Lipolysis,proteolysis and sensory properties of ewe’s raw milk cheese (Idiazabal) made with lipase addition

In this paper, we describe the effect of the addition of pregastric lipase on the composition and sensory properties of Idiazabal cheese. Free fatty acids (FFA), partial glycerides, free amino acids (FAA), gross composition and sensory characteristics were determined at different ripening times in cheeses manufactured with three different amounts of commercial animal lipase or with lipase-containing artisanal lamb rennet paste. The addition of lipase increased the content of total FFA, particularly of short-chain FFA, and that of total partial glycerides in cheeses. Unexpectedly, lipase utilization significantly affected total FAA concentration, which decreased in cheeses elaborated with high lipase amount. In general, Val, Glu and Leu were the major FAA, and their concentrations depended, mainly, on ripening time. Lipase addition had significant influence on the sensory characteristics of the cheeses, increasing scores for most of the flavour and odour attributes of the cheese. Principal component analysis (PCA) was done including dry matter, FFA, FAA, partial glycerides and odour and flavour attributes of the cheeses. It indicated that aroma and flavour parameters of Idiazabal cheese and the content of short-chain FFA and diglycerides were highly correlated to first principal component (PC1), while texture parameters, compositional variables and FAA were correlated to the second principal component (PC2).     

Ripening of Camembert-type cheese made from caprine milk using calf rennet or kid rennet as coagulant

Camembert-type cheese was made from caprine milk using either calf rennet or kid 'Grandine' rennet as coagulant. The pH of all cheeses increased throughout ripening and levels of pH 4.6-soluble nitrogen increased from 8.1 to 18.2% of total nitrogen (TN) and from 6.9 to 20% TN for the cheeses made using calf rennet and kid rennet, respectively. Degradation of β-casein, measured by urea–polyacrylamide gel electrophoresis, and total and free amino acids were greater in the cheese made using kid rennet. Production of peptides, analysed by high performance liquid chromatography (HPLC), was slightly more extensive in the Camembert-type cheese made using calf rennet as coagulant. In general, a higher degree of proteolysis was found in Camembert-type cheese made from caprine milk using kid rennet than in cheese made using calf rennet as coagulant.     

Proteolysis in reduced sodium Feta cheese made by partial substitution of NaCl by KCl

Feta cheeses (five trials) of different sodium content were made, using ewes’ milk, from split lots of curd by varying the salting procedure, i.e. dry salting with NaCl (control) or mixtures of NaCl/KCl (3:1 or 1:1, w/w basis) and filling the cans with brine made with NaCl or the above NaCl/KCl mixtures, respectively, in order to study the influence of the partial substitution of NaCl by KCl on the proteolysis during cheese ripening. The extent and characteristics of proteolysis in the cheeses were monitored during aging by using Kjeldahl determination of soluble nitrogen fractions (water-soluble nitrogen, trichloroacetic acid-soluble nitrogen, phosphotungstic acid-soluble nitrogen), the cadmium–ninhydrin method for the determination of total free amino acids (FAA), urea–polyacrylamide gel electrophoresis of cheese proteins followed by densitometric analysis of the α_s1- and β-casein fractions, reverse-phase HPLC analysis of the water-soluble extracts of cheeses, and ion-exchange HPLC analysis of FAA. The results showed that proteolysis was similar in control and experimental cheeses at all sampling ages, indicating that the partial substitution of NaCl by KCl in the manufacture of Feta cheese had no significant effect on the extent and characteristics of proteolysis during cheese aging.     

Changes in the free amino acid content during ripening of Idiazabal cheese: influence of starter and rennet type

The effect of starter and rennet type on free amino acid release during ripening of Idiazabal cheese was studied. Four batches of cheeses were manufactured depending on the rennet used, commercial calf rennet or artisanal lamb rennet, and the addition or not of starter culture. Cheese samples contained 24 individual free amino acids Leu, Glu, Val and Phe showing the highest contents during the ripening. The results indicated that the release of the free amino acids during ripening was strongly affected by starter added to the cheeses, and that this effect varied markedly with the rennet used for cheesemaking. Total amounts of free amino acids were higher for the cheeses made with commercial calf rennet than for those made with artisanal lamb rennet, regardless of starter addition. Likewise, the highest total free amino acid levels were found in the cheeses made with starter, regardless of the type of rennet used.     

Free amino acid content of goat's milk cheese made with animal rennet and plant coagulant

BACKGROUND: Enzymes present in the flowers of Cynara cardunculus (cyprosins) are used in the production of some traditional Spanish and Portuguese cheeses, replacing animal rennet. The aim of this work was to study the changes that take place in free amino acids during the ripening of a goat's milk cheese (Murcia al Vino) manufactured with plant coagulant (PC) or animal rennet (AR). RESULTS: The total free amino acid (TFAA) concentration increased during ripening, with Ile, Val, Ala, Phe, Gaba, Arg and Lys representing more than 50% of the TFAA content at 60 days in both types of cheese. The TFAA concentration was significantly higher in cheeses made with PC (854 mg 100 g^?1 total solids (TS)) than those made with AR (735 mg 100 g^?1 TS). The concentration of most free amino acids, especially His, Ser, Gln, Thr, Ala, Met and Ile, was higher in the PC cheese. CONCLUSION: Cheese made using PC as coagulant presented higher contents of free amino acid throughout the ripening period than cheese made using AR. Therefore we can conclude that the use of PC to produce Murcia al Vino goat's cheese would accelerate the ripening process as a result of increased cyprosin proteolytic activity. Copyright © 2011 Society of Chemical Industry     

Proteolysis in reduced sodium Kefalograviera cheese made by partial replacement of NaCl with KCl

Kefalograviera cheeses (five trials) of different sodium content were made from split lots of curd by varying the salting processes, i.e. brine — and dry — salting, with NaCl (control) or a mixture of NaCl/KCl (3:1 or 1:1, w/w basis). The extent and characteristics of proteolysis in the cheeses were monitored during aging by Kjeldahl determination of soluble nitrogen fractions (water-soluble nitrogen [WSN], trichloroacetic acid [TCA]-SN, phosphotungstic acid [PTA]-SN), the cadmium-ninhydrin method for the determination of total free amino acids (FAA), urea-polyacrylamide gel electrophoresis of cheese proteins, followed by densitometric analysis of the α_s1- and β-casein fractions, reverse-phase high-performance liquid chromatography (HPLC) analysis of the water-soluble extracts of cheeses, and ion-exchange HPLC analysis of FAA. The results showed that proteolysis was similar in control and experimental cheeses at all sampling ages, indicating that the partial substitution of NaCl by KCl in the manufacture of Kefalograviera cheese did not significantly influence the extent and characteristics of proteolysis during cheese aging.     

Effect of partial substitution of caprine milk for ovine milk,dry salting and cured scalding on lipolysis in Urfa cheeses

The objectives of this study were to determine the effect of partial substitution of caprine for ovine milk, dry salting and curd scalding on the free fatty acid (FFA) level of Urfa cheeses. At the end of storage, lauric and linoleic acids were lower in cheese made from milk where up to the 30% partial substitution of caprine milk for ovine milk was made. Dry salted cheeses had higher butyric, lauric, linoleic and linolenic acid levels, and lower palmitic acid levels than their scalded counterpart. A relative increase in short‐chain FFAs occurred during ripening. Palmitic and oleic acids were the most abundant FFAs in fresh and ripened Urfa cheeses.     

Evolution of free amino acids during ripening of Caciocavallo cheeses made with different milks

The evolution of free amino acids (FAA) in Caciocavallo cheeses, made with cow milk (CC) and cow milk mixed with ewe (CE) and goat (CG) milk, was studied throughout ripening. In all Caciocavallo cheeses produced, the total free amino acid (TFAA) content increased during ripening. In general, the highest TFAA content was found in cow cheeses, and the lowest in CG cheeses, whereas CE cheeses ranged over an intermediate level. In all the analyzed samples, during ripening, the content of the individual FAA increased with the exception of arginine. Tyrosine and histidine were found only in CE samples from the middle to the end of ripening. The major FAA found throughout the whole ripening period, in all types of cheese, were leucine, phenylalanine, lysine, valine, asparagine, γ-aminobutyric acid, and ornithine. The TFAA and several AA showed significant differences in ripening time, whereas tyrosine and histidine showed significant differences in kinds of milk.     

Probiotic in lamb rennet paste enhances rennet lipolytic activity, and conjugated linoleic acid and linoleic acid content in Pecorino cheese

Cheeses manufactured using traditional lamb rennet paste, lamb rennet paste containing Lactobacillus acidophilus, and lamb rennet paste containing a mix of Bifidobacterium lactis and Bifidobacterium longum were characterized for the lipolytic pattern during ripening. Lipase activity of lamb rennet paste, lamb rennet containing Lb. acidophilus, and lamb rennet containing a mix of bifidobacteria was measured in sheep milk cream substrate. Rennet paste containing probiotics showed a lipase activity 2-fold greater than that displayed by traditional rennet. Total free fatty acid (FFA) in sheep milk cream was lower in lamb rennet paste (981 μg/g of milk cream) than in lamb rennet containing Lb. acidophilus (1,382.4 μg/g of milk cream) and in lamb rennet containing a mix of bifidobacteria (1,227.5 μg/g of milk cream) according to lipase activity of lamb rennet paste. The major increase of FFA in all cheeses occurred during the first 30 d of ripening with the greatest values being observed for C16:0, C18:0 C18:1. At 60 d of ripening all cheeses showed a reduction in the amount of free fatty acids; in particular, total free fatty acids underwent a decrease of more than 30% from 30 to 60 d in cheeses manufactured using traditional lamb rennet paste, whereas the same parameter decreased 10% in cheeses manufactured using lamb rennet paste containing Lb. acidophilus and cheeses manufactured using lamb rennet paste containing a mix of B. lactis and B. longum. Cheese containing Lb. acidophilus was characterized by the greatest levels of total conjugated linoleic acids (CLA) 9-cis, 11-trans CLA and 9-trans, 11-trans CLA, whereas cheese containing bifidobacteria displayed the greatest levels of free linoleic acid. Rennet pastes containing viable cells of Lb. acidophilus and a mix of B. lactis and B. longum were able to influence the amount of FFA and CLA in Pecorino cheese during ripening.     

Texture and flavour development in Ras cheese made from raw and pasteurised milk

Texture, proteolysis and flavour development in Ras cheeses made from raw or pasteurised milk with two different thermophilic lactic cultures were monitored during ripening. Results showed that at day 1 of manufacture, the moisture content and pH were lower in raw milk cheese than in pasteurised milk cheeses. Levels of water-soluble nitrogen, casein breakdown, free amino groups and free fatty acids were higher in cheese made from raw milk than in that made from pasteurised milk. Textural characteristics, such as hardness, cohesiveness and chewines, increased in all treatments during the first 60 days of ripening due to the reduction in the moisture level during the second stage of salting (dry salting during the first 60 days of ripening). Cheese made from raw milk received the highest texture and flavour scores by panellists.     

Influence of lamb rennet paste on composition and proteolysis during ripening of Pecorino foggiano cheese

Rennet pastes produced by lambs subjected to three different feeding systems (mother suckling [MS], artificial rearing [AR], and artificial rearing with Lactobacillus acidophilus supplementation [ARLB]) and slaughtered at two different ages (20 and 40 d) were used for the manufacture of Pecorino foggiano cheese. Composition and proteolysis during ripening of Pecorino foggiano cheese (four replicates batches) were analyzed. Proteolysis was greater in cheeses made with rennet pastes from lambs slaughtered at 20 d, as shown by analysis of nitrogen fractions (water-soluble N and proteose peptones). Supplementation of milk substitute with L. acidophilus may have influenced the growth dynamics of lactic acid bacteria in the rennet pastes, with positive effects on levels of lactobacilli in cheese at the beginning of the ripening time. Lower pH values in ARLB cheese during ripening, together with higher cell loads, suggest that supplementation of milk replacer with L. acidophilus resulted in higher proteolytic activity, as also confirmed by the composition of the pH 4.6—insoluble nitrogen fraction. No differences were found in total concentration of free amino acids among the experimental cheeses; phenylalanine, isoleucine, leucine, lysine were found at the highest levels. The addition of probiotic bacteria to milk substitute in lamb rearing appears to give good-quality lamb rennet paste.     

Effect of artisanal kid rennet paste on lipolysis in semi-hard goat cheese

This study examined the use of hygienised kid rennet pastes in model cheese systems and also in goat milk semi-hard cheeses to promote lipolysis. The results obtained indicated that the use of rennet paste caused greater lipolysis and increased, mostly, the short-chain free fatty acid (FFA) content. The model systems made with whole goat’s milk using rennet paste and commercial rennet mixture exhibited a higher FFA content than did the rennet paste-free controls (31,600 vs. 25,600 μmol/kg cheese). For the pilot cheeses made with bovine rennet and rennet paste mixture, the increase in FFA level after 45 days of ripening compared with the cheeses prepared only with commercial rennet was as much as 6600 (μmol/kg cheese) and the increase in the butyric acid content was also 1650 (μmol/kg cheese). Moreover, after 15 days of ripening, industrially prepared cheeses made with rennet paste exhibited greater levels of FFA than did the cheeses made with commercial rennet (11,500 μmol/kg at 45 days of ripening). Their flavour was stronger and the organoleptic characteristics were better accepted, which implies less ripening time for commercial cheese manufacture.     

Proteolysis in model Portuguese cheeses: Effects of rennet and starter culture

To shed further light onto the mechanisms of proteolysis that prevail throughout ripening of Portuguese cheeses, model cheeses were manufactured from bovine milk, following as much as possible traditional manufacture practices – using either animal or plant rennet. The individual role upon proteolysis of two (wild) strains of lactic acid bacteria – viz. Lactococcus lactis and Lactobacillus brevis, which are normally found to high viable numbers in said cheeses, was also considered, either as single or mixed cultures. Our experimental results confirmed the influence of rennet on the proteolysis extent, but not on proteolysis depth. On the other hand, the aforementioned strains clearly improved release of medium- and small-sized peptides, and contributed as well to the free amino acid pool in cheese.     

Evolution of free amino acid content during ripening of Mahon cheese

Evolution of free amino acids (FAA) during ripening (10–300 days) of five batches of Mahon cheese was studied; three batches were made from raw milk and two from pasteurized milk. Major FAA were GLU, VAL, LEU, LYS and PHE (43–67% of total FAA). Relative THR, SER, GLU and GLY contents significantly increased during ripening whereas LEU, PHE and ORN decreased. GLU, ILE, SER, THR, GLY and PHE varied during ripening according to a zero order reaction (r² > 0.99). Principal component analysis showed that the first principal component could be considered representative of the ripening time. The most ripened cheeses showed high GLU, GLY, SER and THR contents and the less ripened cheeses presented high ORN, LEU, PHE and ALA contents. The second component distinguished between cheeses made with pasteurized milk (with high ASN and GLN contents) and raw milk. Indigenous microflora of raw milk showed a strong influence on the proteolytic activity in Mahon cheese.     

Influence of starter culture ratios and warm room treatment on free fatty acid and amino acid in Swiss cheese

Quantification of water-soluble volatile free fatty acids (FFA) and free amino acids (FAA) was performed as a ripening index and an indirect measure of flavor development in Swiss-type cheeses. The objective of this research was to assess the effect of warm room treatment (WRT) and usage ratio of starter cultures, Streptococcus thermophilus and Lactobacillus helveticus vs. propionibacteria, on the concentration of FFA and FAA in pilot plant-scale Swiss cheese. A capillary gas chromatograph equipped with a flame ionization detector was used for the analysis of FFA in Swiss cheese. Free amino acids were analyzed by the Cd-ninhydrin method. Starter culture ratios did not affect development of FAA during the cheese ripening. However, duration of WRT had an effect on the concentration of FAA in the Swiss cheese. Free amino acids increased considerably during WRT. A continuous increase in FAA was shown during 70-d ripening time after WRT. The concentrations of C2:0 and C3:0 fatty acids were affected by starter culture ratios after 2-wk WRT, but these differences had mostly disappeared after 3-wk WRT. Similar concentrations of FFA and FAA reported in previous studies were developed in Swiss cheese with a 3-wk WRT and a 0.33:1 ratio of Streptococcus thermophilus and Lactobacillus helveticus to propionibacteria.     

Effect of NaCl on textural changes and protein and lipid degradation during the ripening stage of sufu,a Chinese fermented soybean food

Sufu is made by solid state fungal fermentation (using Actinomucor elegans) of tofu, followed by salting and maturation in dressing mixtures containing salt, alcohol and various other ingredients. NaCl in dressing mixtures strongly affected the changes in textural properties and the hydrolysis of protein and lipid of sufu. Higher salt contents (14% w/w) resulted in increased hardness (+100%) and elasticity (+18%) and reduced adhesiveness (?30%). Hardness and elasticity could be used to judge the extent of sufu ripening. SDS‐PAGE showed the disappearance of all protein subunits at 80 and 110 g kg^?1 salt content; however, some protein subunits were still detectable at 140 g kg^?1 salt content after 60 days of ripening. Higher ratios of free amino nitrogen to total nitrogen (FAN/TN = 0.4–0.45) and free amino acids to crude protein (FAA/CP = 0.24–0.26) were observed in sufu with lower (80 g kg^?1) salt content. FAN/TN and FAA/CP in white sufu (obtained with dressing mixtures containing only salt and alcohol) were higher than those in red sufu (obtained with dressing mixtures containing angkak or kojic red rice) owing to different dressing mixture compositions. Increases in free fatty acids (FFA) were also observed during ripening. FFA levels in sufu with lower salt content increased rapidly during the first 30–40 days and then increased slowly, probably resulting from the formation of fatty acid esters. Lowering the salt content (80 g kg^?1) can shorten the ripening time to 40 days, which is of benefit to manufacturers. However, sufu will spoil, ie undergo souring, during the ripening stage at salt contents of 50 g kg^?1 or lower. Copyright © 2003 Society of Chemical Industry     

Changes of free amino acid content of Teleme cheese made with different types of milk and culture

The evolution of concentration of free amino acids in Teleme cheese made from sheep, goat or cow milk, using a thermophilic, mesophilic or a mixture of a thermophilic, a mesophilic culture throughout ripening was studied. The total free amino acid (TFAA) content increased at all stages of ripening, regardless of the milk and culture used. In general, the TFAA content was higher in cheeses made from cow’s milk than that of the cheeses made from ewe’s milk; cheese from goat’s milk ranged over intermediate levels. Also, higher concentrations of TFAA were found in cheeses made with the thermophilic than with the mesophilic culture. Cheeses made with the mixture of thermophilic–mesophilic culture ranged over intermediate levels. The results of this study have shown that Leu, Glu, Phe, Val and Lys were the major FAA of Teleme cheese at all stages of ripening, regardless of the type of milk and culture used.     

Free amino acid profile during ripening of ewe's milk cheese from the Croatian island Krk

The aim of this study was to determine the content of free amino acids (FAA) and their ratio in ewe's milk cheese from the island Krk during its ripening. FAA content was determined by reversed phase HPLC (RP‐HPLC) of cheese aqueous/ethanol extracts after FAA were transformed into their 6‐aminoquinolyl‐N‐hydroxysuccinimidyl carbamate derivatives. Their concentration increased during ripening, reaching the value of 5% in cheese dry matter. The dominant FAA were glutamic acid>leucine>valine>aspartic acid>phenylalanine>serine>proline, and higher content of nonessential vs essential FAA was revealed. Krk cheese has, in relation to other cheeses, higher values for glutamic acid/leucine, glutamic acid/phenylalanine, glutamic acid/proline and smaller values for leucine/aspartic acid, valine/aspartic acid, phenylalanine/aspartic acid ratios, while other ratios are comparable to those of other hard ovine 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.     

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.