Proteolysis In Cheese Research Articles

Analysis of the non-volatile components and volatile compounds of hydrolysates derived from unmatured cheese curd hydrolysis by different enzymes

Fifteen cheese protein hydrolysates were produced by using four different proteases. Then, the free amino acids (FAAs), molecular weight distribution (MWD), electronic tongue evaluation, and volatile compounds of the corresponding products were evaluated, respectively. The results suggested that 2SD had the strongest hydrolysis characteristic, followed by 6SD and FN. Samples hydrolyzed for less than 6 h or more than 18 h contained great defects of taste. Peptides with 150 Da–450 Da were mainly responsible for bitterness, saltiness, umami, and aftertaste in some enzyme hydrolysis. Under the same total enzyme concentration condition, the sample hydrolyzed by Flavourzyme and Neutrase for 18 h released more richness and less bitterness than the other systems, which were characterized by butter and cream odor. Notably, it was found for the first time that tetramethylpyrazine (TMP) was detected in cheese proteolysis with the highest content of 17.59 μg/g in Protease 2SD for 30 h. 2-Undecanone and acetoin played a key role in the flavor formation of the tested samples. Regarding the different chemical families of volatiles, acids were more abundant in the samples hydrolyzed by Protease 2SD and 6SD, while FN systems can achieve high ketone content.

Influence of different types of fermentation-produced chymosin on quality of soft cheeses

The disadvantage of soft cheeses is their short shelf life. In soft cheeses with a high moisture content, proteolysis occurs at a high rate, as a result of which the cheeses quickly overripe. The main proteolytic agent in soft cheeses is the milk-clotting enzyme (MCE). Increasing the shelf life of cheeses can be achieved by using MCE types having low proteolytic activity (PA). We have studied the effect of MCE based on different types of fermentation-produced chymosin: Chy-max® Extra (bovine chymosin), Chy-max® M (camel chymosin), Chy-max® Supreme (“modified” chymosin) on the dynamics of proteolysis in soft cheeses and related changes in the structure of cheeses during their storage. All 3 types of studied MCEs have different levels of nonspecific PA. The higher the level of nonspecific PA of the used MCE, the higher the rate of the proteolysis process in the resulting cheeses. Increasing the dose of MCE also increases the rate of proteolysis in cheeses. To increase the shelf life of soft cheeses, which depends on the period of preservation of a dense consistency, it is promising to use MCE with low PA based on camel chymosin (Chy-max® M) and “modified” chymosin (Chy-max® Supreme).

Effects of Monascus on Proteolysis, Lipolysis, and Volatile Compounds of Camembert-Type Cheese during Ripening.

In order to improve the flavor and taste of Camembert cheese, the use of Monascus as an adjunct starter for the production of Camembert-type cheese was studied to investigate its effect on the proteolysis, lipolysis, and volatile compounds during ripening for 40 days. The Camembert cheese without Monascus was used as a control. The results showed that proteolytic and lipolytic activities increased to a certain extent. The addition of Monascus promoted primary and secondary proteolysis, due to the release of some proteases by Monascus. Aspartic, Threonine, Glutamic, Glycine, Methione, Isoleucine, Phenyalanine, and Lysine contents in experimental group (R) cheese were significantly higher than those in control group (W) cheeses. In addition, the free amino acid and fatty acid contents were also affected. The identification of flavor components using gas-mass spectrometry (GC-MS) showed that 2-undecone, 2-tridecanone, phenylethyl alcohol, butanediol (responsible for the production of flowery and honey-like aroma), ethyl hexanoate, ethyl octanoate, and ethyl citrate (fruit-like aroma) were significantly higher (p < 0.05) in the experimental cheeses than in the control. The contents of 2-nonanone, 2-octanone and 2-decanone (showing milky flavor), and 1-octene-3 alcohol with typical mushroom-like flavor were lower than the control.

Supplementing goats' diet with sainfoin pellets (versus alfalfa) modifies cheese sensory properties and fatty acid profile

In two different goat farms, producing Picodon and Mutschli cheese respectively, two groups of lactating goats were fed alfalfa pellets (ALF) or sainfoin pellets (SNF) for 6 weeks. The cheeses produced at the end of the trial were analysed for sensory properties, composition, fatty acid profile, proteolysis, and rheology. The SNF Picodon had higher “goat” and “farm” aroma and odour, and higher bitter taste, compared with the ALF Picodon. The SNF Picodon showed lower dry matter than the ALF Picodon, leading to differences in texture and mouthfeel. The SNF Mutschli had a different mouthfeel than the ALF Mutschli. In a triangle test, consumers were able to perceive a difference in both Picodon and Mutschli between ALF and SNF groups. In both Picodon and Mutschli, feeding sainfoin increased cheese proteolysis and modified cheese fatty acid profile, resulting in higher polyunsaturated fatty acids proportion and lower n-6/n-3 ratio.

Characteristics and proteolysis of a Spanish blue cheese made with raw or pasteurised milk

Valdeón cheese is a Spanish Protected Geographical Indication of blue‐veined cheese produced on an industrial scale from raw or pasteurised cow and/or goat milk. The aim of this work was to evaluate the impact of pasteurisation on the microbiological, physicochemical and sensory characteristics of cheese. Cheeses made with raw milk showed higher counts of lactobacilli and enterococci, as well as lower values of pH and D‐lactic acid and salt/moisture ratio. Cheeses made with pasteurised milk showed greater extent of proteolysis, higher concentration of free amino acids and lower concentration of biogenic amines. Pasteurisation produced more elastic and harder cheeses, but with a lower sensory profile.

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

Cheddar-type cheese was manufactured using fermentation-produced bovine chymosin (BC), fermentation-produced camel chymosin (CC) and a modified fermentation-produced camel chymosin (mCC) and ripened for 180 days. Only minor differences were found in cheese composition and pH between the cheeses made with any of the chymosins studied. Proteolysis in cheese made with mCC was reduced compared with cheese made with BC or CC. Significantly higher instrumental and sensory hardness and significantly lower meltability were found in cheeses made using CC or mCC compared with cheese BC after 180 days of ripening. Descriptive sensory analysis results showed that cheese made with CC or mCC had less sulphur and barny flavour; the brothy flavour and bitter taste of cheese made with mCC were also lowest. In conclusion, the modified camel chymosin appears to be suitable for the manufacture of Cheddar cheese with modified functionalities.

Effects of probiotic adjuncts on physicochemical properties, organic acids content, and proteolysis in cheese prepared from buffalo milk

The aim of the current study was to assess the effects of probiotic adjuncts on physiochemical characteristics, organic acids, water-soluble nitrogen percentage (WSN%), total free amino acids (TFAA), and proteolytic patterns in buffalo milk probiotic Cheddar cheese during 120 days of ripening at 4°C. Cheddar cheese was prepared with commercially available mesophilic cheese starter cultures (control), Lactobacillus acidophilus, and Bifidobacterium bifidum individually as well as in synergistic manner to prepare probiotic cheese. The results appeared with a significant difference in control and probiotic cheese with reference to citric, lactic, butyric, and acetic acids, WSN%, and TFAA production. The proteolytic pattern of probiotic bacteria modified the proteolysis and expedited the formation of bioactive peptides in Cheddar cheese. Among probiotic adjuncts, Cheddar having B. bifidum exhibited higher WSN% and TFAA as compared to the other two probiotic cheese samples. Hence, it can be concluded that B. bifidum is slightly more proteolytic as compared to other probiotic adjuncts and the incorporation of probiotic adjuncts enhanced the hydrolysis of αs1− casein in Cheddar cheese. Practical applications Bioactive peptides remain inactive within the protein molecule. When milk undergoes enzymatic hydrolysis (protein), fermentation (through the proteolytic starter), or enzymatic proteolysis, bioactive compounds become active and capable to exhibit their therapeutic potential. Probiotic adjuncts modify the proteolysis, balance bio-diversity removed by pasteurization and expedite the formation of bioactive peptides in Cheddar cheese. Regarding probiotic adjuncts, cheese serves as one of the most efficient carriers of probiotic bacteria owing to its matrix efficiency and ability to maintain the viability of bacteria and facilitate their entry into the human gut.

Characterization of autochthonal yeasts isolated from Spanish soft raw ewe milk protected designation of origin cheeses for technological application

The yeasts involved in the ripening process of artisanal soft raw ewe milk Protected Designation of Origin (PDO) Torta del Casar and Queso de la Serena cheeses produced in Extremadura, Spain, were isolated throughout their ripening process, strain typed, and characterized for some important technological properties. A total of 508 yeast isolates were obtained and identified by inter-single sequence repeat anchored PCR amplification analysis and subsequent sequencing of the internal transcribed spacer ITS1/ITS2 5.8S rRNA. A total of 19 yeast species representing 8 genera were identified. Debaryomyces hansenii, Pichia kudriavzevii, Kluyveromyces lactis, and Yarrowia lipolytica were the predominant species. We selected 157 isolates, by genotyping and origin, for technological characterization. The evaluation of yeast isolates' growth under stress conditions of cheese ripening showed that 87 presented better performance. Among them, 71 isolates were not able to catabolize tyrosine to produce a brown pigment. Principal component analysis of the biochemical features of these isolates showed that 9 strains stood out, 3 K. lactis strains (2287, 2725, and 1507), 2 Pichia jadinii (1731 and 433), 2 Yarrowia alimentaria (1204 and 2150), Y. lipolytica 2495 and P. kudriavzevii 373. These strains displayed strong extracellular proteolytic activity on skim milk agar as well as an adequate enzymatic profile (strong aminopeptidase and weak protease activity), suggesting their great potential for cheese proteolysis. Extracellular lipolytic activity was mainly restricted to Yarrowia spp. isolates and weakly present in P. kudriavzevii 373 and K. lactis 2725, although enzymatic characterization by API-ZYM (bioMérieux SA) evidenced that all may contribute, at least in part, to the lipolysis process. Moreover, these strains were able to assimilate lactose, galactose, and glucose at NaCl concentrations higher than that usually found in cheese. However, lactate and citrate assimilation were limited to Y. lipolytica 2495, P. kudriavzevii 373, and P. jadinii 433, and may contribute to the alkalinizing process relevant to biochemical processes that take place in the last stages of ripening. By contrast, K. lactis strains showed acidifying capacity and β-galactosidase activity and may take part in the initial stages of ripening, together with lactic acid bacteria. Thus, considering the technological characteristics studied, the 9 selected strains presented biochemical features well suited to their potential use as adjunct cultures, alone or in combination with autochthonous starter bacteria in the cheesemaking process, to overcome the heterogeneity of these PDO cheeses, preserving their unique sensory characteristics.

Low-fat cheese in the focus of dietary nutrition

Currently, products with a reduced content of saturated fatty acids and a high protein content occupy an important place in the structure of a balanced diet, which leads to an increasing demand for cheeses with a low content of milk fat and high organoleptic properties. As a result of the growing trend towards the consumption of products with a reduced calorie content, there is a scientific interest in the development of technologies for low-fat cheeses with high consumer characteristics that are not inferior to analogues with a fat content of 45-50%. However, low-fat cheeses produced using existing technologies, as a rule, are characterized by low organoleptic properties, including an unexpressed taste and aroma and a rough, rubbery texture. The purpose of the research was to develop a low-fat cheese with high organoleptic characteristics, produced using not only the main acid-forming microflora, but also adjunct bacterial cultures that enhance enzymatic processes during cheese ripening and have probiotic properties. Material and methods. The production of low-fat cheeses (options 2 and 3) with a fat mass fraction of 20% in terms of dry matter was carried out according to a single technological scheme, which ensures the manufacture of cheese after pressing with a mass fraction of moisture of 53-54%. As a control option (1), Dutch cheese was produced with a fat mass fraction of 45%. Bulk starter containing a mixture of lactococci of the Lactococcus lactis species was used as the main acid-forming starter microflora in all cheese options (1, 2, 3); in low-fat cheese option 3, adjunct cultures of Lactobacillus casei and Propionibacterium freudenreichii were added. During ripening, cheeses were subjected to microbiological (total quantity of viable cells of lactic acid microorganisms, Lactobacillus casei and Propionibacterium freudenreichii), physicochemical (mass fraction of lactose, fat, moisture, solids, protein) and organoleptic studies. The total quantity of viable cells of the mesophilic lactic acid microflora was determined by the quantity of mesophilic aerobic and facultative anaerobic microorganisms. In cheeses after the end of the ripening process, the molecular mass distribution of soluble nitrogenous compounds and the content of flavoring substances in the vapor phase were additionally evaluated. Results. It has been established that in low-fat cheeses, the mass fraction of protein increases by 5.9±0.1% with a decrease in the proportion of fat to 20.0%. In this regard, the calorie content of low-fat cheese reduced by a third. It has been demonstrated that differences in the composition of the starter microflora had a significant impact on the depth and direction of biochemical processes, the formation of flavoring substances, which lead to an improvement in the organoleptic characteristics of cheeses. Conclusion. The addition of adjunct cultures of Lactobacillus casei mesophilic rods and Propionibacterium freudenreichii propionic acid bacteria, together with technological methods, contributes to the formation of a pronounced cheese taste and aroma, improves the flavor profile and deepens the process of proteolysis in cheeses with a fat mass fraction of 20% and brings their consumer properties closer to those of cheese with a fat content of 45%.

The effects of small changes in temperature on proteolysis during isothermal and non-isothermal aging of full-fat and reduced-fat Cheddar cheese

&lt;abstract&gt; &lt;p&gt;Effective approaches for modulating the evolution of cheese quality attributes are needed for mitigating challenges that are associated with fluctuating supply and demand as well as with disrupt supply chain. Proteolysis is the most important and most complex cascade of events that affects the evolution of cheese quality attributes. Information about the effects of small changes in temperature during isothermal and non-isothermal aging of Cheddar cheese at temperatures lower than 10 ℃ on proteolysis has been developed to a very limited extent. The objective of the research was to age FF and RF Cheddar cheeses for six months at different isothermal and non-isothermal time-at-temperature regimes at temperature ranging from 5 to 8 ℃ and to investigate the effects of these conditions on proteolysis. Changes in the level of cheese-N fractions that are soluble at pH 4.6, soluble in 12% TCA and soluble in 5% PTA were monitored. The proteolytic cascade during aging was significantly (p &amp;lt; 0.05) influenced by a combined impact of the time-at-temperature details of aging and cheese composition. The highest and lowest levels of the investigated fractions were found in cheeses that had been aged isothermally at 8 and 5 ℃, respectively. In most cases, proteolysis in the FF cheeses was to a higher extent than in the RF ones. Proteolysis during non-isothermal aging was significantly affected by the aging regime in a time-at-temperature-specific manner (p &amp;lt; 0.05). The results can offer new opportunities for modulating the rate of cheese aging. The demonstrated significant effect of a very small change in aging temperature on proteolysis during cheese aging also highlights the critical importance of establishing and maintaining isotropic temperature distribution in cheese aging rooms.&lt;/p&gt; &lt;/abstract&gt;

Evaluation of Proteolysis, Antioxidant and Ace-Inhibitory Activities in a Model Cheese: Effect of Adjunct Lactobacilli, Ripening Time and Temperature

Evaluation of Proteolysis, Antioxidant and Ace-Inhibitory Activities in a Model Cheese: Effect of Adjunct Lactobacilli, Ripening Time and Temperature

Effect of heat treatment on whey protein-reduced micellar casein concentrate: A study of texture, proteolysis levels and volatile profiles of Cheddar cheeses produced therefrom

Micellar casein concentrate (MCC) of high casein content (93.64% of total protein), was produced by microfiltration of pasteurised skim milk. Cheesemilk of typical (1 × ) or high (1.5 × ) casein content were formulated from MCC which received either: no further heat treatment; pasteurisation (72 °C × 15 s) or high heat treatment (90 °C × 15 s), prior to combination with other membrane streams and cream. Cheddar cheeses were manufactured in triplicate and ripened for 180 days. Cheeses made from pasteurised or high heat treated MCC had similar pH as well as comparable flowability, hardness and volatile profiles over 180 days of ripening. On increasing cheesemilk casein content, levels of primary proteolysis in resultant cheeses decreased, and pH and hardness levels increased. Overall, increasing the heat treatment temperature from 72 to 90 °C for extension of the microbial shelf life of MCC does not impair the texture, functionality and volatile profile of resultant Cheddar cheeses.

Ripening Changes of the Chemical Composition, Proteolysis, and Lipolysis of a Hair Sheep Milk Mexican Manchego-Style Cheese: Effect of Nano-Emulsified Curcumin.

The influence of nano-emulsified curcumin (NEC) added to the hair sheep milk, prior to cheese-making, on the chemical composition, lipolysis, and proteolysis of manchego-style cheeses were evaluated throughout 80 days of ripening. The addition of NEC to the milk resulted in cheeses with the same moisture content (42.23%), total protein (23.16%), and water activity (0.969) (p > 0.05). However, it increased the fat and ash levels from 26.82% and 3.64% in B 10 ppm to 30.08% and 3.85% in C 10 ppm, respectively, at the end of the ripening (p < 0.05). The total phenolic content and antioxidant activity of experimental cheeses increased during ripening, and the fatty acid groups showed significant changes occurred to a greater extent in the first days of ripening (p < 0.05). The lipolysis increased consistently in all cheeses until day 40 of ripening, to decrease at the end, while proteolysis increased during all ripening time in all samples (p < 0.05); the addition of NEC did not alter the primary proteolysis of manchego-style cheeses, but it modified secondary proteolysis and lipolysis (p < 0.05). Principal component analysis was useful for discriminating cheeses according to their chemical composition and classified into four groups according to their ripening time. This research highlights the potential of CNE to fortify dairy foods to enhance their functionality.

The effects of camel chymosin and Withania coagulans extract on camel and bovine milk cheeses

Withania coagulans (W. coagulans) extract and camel chymosin have aspartic protease capable of coagulating milk for cheese production. This study investigated the quality of camel and bovine milk cheeses coagulated using Withania extracts, came chymosin, and their mixture in two experiments. In Experiment (1), a factorial design with four factors (W. coagulans, camel chymosin, incubation time, and incubation temperature) was performed. The effect of these factors on cheese’s yield and hardness were assessed. An enzyme concentration corresponding to a 36 µg/L of milk of W. coagulans, 50 IMCU/L of camel chymosin, holding time of 4 h, and incubation temperature of 60 °C provided the optimal textural hardness for both camel and bovine milk cheeses. Seven treatments were analyzed in experiment (2) were analyzed for physicochemical properties, yield, and sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGEitation). The results showed that pure Withania extract exhibited the lower coagulating effect resulting in cheeses with low yield, hardness, fat, protein, and total solids. The SDS-PAGE electropherograms of camel cheese showed several low molecular weight bands as compared to bovine cheese. This phenomenon is due to excessive proteolysis in camel cheese, which we believed is caused by the presence of endogenous enzymes.

Spectrophotometric method for evaluating proteolysis in cheeses and aromatic additives with a cheesy taste

The spectrophotometric method for measuring protein content can be used to evaluate the degree of proteolysis in cheeses. At a wavelength of 280 nm, tryptophan and tyrosine are absorbed, a high amount of them is found in casein, the main protein of cheese mass. It was found that the value of the absorbance coefficient of the solution of proteins extracted from flavoring additives with cheese flavor (FA) and cheeses depends on the degree of proteolysis of proteins in the cheese mass and differs in FA and different types of cheeses. The highest absorbance coefficient is observed in the FA samples A1%1cm = 10.30, in which from 65 to 81% of the protein is converted into a soluble state. In cheeses, the degree of proteolysis is from 23 to 33%, and the absorbance coefficient of solution is from 1.1 to 2.4 (with the exception of Cheddar cheese), which indicates an incomplete transition of amino acids absorbing radiation at 280 nm into the extract released from cheeses. Using the spectrophotometric method, the results of measuring the content of soluble protein in cheeses and FA, strictly correlating with the results achieved by the Kjeldahl method (R2 &gt; 0.81), can be obtained. To get reliable results of evaluating the content of water-soluble protein in cheeses, it is necessary to carry out measurements on a sample of cheeses belonging to the same species group, having the same specificity of proteolysis and slightly different absorbance coefficient between samples within the instance.

Optimizing the acceleration of Cheddar cheese ripening using response surface methodology by microbial protease without altering its quality features

Cheddar cheese proteolysis were accelerated employing Penicillium candidum PCA1/TT031 protease into cheese curd. In the present study, several of the significant factors such as protease purification factor (PF), protease concentration and ripening time were optimized via the response surface methodology (RSM). The ideal accelerated Cheddar cheese environment consisted of 3.12 PF, 0.01% (v/v) protease concentration and 0.6/3 months ripening time at 10 °C. The RSM models was verified to be the most proper methodology for the maintain of chosen Cheddar cheese. Under this experimental environment, the pH, acid degree value (ADV), moisture, water activity (aw), soluble nitrogen (SN)%, fat and overall acceptability were found to be 5.4, 6.6, 35%, 0.9348, 18.8%, 34% and 13.6, respectively of ideal Cheddar cheese. Furthermore, the predicted and experimental results were in significant agreement, which confirmed the validity and reliability of the suggested method. In spite of the difference between the ideal and commercial Cheddar cheese in the concentration of some of amino acids and free fatty acids, the sensory evaluation did not show any significant difference in aroma profile between them.

Proteolysis and ACE-inhibitory peptide profile of Cheddar cheese: Effect of digestion treatment and different probiotics

Proteolysis degree, ACE-inhibitory (ACE-I) activity, water-soluble peptide profiles of Cheddar cheese with different probiotics were determined during digestion period and were comprehensively compared to evaluate the influence of digestion and the addition of different probiotics. After digestion, further hydrolysis of protein led to the production of more low molecular weight peptides, and ACE-I peptides with higher activity were released. ACE-I activity of Cheddar cheese with Lactobacillus helveticus (B4, 86.06%) was significantly (P < 0.05) higher than other cheese groups. Three ACE-I peptides were uniquely presented in B4 before digestion and more new ACE-I peptides were released after digestion. Meanwhile, cheese with Lactobacillus rhamnosus was more likely to produce anti-digestion ACE-I peptides. Most of ACE-I peptides come from β-casein. Hydrophobic amino acids Val, Phe, Pro, Tyr, Leu, and basic amino acids Arg, Lys at the C-terminal were more responsible for the formation of ACE inhibitory peptides that resist digestion. This research provides the basis to increase the exploitation of the health benefits of Cheddar cheese with probiotic.

Effect of probiotic Minas Frescal cheese on the volatile compound and metabolic profiles assessed by nuclear magnetic resonance spectroscopy and chemometric tools

This study aimed to evaluate the effect of Lacticaseibacillus casei 01 as a probiotic culture on the production of volatile organic compounds and metabolic profile of Minas Frescal cheese. Lactose (α-lactose and β-lactose), fatty acids (unsaturated and saturated), citric acid, tryptophan, and benzoic acid were the main compounds. Compared with the control cheese, probiotic cheese was characterized by the highest concentration of tryptophan and presented a higher number of volatile acids. The control cheese was characterized by the highest concentration of benzoic acid and fatty acids, resulting in a higher number of volatile alcohols and esters. No differences were observed for α-lactose, β-lactose, and citric acid contents. A clear separation of probiotic and control Minas Frescal cheese was obtained using 1H nuclear magnetic resonance spectra, demonstrating that the addition of probiotic culture altered the metabolic profile of Minas Frescal cheese. Overall, the findings suggested that the addition of probiotic culture promoted the proteolysis in the fresh cheeses, decreased the lipolysis, and altered the volatile compounds. Furthermore, nuclear magnetic resonance spectroscopy coupled to chemometrics tools could be used to differentiate probiotic and conventional cheeses.

Validation of a reversed‐phase high‐performance liquid chromatographic method for the quantification of primary proteolysis during cheese maturation

A new fast and reliable reversed‐phase high‐performance liquid chromatographic method is proposed for quantifying all the major casein fractions during cheese proteolysis as an alternative to more complex methodologies. Excellent separation of the caseins, para‐κ‐casein (ρ‐κ‐CN), αs1‐casein (αs1‐CN), αs1‐I‐casein (αs1‐I‐CN), αs2‐casein (αs2‐CN), and β‐casein (β‐CN) was achieved, in terms of linearity, reproducibility, repeatability, accuracy and certified standard recovery. Validation by quantification of semihard and Maasdam‐style cheeses gave comparable results for ρ‐κ‐CN, β‐CN and αs‐casein (αs‐CN) to published data for both urea–polyacrylamide gel electrophoresis and capillary electrophoresis.

Effects of shredding on the functionality, microstructure and proteolysis of low-moisture mozzarella cheese

Low-moisture mozzarella cheese (LMMC) is commonly shredded before packaging, however, the effects of shredding are not fully understood. Industrially-produced block and shredded LMMC were studied during 8 weeks of storage at 4 °C. Cheese shredded on 15 d and at 8 weeks of age, coated with microcrystalline cellulose and stored in a modified atmosphere (70% N2 and 30% CO2), had an altered microstructure after 8 weeks compared with vacuum-packed block cheese. In the latter case the fat formed a more dispersed phase. Proteolysis was higher in shredded samples and a higher level of two bacterial proteases was detected. Despite these differences, the meltability and stretchability of the block and shredded LMMC were similar. The microstructure and functionality of cheese shredded at 15 d and stored for a further 6 weeks was similar to cheese shredded at 8 weeks, suggesting there is a flexible period for performing cheese shredding processes.