MicroRNAs (miRNAs) are abundant in bovine milk and milk derived from other livestock, and they have functional roles in infants and in the secretion process of mammary glands. However, few studies have evaluated miRNAs in dairy processes, such as during cheese making and ripening. Thus, we investigated the characteristics of milk-derived miRNAs during the manufacturing and ripening of Camembert cheese as well as the microbiota present using the quantitative reverse transcription polymer chain reaction (RT-qPCR) and 16S rRNA pyrosequencing, respectively. Pyrosequencing showed that the cheese microbiota changed dramatically during cheese processing, including during the pasteurization, starter culture, and ripening stages. Our results indicated that the RNA contents per $200mg/200{\mu}l$ of the sample increased significantly during cheese-making and ripening. The inner cheese fractions had higher RNA contents than the surfaces after 12 and 22 days of ripening in a time-dependent manner (21.9 and 13.2 times higher in the inner and surface fractions than raw milk, respectively). We performed a comparative analysis of the miRNAs in each fraction by RT-qPCR. Large amounts of miRNAs (miR-93, miR-106a, miR-130, miR-155, miR-181a, and miR-223) correlated with immune responses and mammary glands were present in aged cheese, with the exception of miR-223, which was not present on the surface. Considerable amounts of miRNAs were also detected in whey, which is usually disposed of during the cheese-making process. Unexpectedly, there were no significant correlations between immune-related miRNAs and the microbial populations during cheese processing. Taken together, these results show that various functional miRNAs are present in cheese during its manufacture and that they are dramatically increased in amount in ripened Camembert cheese, with differences according to depth.
The objective of this study was to determine the physicochemical and sensory properties of Appenzeller cheese supplemented with different concentrations (0, 1, 2, 3, and 4%, w/w) of powdered microcapsules of tomato extracts (PMT) during ripening at 14℃ for 6 mon. The particle sizes of PMT ranged from 1 to 10 m diameter with an average particle size of approximately 2 m. Butyric acid (C4) concentrations of PMT-added Appenzeller cheese were significantly higher than that of the control. Lactic acid bacteria counts in the cheese were not significantly influenced by ripening time from 0 to 6 mon or the concentrations (0-4%, w/w) of PMT. In terms of texture, the hardness of PMT-added Appenzeller cheese was significantly increased compared to the control. The gumminess and chewiness of PMT-added Appenzeller cheese were similar to those of the control. However, both cohesiveness and springiness of PMT-added Appenzeller cheese were slightly decreased. In sensory analysis, bitterness and sourness of Appenzeller cheese were not significantly changed after supplementation of PMT, but sweetness of the cheese was significantly increased after increasing the ripening time from 0 to 6 mon and increasing the concentration from 1 to 4% (w/w). Based on these results, the addition of the concentrations (1-4%, w/w) of PMT to Appenzeller cheese can be used to develop functional Appenzeller cheese.
This study was performed to compare physicochemical and sensory properties of cholesterol-removed Gouda cheese (CRGC) and Gouda cheese made in the laboratory during ripening. Composition, short-chain free fatty acids (SCFFA), texture, color, and sensory properties were measured. In chemical composition analyses, moistures were significantly different between control cheeses (42.86%) and sample cheese (48.32%) (p<0.05). But fat and protein in the control and the sample were 32.77, 22.45 and 31.35, 20.39%, respectively, and were not significantly different (p>0.05). The amount of cholesterol in control was 82.52 mg/100 g and the percentage of cholesterol removal was 90.7%. SCFFA increased gradually during ripening and its level of CRGC increased and significantly different from that of control (p<0.05). The texture, hardness, gumminess, and chewiness were significantly increased, but cohesiveness and springiness were not increased in both cheeses during ripening periods (p>0.05). In comparison of the control and sample cheeses, hardness, and springiness were not significantly different, but cohesiveness, gumminess, and chewiness were different (p<0.05). In color measurement, all color values were not different between CRGC and control (p>0.05). However, $L^*$ value decreased, while $a^*$ and $b^*$ values tended to increase significantly (p<0.05). In sensory properties, appearance, aroma, flavor and taste, and texture were significantly increased except buttery and nutty in aroma and sweetness in taste in both cheeses, and were not significantly different between the control and sample cheeses during ripening (p>0.05). Therefore, this study suggests that the quality of cholesterol-removed Gouda cheese is not different from the control cheese.
The present study was carried out to investigate the changes in chemical and sensory properties, and cholesterol lowering effect of evening primrose oil (EPO) addition in cholesterol-reduced Cheddar cheese. The cholesterol removal rate reached 92.07% by ${\beta}$-cyclodextrin in the cheese before EPO addition. The thiobarbituric acid (TBA) value of cholesterol-reduced and EPO-added cheese increased with both ripening time and amount of EPO addition. Addition of 5% EPO resulted in a significant difference in TBA value after 4-week ripening, compared with no addition of EPO. The production of short-chain free fatty acids (FFAs) increased with ripening period in all treatments. From 4 week of ripening, the amounts of short-chain FFA in 3 and 5% EPO-added groups were significantly higher than those in other groups. Among sensory characteristics, rancidity was mostly affected by EPO addition, however, the rancidity value of 1% EPO-added was not significantly different from that of EPO-free and cholesterol-reduced cheese. Also, Cheddar cheese flavor was not profoundly affected by 1% EPO addition in all ripening periods. Total blood cholesterol dramatically decreased from 184.0 to 137.1 mg/dL with 5% EPO-added and cholesterol-reduced cheese following 8 weeks of feeding. The present results indicated that 5% EPO addition resulted in a profound lowering effect on blood total cholesterol with some adverse effects on chemical and sensory properties.
Objective of the present study is to investigate the effect of coated edible films with chitosan solutions enriched with essential oil (EO) on the chemical, microbial and sensory properties of Kashar cheese during ripening time. Generally, no differences were found in total aerobic mesophilic bacteria, streptococci and lactoccocci counts among cheeses but these microorganism counts increased during 60 and 90 d storage especially in C1 (uncoated sample) as compared with coated samples. Antimicrobial effectiveness of the films against moulds was measured on 30, 60, and 90 d of storage. In addition of fish EO into chitosan edible films samples were showed to affect significantly decreased the moulds (p<0.05) as 1.15 Log CFU/g in C4 (with fish oil (1% w/v) fortified chitosan film) on the 90th d, while in C1 as 3.89 Log CFU/g on the 90th d of ripening. Compared to other cheese samples, C2 (coated with chitosan film) and C4 coated cheese samples revealed higher levels of water-soluble nitrogen and ripening index at the end of storage. C2 coated cheese samples were preferred more by the panellists while C4 coated cheese samples received the lowest scores.
In this study, the physicochemical and sensory properties of Gouda cheese supplemented with microcapsules of chili pepper extract were evaluated. Microcapsules of pepper extract were prepared by coacervation technique using gum acacia-gelatin wall and chili pepper oil core. Changes in pH, lactic acid bacteria (LAB) population, and free amino acid (FAA) content after supplementation of Gouda cheese with chili pepper capsules were monitored during ripening. Texture and sensory characteristics of the Gouda cheese ripened for 6 months were evaluated. The supplementation of pepper extract microcapsules (0.5% or 1%, w/w) did not influence the pH values and LAB content of the Gouda cheese (p<0.05) during the ripening period. While the content of total FAA increased with the ripening process in all the cheese groups (p<0.05), no significant difference (p<0.05) in the content of total FAA was observed among the sample groups at each time point. The addition of pepper extract microcapsules (1%, w/w) to Gouda cheese significantly decreased hardness (p<0.05) and negatively affected sensory attributes in terms of taste and texture (p<0.05). The results demonstrated that supplementation with 0.5% pepper extract microcapsules could provide additional bioactive ingredients, along with maintenance of the quality of Gouda cheese.
Ezine cheese is a non-starter and long-ripened cheese produced in the Mount of Ida region of Canakkale, Turkey, with a protected designation of origin status. Non-starter lactic acid bacteria (NSLAB) have a substantial effect on the quality and final sensorial characteristics of long-ripened cheeses. The dominance of NSLAB can be attributed to their high tolerance to the hostile environment in cheese during ripening relative to many other microbial groups and to its ability to inhibit undesired microorganisms. These qualities promote the microbiological stability of long-ripened cheeses. In this study, 144 samples were collected from three dairies during the ripening period of Ezine cheese. Physicochemical composition and NSLAB identification analyses were performed using both conventional and molecular methods. According to the results of a 16S rRNA gene sequence analysis, 13 different species belonging to seven genera were identified. Enterococcus faecium (38.42%) and E. faecalis (18.94%) were dominant species during the cheese manufacturing process, surviving 12 months of ripening together with Lactobacillus paracasei (13.68%) and Lb. plantarum (11.05%). The results indicate that NSLAB contributes to the microbiological stability of Ezine cheese over 12 months of ripening. The isolation of NSLAB with antimicrobial activity, potential bacteriocin producers, yielded defined collections of natural NSLAB isolates from Ezine cheese that can be used to generate specific starter cultures for the production of Ezine cheese (PDO).
Mucor miehei rennet(MR) was added as calf rennet(CR) substitutes in the fixed amounts of mixed rennets in making Camembert cheese. The conditions in the variations of chemical composition: water-soluble nitrogen, non-caseinic nitrogen, non-proteinic nitrogen, amino nitrogen, ammoniacal nitorgen, electrophoresis, molecular fractionation, mineral distribution, texture characterisitics, free amino acids and free fatty acids, were checked up with the sensory test and the chesse yields at each ripening period. The results obtained by investigating the utility of Mucor rennet were summarized as follows: 1. CR chesse, MR cheese and the mixed-rennet chesse failed to show any significant difference in their yields of 15%. 2. The contents of protein, fat and ash in MR cheese gave lower value than CR cheese did and with progress of ripening lactose decreased rapidly after 14 days of ripening. The difference among the rate of addition of mucor rennet was not recognized. 3. The WSN contents of 5 fresh sample chesse were from 14.7% to 17.3% and WSN increased from 39.7% to 41.0% with progress of ripening. After 21 days of ripening MR chesse had more WSN than CR cheese did. In NCN and ammoniacal nitrogen MR cheese showed higher value. 4. As the ripening progressed, MR chesse showed more cystein, phenylalanine and proline than CR chesse did but it failed to show any increase in aspartic acid, threonine and glutamic acid etc. 5. In the content of free fatty acid MR chesse showed higher value than CR cheese did and with the progress of ripening fatty acids increased from 8.36 mEq to 26.36 mEq but did not show any significant difference in the cheese types by the coagulant ratio. 6. Ca contents in the sample chesse were 0.238-0.27%, Mg 0.019-0.022%, Na 0.910-1.047%, and K 0.175-0.200%. The important non-sedimentable Ca in casein remained from 61 % to 77% without regard the ripening periods and added-rennets and Mg remained from 59.1% to 92.5% in non-sedimentable and water-soluble conditions. 7. In the fractionation of protein by ultrafilteration, MW> $5{\times}10^4$ decresed from 95% at the beginning period of ripening to 45% and MW< $10^4$ increased from 0.2% to 38% and definite caseinolysis was shown in all samples. 8. All the cheese showed to different electrophoretic patterns for the added-amounts of mucor rennet in the 14 days of ripenig. In the 28 days or ripening, MR cheese kept some bands on the patterns compared with CR cheese. 9. In vitro digestibility increased from 81.48-94.81 % to 94.47-98.61% but failed to show any significant difference in the cheese types by the coagulant ratio. 10. In hardness, MR cheese showed lower value compared with CR cheese as the ripening progressed. 11. The results of the sensory test failed to show any difference in flora rind, feelings in mouth and hands, deep structure, flavor and bitterness between CR Camembert cheese and MR Camembert chesse.
For the manufacture of soybean cheese, curds were made from soy milk using either lactic starter or CaCl$_2$and ripened for 3 weeks. When papain was added during ripening, the amout of soluble protein increased. The yields of 3 soybean cheese were 76~88%, indicating that considerable loss of soy protein did not occur during the processing. During the period of ripening the change of moisture content was about 1 n. Total acidity of 3 soy cheese increased, demonstrating the production of lactic acid through lactic fermentation. Texture determined by Instron showed that 3 soy cheese had their own characteristic texture profile curve and that the cheese without papain treatment received the highest textural parameters. 3 manufactured soy cheese received 3-4 (good-very good) sensory scores in appearance, flavor and texture. General acceptibility of 3 cheese were evaluated as desirable or very desirable.
Reactions taken place by a mixed culture of Lactococcus lactis subsp. cremoris KH (lac$^{+}$ prt$^{+}$ ) and KHA (lac$^{-}$ prt$^{-}$ ) and KHA (lac prt ) in cheese ripening have been investigated. Growth characteristics of the mixed culture showed commensalism, and the amounts of proteinases of the mixed culture were small enough. From these results, it is concluded that the production of bitter taste by the mixed culture is a small matter, even if the density of the mixed culture is highly maintained during cheese ripening. Hence, the mixed culture of KH and KHA cells could be a good cheese starter in accelerating the process of cheese ripening.
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