese matrix. Cheddar cheeses with 50% reduced fat were made from milk containing 1.7% fat using mixed starter culture containing either capsule-forming Lactococcus lactis subsp. cremoris (SMQ-461) or ropy L. lactis subsp. cremoris (JRF-1) strains. Control cheese was made using the EPS-negative L. lactis subsp. cremoris (RBL132) strain. Following cheese pressing, samples were taken from each cheese treatment and examined by transmission electron microscopy (TEM). Samples were divided into two series: the first was prepared following the conventional methods (involving fixation, post fixation, dehydration and embedding in resin) and the second with added ruthenium red at 0.15% (w/v) during the fixation, post fixation and washing procedures. Gold-labeled lectin was also used for the visualization and localization of EPS in cheese matrix. Electron micrographs showed that ruthenium red makes it possible to visualize and enhance the resolution of the EPS in a Cheddar matrix compared with the conventional method. The EPS layer of the capsular strain appeared regular and evenly distributed around the cell, whereas the cell-associated EPS layer produced by the ropy strain was longer, more irregular (having a filamentous structure) and unevenly surrounded the cell. EPS released from the ropy strain appeared to form a network-like structure located principally in whey pockets and appeared to interact with the casein matrix and fat globule membrane. Labeling EPS by lectin conjugated to colloidal gold could only be performed with conventional preparation of cheese samples and appeared to react only with the cell surface rather than with liberated EPS. Besides their ability to bind water and increase cheese yield, capsular and ropy strains used in this study appear to have potential autolytic characteristics, which may have an impact on cheese proteolysis, texture and flavor quality.     

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.