Determination of the effects of proteolysis-based changes by adjunct lactobacilli on the bioactivity (ACE-inhibitory and antioxidant activities) of cheese: a model cheese study.

Bioactive properties, proteolysis and microbiology of model cheeses with and without adjunct lactobacilli (Lactobacillus helveticus, Lactiplantibacillus plantarum, Lactobacillus bulgaricus and L. casei) were studied during 120 days of storage at 8 or 16 °C. Bioactive properties were observed in peptide fractions (< 3 kDa, 3-10 kDa, < 10 kDa) separated using ultrafiltration membranes. Antioxidant activity of these fractions was determined by radical scavenging assays as ABTS [2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)]. Angiotensin-converting enzyme-inhibitory (ACE-i) activity (% and IC50) and peptide profiles of 70% ethanol-soluble and -insoluble fractions were determined by RP-HPLC. Use of lactobacilli as an adjunct culture significantly changed the RP-HPLC peptide profiles of the cheeses; however, slight changes were observed in the patterns of urea-polyacrylamide gel electrophoresis. Fractions smaller than 3 kDa had higher ACE-i and antioxidant activities for all cheese samples. In conclusion, this study indicates that the addition of lactobacilli as an adjunct culture contributed to the formation of bioactive compounds in the model cheeses and also changed the proteolysis levels.

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.

Changes in volatile composition and sensory properties of Iranian ultrafiltered white cheese as affected by blends of Rhizomucor miehei protease or camel chymosin.

The effect of using various combinations of Rhizomucor miehei protease and camel chymosin (100:0, 75:25, 50:50, 25:75, and 0:100, respectively) on volatile composition and sensory scores in Iranian ultrafiltered white cheese was studied during 90d of ripening. A solid-phase microextraction-gas chromatography-mass spectrometric method was used for determining the volatile compounds of the cheeses. Forty compounds including esters (12), acids (6), ketones (9), alcohols (3), and miscellaneous compounds (10) were identified. The main classes of volatile components in the cheeses are esters, miscellaneous compounds, and ketones. The type and concentration of the coagulants influenced both volatile composition and sensory scores of the cheeses. Principal component analysis separated the cheeses based on the use of 2 coagulants in various combinations and ripening time. The cheeses produced using higher concentrations of R. miehei were separately located on the plot compared with the cheeses produced using higher concentrations of camel chymosin. Sensory evaluation of the cheeses showed that, in general, the cheeses produced using higher concentrations of camel chymosin received higher body and texture and odor and flavor scores than the cheese produced using higher concentrations of R. miehei. In conclusion, 2 combinations of R. miehei and camel chymosin (75:25 and 25:75, respectively) can be successfully used for the production of Iranian ultrafiltered white cheese, considering the results of volatile composition and sensory analysis.

Effect of maceration duration on physicochemical characteristics, organic acid, phenolic compounds and antioxidant activity of red wine from Vitis vinifera L. Karaoglan.

Effects of different maceration times (5, 10 and 15 days) on composition, phenolic compounds and antioxidant activities of red wines made from the Vitis vinifera L. Karaoglan grown in Malatya were investigated. Maceration duration changed some chemical constituents and color of Karaoglan red wines. A linear relationship was observed between antioxidant activity of wine and maceration duration. Major organic acid was tartaric acid which was at the highest concentration in wine macerated for 10 days. A total of 25 phenolic compounds was determined in wine samples. Within these phenolics; procyanidin B2, trans-caftaric acid, gallic acid, trans-caffeic acid, (+) catechin, (-) epicatechin and quercetin-3-O-glucoside were the most abundant phenolics regardless of maceration duration. In general, extended maceration duration resulted in increase in the concentration of phenolic compounds, reflecting the antioxidant activities of wine. In conclusion, the highest concentrations of total and individual phenolic compounds as well as antioxidant activities were found in wines macerated for 15 days.

Study of the chemical composition, proteolysis, volatile compounds, and textural properties of industrial and traditional Beaten (Bieno sirenje) ewe milk cheese.

The objective of this study was to determine the gross composition, proteolysis, and volatile and texture profiles during ripening of industrial (IND) and traditional (TRD) Beaten (Bieno sirenje) cheeses made by using ewe milk. In the course of the analyses, statistical differences were determined in some physicochemical parameters, nitrogen fractions, and total free amino acid levels between TRD and IND types of cheese. Higher levels of proteolysis were observed in IND cheeses than in TRD cheeses during ripening. Levels of residual ß- and αs-caseins were 72.2 and 48.7%, respectively, in 180-d-old TRD cheeses. However, the residual levels were 52.8% for ß-casein and 18% for αs-casein in IND cheeses. Similar differences were noted for the reversed-phase HPLC peptide profiles of 2 types of cheeses. Also, higher concentrations of peptides were eluted in IND cheeses than in TRD cheeses during ripening. A total of 73 volatile compounds, including alcohols (16), esters (17), acids (14), terpenes (7), ketones (5), aldehydes (4), and miscellaneous (10) were identified. The IND cheeses contained higher levels of carboxylic acids, esters, alcohols, and terpenes than the TRD cheeses; however, the same levels of methyl ketones were determined in the 2 types of cheeses at the end of ripening. These may be due to some differences (e.g., pasteurization and scalding temperature, among other factors) in the manufacture of the 2 types of Beaten cheeses. The textural profile of Beaten cheeses showed that TRD production method resulted in firmer, less fracturable, and stiffer cheeses than the IND production method. In conclusion, the results suggest that the use of industrial production method (pasteurization of cheese milk and curd scalding at 70°C) in the manufacture of Beaten ewe milk cheese enriched the volatile profile of the cheese.

Volatiles and sensory evaluation of goat milk cheese Gokceada as affected by goat breeds (Gokceada and Turkish Saanen) and starter culture systems during ripening.

The effect of goat breed and starter culture on volatile composition and sensory scores in goat milk cheese was studied during 90d of ripening. Milk from 2 goat breeds (Gokceada and Turkish Saanen) and different starter culture systems (no starter, mesophilic and thermophilic starters) were used in the manufacture of goat milk cheeses (called Gokceada goat cheese). Volatile composition was determined by a solid-phase microextraction-gas chromatography-mass spectrometric method. Sixty compounds including esters (13), carboxylic acids (7), aldehydes (6), ketones (8), alcohols (14), and miscellaneous compounds (12) were identified. Esters, alcohols, and carboxylic acids were the main classes of volatile components in the cheeses. Both qualitatively and quantitatively, the use of different starter cultures and goat breeds significantly influenced the volatile fraction of goat milk cheese. Decanoic, hexanoic, and octanoic (commonly named capric, caproic, and caprylic) acids were indicator compounds to distinguish the goat breeds. Principal component analysis grouped the cheeses based on the use of starter culture and goat breed. Starter-free cheeses were separately located on the plot and age-related changes were present in all samples. Sensory evaluation of 90-d-old cheeses showed that the cheeses from the Gokceada breed received higher odor, flavor, and quality scores than those from the Turkish Saanen breed, and cheeses made using mesophilic starters resulted in the most satisfactory scores for flavor and quality attributes. In conclusion, goat milk cheeses made using milk from Gokceada goats and mesophilic starter culture had the best quality in terms of volatile composition and sensory attributes.

Acceleration of proteolysis, flavour development and enhanced bioactivity in a model cheese using Kuflu cheese slurry: An optimisation study.

This study aimed to use Kuflu (a mould-ripened cheese) cheese slurry to accelerate ripening, improve biological activity and flavour development in a model cheese in terms of proteolysis and volatile compounds. Response surface methodology (RSM) was employed for the model cheese to determine higher proteolysis and volatile development level during ripening as a function of Kuflu cheese slurry addition level (0-5 %), salt concentration (1-3 %) and ripening temperature (5-15 °C). The highest aminopeptidase activities (0.140 and 0.187 OD/g per hour) were determined in 15-day-old samples containing 3 % and 5 % Kuflu cheese slurry, respectively. Also, the use of Kuflu cheese slurry, regardless of ripening, caused an increase in ABTS*+ antioxidants, angiotensin-converting enzyme (ACE)-inhibition activity and volatile compounds in model cheeses. The sensory evaluation indicated that the use of 3 % (w/w) Kuflu cheese slurry, storage temperature 10 °C for 15 days provided better flavour, odour, texture, colour, appearance and overall acceptability. In conclusion, using Kuflu cheese slurry for model cheese production enhanced the level of proteolysis and volatile flavour composition with shortened ripening time.

Microstructural, textural, and sensory characteristics of probiotic yogurts fortified with sodium calcium caseinate or whey protein concentrate.

The influence of milk protein-based ingredients on the textural characteristics, sensory properties, and microstructure of probiotic yogurt during a refrigerated storage period of 28 d was studied. Milk was fortified with 2% (wt/vol) skim milk powder as control, 2% (wt/vol) sodium calcium caseinate (SCaCN), 2% (wt/vol) whey protein concentrate (WPC) or a blend of 1% (wt/vol) SCaCN and 1% (wt/vol) WPC. A commercial yogurt starter culture and Bifidobacterium lactis Bb12 as probiotic bacteria were used for the production. The fortification with SCaCN improved the firmness and adhesiveness. Higher values of viscosity were also obtained in probiotic yogurts with SCaCN during storage. However, WPC enhanced water-holding capacity more than the caseinate. Addition of SCaCN resulted in a coarse, smooth, and more compact protein network; however, WPC gave finer and bunched structures in the scanning electron microscopy micrographs. The use of SCaCN decreased texture scores in probiotic yogurt; probably due to the lower water-holding capacity and higher syneresis values in the caseinate-added yogurt sample. Therefore, the textural characteristics of probiotic yogurts improved depending on the ingredient variety.

Microbial quality and presence of moulds in Kuflu cheese.

The chemical and microbial qualities, including fungal flora, of 30 samples of Kuflu cheese randomly purchased from different markets in Turkey were investigated. The gross composition of the cheese samples ranged between 37.65-53.65% moisture, 6.21-40.09% fat-in-dry matter, 4.70-10.07% salt-in-moisture and 26.18-44.85% protein. The mean pH value of the cheeses was 6.29+/-0.28 and pH values ranged from 5.52 to 7.22. Variations between the samples in terms of their gross composition suggested a lack of quality standards in cheesemilk, cheesemaking procedure and ripening conditions. The levels of main microbial groups including total mesophilic and coliform bacteria, yeasts and moulds and the presence of some potentially pathogenic microorganisms (E. coli, Salmonella spp. and Staphylococcus aureus) were determined. The high numbers of all microbial groups and presence of potentially pathogenic organisms in the cheese samples suggested that the production and maturation of Kuflu cheese should be improved by better hygiene. Moulds at the cheese surface were isolated and identified. A total of 24 different mould species were detected and the genus most frequently isolated was Penicillium spp. which represented 70.25% of total isolates. Penicillium commune, P. roqueforti and P. verrucosum were the most abundant species in the cheeses sampled. The other dominant fungal groups were Geotrichum candidum, Penicillium expansum and P. chrysogenum. Other genera isolated from the cheese were Acremonium, Alternaria, Aspergillus, Cladosporium, Geotrichum, Mucor, Rhizopus and Trichoderma. The potentially toxigenic species, including some Penicillum spp. and Aspergillus flavus, were also detected.

Microbiology, biochemistry, and volatile composition of Tulum cheese ripened in goat's skin or plastic bags.

Tulum cheeses were manufactured from raw ewe's milk and ripened in goat's skin bags (tulums) or plastic containers to understand the effect of ripening container on the chemical composition, biochemistry, microbiology, and volatile composition of Tulum cheeses during 150 d of ripening. Chemical compositions of the cheeses ripened in tulums were significantly different and the moisture contents decreased rapidly in those cheeses because of the porous structure of the tulum. Higher microbial counts were detected in the cheeses ripened in plastic than in cheeses ripened in tulums. Differences in nitrogenous compounds and total free AA of the cheeses were not significant. Total concentrations of free AA in cheeses increased with age and Glu, Ala, Val, Leu, and Phe were the most abundant AA in the cheeses. Urea-PAGE of pH 4.6-insoluble fractions of the cheeses during ripening showed similar degradation patterns in all cheeses. Peptide profiles by reversed-phase HPLC of pH 4.6- and ethanol-soluble or ethanol-insoluble fractions of the cheeses revealed only minor differences in the concentrations of some peptides among the cheeses; however, age-related changes in peptide concentrations were significantly different among the cheeses. Cheeses were analyzed at 90 d of ripening for volatile compounds by solid-phase microextraction gas chromatography-mass spectrometry. One hundred volatile components were identified, including 11 acids, 16 esters, 12 methyl ketones, 7 aldehydes, 22 alcohols, 7 sulfur compounds, 6 terpenes, and 19 miscellaneous compounds. The main components were short-chain fatty acids, 2-butanone, diacetyl, and primary alcohols. Quantitative differences in several volatile compounds were evident among the cheeses. Cheeses ripened in tulums or plastic had similar aroma patterns, but the concentrations of some components were different.

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 alpha(s1)-casein occurred and that beta-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.