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Food Chemistry 111 (2008) 606–615

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Changes of free amino acid content of Teleme cheese made
with different types of milk and culture
Eleni C. Pappa a,*, Kyriaki Sotirakoglou b
National Agricultural Research Foundation, Dairy Research Institute, 452 21 Katsikas, Ioannina, Greece
Department of Mathematics, Agricultural University of Athens, Iera Odos 75, 118 55 Athens, Greece

a r t i c l e i n f o a b s t r a c t

Article history: The evolution of concentration of free amino acids in Teleme cheese made from sheep, goat or cow milk,
Received 24 October 2007 using a thermophilic, mesophilic or a mixture of a thermophilic, a mesophilic culture throughout ripening
Received in revised form 8 February 2008 was studied. The total free amino acid (TFAA) content increased at all stages of ripening, regardless of the
Accepted 10 April 2008
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 meso-
philic culture. Cheeses made with the mixture of thermophilic–mesophilic culture ranged over interme-
White-brined cheese
diate levels. The results of this study have shown that Leu, Glu, Phe, Val and Lys were the major FAA of
Amino acids Teleme cheese at all stages of ripening, regardless of the type of milk and culture used.
Starter culture Ó 2008 Elsevier Ltd. All rights reserved.

1. Introduction the volatile fraction of cheese (Salles et al., 2000) because they
are precursors of many volatile aroma compounds (e.g. amines,
Proteolysis is the most complex and, in many varieties, the most carboxylic acids, thiols, esters, alcohols, aldehydes and thioesters).
important biochemical event during ripening. The pattern of prote- White-brined cheese, like other types of ripened cheese, needs
olysis can be summarized as follows: the caseins are hydrolyzed maturation to develop the required properties. White-brined
initially by residual coagulant activity retained in the curd and cheeses are in great demand in warm climates and their preserva-
by plasmin and perhaps by other indigenous proteolytic enzymes tion in brine is necessary. Teleme cheese is a soft white- brined
to a range of intermediate-sized peptides, which are hydrolyzed cheese that can be produced using caprine, ovine or bovine milk
by proteinases and peptidases from the starter lactic acid bacteria, alone or any mixture of them (Greek Codex Alimentarius, 1998).
non-starter lactic acid bacteria (NS-LAB) and secondary microflora In Teleme cheese production, traditional yogurt is used as starter
to shorter peptides and amino acids (Upadhyay, McSweeney, by the traditional cheese-makers, while mixtures of commercial
Magboul, & Fox, 2004). Every type of cheese has its own character- starters, including mesophilic starters, are used in industrial
istic free amino acid (FAA) pattern, resulting from the enzymatic production (Anifantakis, 1991). The kind of milk used in cheese
degradation of peptides by various enzymes and also from amino production affects the activity of the starter culture and has a
acid interconversion and degradation (Polo, Ramos, & Sanchez, direct effect on the free amino acid profile of the cheese.
1985). The concentrations of the different amino acids in a cheese There are few publications available on amino acid profile of
are related to the manufacturing technology (type of curd, addition Teleme cheese. Mallatou, Pappa, and Boumba (2004) studied the
of starters, ripening conditions), duration of ripening and the ex- profile of amino acids of Teleme cheese made from sheep’s, goat’s
tent and type of proteolysis (Christensen, Johnson, & Steele, or mixture of sheep’s-goat’s (1:1) milks with mixed thermophilic–
1995). Certain free amino acids (FAA) are extremely important in mesophilic cultures. Buriana and Farag (1983) determined the ami-
flavour and taste development, e.g. Arg is related to bitterness, no acid profile of Teleme cheese made from cow, sheep or mixtures
while Pro, Ser and Asn are related to sweetness (Izco & Torre, (1:2) of these milks with Lactococcus lactis subsp. lactis and Lb.
2000). Although the direct contribution of FAA to the typical casei. Polychroniadou and Vlachos (1979) investigated the amino
cheese flavours are somewhat limited, they are associated with acid profile of Teleme cheese made from a mixture of goat’s-
sheep’s (1:9) milk with thermophilic culture, whereas, Alichanidis,
* Corresponding author. Tel.: +30 26510 94785; fax: +30 26510 92523. Polychroniadou, Tzanetakis, and Vafopoulou (1981) studied the
E-mail address: (E.C. Pappa). amino acid profile of Teleme cheese from deep-frozen curd made

0308-8146/$ - see front matter Ó 2008 Elsevier Ltd. All rights reserved.

70 mM sodium acetate adjusted to pH 6. 11 days. 8 and 11 are gradients programmed by the Waters HPLC con- cheeses made from cow’s milk using thermophilic. standard. Free amino acids were extracted from cheese Friesian cows. Samples were suspended in 100 ll of Pico Tag sample milk was used in making the three variables of cultures and nine diluent (Waters. Phenylisothiocyanate (PITC) (sequential of milk CaCl2 solution (40%.5% acetonitrile. at a quantity of 0. The mixture was homogenized using an Ultra-Turrax dried yogurt thermophilic culture (T). Teleme cheeses 0–3% eluent B over 13. then 9–34% eluent B over 20 min using curve 6. sonicated in a water bath for 20 min and centrifuged at Laboratorium. trol station.4 mM methionine sulfone. The objective of this work was to study the effect of three com.3. USA). Teleme cheese HPLC analyses were done in duplicate.000g for 10 min. holding 24. brated with solvent A for 20 min. and Tarvin (1984). v/v). near the Dairy Research Institute where the as follows: 1 g of cheese was weighed and 10 ml of 0. thermophilic–meso. culture (M). 20 ll of derivatizing solution ganismds was 1:1) and (iii) a freeze-dried concentrated mesophilic (methanol: water: triethylamine: phenylisothiocyanate 7:1:1:1. Analysis of free amino acids (FAA) and chemical analysis mercial concentrated starters. mine: acidic and neutral amino acids: basic amino acids 1:1:1. until both pH and moisture values reached the limits of and 15% methanol. MA.5 min using curve 11.0 g per 100 kg milk. MA. mixed or mesophilic culture. respectively. Gradient elution was as follows: ferred to a cold room (5 °C) for up to 6 months. The samples were incu- the microorganisms Lactococcus lactis subsp. namely a thermophilic. The mixture was left for 10 min in (ii) a freeze-dried concentrated mixture of thermophilic–meso. The solvent system consisted of two eluents made pressure and salting took place in brine (14% NaCl. The (standard solution: internal standard 1:1.mesophilic culture. the milk was standardized to casein to fat (1984). during ripening. respectively. Lb. using thermophilic. whereas. USA). Milford. 0. thermophilus. . HPLC consisting of a controller (model 600).73 for the sheep.5% (v/v). except that glutamine (2. Drainage was under ical grade. That is. goat and cow milk. (Waters. Kandarakis. delbrueckii subsp. and curve 11 maintains starting conditions days. Eluent B was 45% acetonitrile. respectively and afterwards cheeses were trans. injection. profile of Teleme cheese. goat’s or cow’s tives by RP-HPLC using the PICO-Tag amino acid analysis system milk. 13.2. Anifantakis. 40% water (19 °C). con- cheese-makings took place. 3000g for 10 min. E. R-703 (Hansen’s Laboratorium. was manufactured using (a) sheep’s milk of the ‘‘Boutsiko” breed The method of Bidlingmeyer.90  30 cm) was used. cremoris. Before each injection. respectively and 6. bulgaricus. France). an ice bath and then centrifuged at 20. 60. IL. K. Curves 5. The chromatograms were processed using the Millen- 2. Curves 5. cremoris. w/v) was added. 2006a). then 6–9% eluent B over 6 min using philic or mesophilic culture were transferred to the cold room at curve 5. Milford. one of the cultures already men. 2. lactis and Lactococcus bated for 10 min at room temperature and then they were vac- lactis subsp. Run time was 67 min with flow rate 1 ml min 1. at 33 °C. USA) and were filtered through a lots of cheese were obtained. To our knowledge. After cooling. Louis. methionine sulfone and glutamine were purchased from Sigma tioned was added to it.45 lm pore size before day after day basis. too (gluta- ratios of 0.6 and 656%. Rennet powder HA-LA grade) and thiethylamine were from Pierce (Rockford. Methionine sulfone was added as an internal standard and P43% fat in dry matter (Greek Codex Alimentarius. Separations were conducted at 45 °C and the absorbance was mon- The experimental design was that described elsewhere (Pappa.5 min and re-equlibration to 100% eluent A. there Cheeses were sampled for analysis on days 1. lactis. The working amino acid standard was prepared as described in Teleme cheese was manufactured as described previously (Pap.1. cheeses made from at 34% eluent B for 12 min. the deproteinized supernatant were vacuum-dried in the PICO- consisting of the microorganisms Lactococcus lactis subsp. On the day of cheese-making. Briefly. over 10. These results could serve as a model for fu. Each kind of milk was used in making taining 0. a mesophilic or a mixture of a thermophilic. Denmark).1 M HCl. which were vacuum-dried again. on a HV hydrophilic filter (Millipore) with 0. A 2. 500 ll of supernatant were diluted (1:1. until the next step. Identification of the cheese amino acids was achieved by compar- 2. with 40% trichloroacetic acid. & Zerfiridis. acidic and neutral amino acids and cooled at 37 °C. the column was equili- 64. each kind of uum-dried. are no reports of the effect of different cultures on the amino acid as well as on the day they were transferred to the cold room. respectively. 25 ll of philic culture (MX). Pappa. v/v) coccus thermophilus and Lactobacillus delbrueckii subsp. S. as de- from the Agricultural Station of Ioannina (b) goat’s milk from a scribed in the PICO Tag amino acid analysis Operator’s Manual local native goat population and (c) cow’s milk from a herd of (1984) was applied.55 with acetic acid con- After manufacture. 20 ll of a redrying solution (triethylamine: methanol: Lactococcus lactis subsp. consisting of the microorganisms Strepto. 2006a).64 and 0. transferred to the cold room at 22.C. Sotirakoglou / Food Chemistry 111 (2008) 606–615 607 with the same type of milk and culture. respectively. This was repeated five times. 1998). FRC-60 (Hansen’s Laboratorium. standardized milk was pasteurized at 63 °C for 30 min and then Standard mixtures of basic. a solvent pump (mod- el 600E). then followed by a purge at 100% elu- goat’s milk. to obtain a first quality cheese that is 656% moisture v/v/v). (Hansen’s Laboratorium. (the ratio of thermophilic to mesophilic microor. FYS-11 (Rhone-Poulenc blender. Denmark) was added at a The solvents were of HPLC grade. 16 and 12 days. bulgaricus and 1 M sodium acetate 1:2:2. detail in the PICO Tag amino acid analysis Operator’s Manual pa et al.5 min using curve 8. was added as the internal Teleme cheese with three starter culture variables: (i) a freeze. Materials and methods nium 32 software package. on free amino The FAA were identified as phenylthiocarbamyl (PTC)-deriva- acid release of Teleme cheese made from sheep’s. 17 and 15 dients. then 3–6% eluent B made from sheep’s milk using thermophilic. 6 and 8 are convex.. MO. equipped with a 50 ll sample loop. Denmark) consisting of v/v/v/v) were added to the dried samples. v/v/v) were added to the dried samples. Experimental design volume of 20 ll of the amino acids was injected into the column.5 mM) was added. All other reagents were analyt- quantity of 3.75. Tag system. 120 and 180. itored at 254 nm. linear and concave gra- ophilic culture were transferred to the cold room at 23. Samples were analyzed on a WATERS ture research in this field. mixed or mes. a helium degasser and a tunable absorbance detector (model 486). To assist curdling (St. Samples were applied using a rheodyne injector (model 7725i). Horsholm. Cohen. w/w) for 16 h according to the Operator’s Manual (1984). were ent B for 4. cheeses were ripened in the warm room taining 2. Eluent A comprised at 19 °C. Cheese-making and sampling ing retention times with pure standards. USA). A Waters reverse phase column (3.

2 AAB and b-Ala were for selecting discriminant variables. The third principal component explained another 10. Pappa.2. 4 values greater than 1. Fig. Sotirakoglou / Food Chemistry 111 (2008) 606–615 The dry matter (DM) of the cheese samples was determined Biplot according to the IDF standard (IDF. One-way analysis of variance was used for the -9 -6 -3 0 3 6 comparison of the data of each parameter of Teleme cheese made Component 1 (30. 1. Cheeses made with thermophilic culture were clustered near (SPSS for Windows version 14. at different ages. sented the opposite characteristics.85% of the total variability and fied peaks. cipal component could be a representative of the type of culture. -2 Tau They were away from the axis origin. were co-injected with the standard mixture solution to identify peaks in the standard solution. because all cheeses. Consequently. the third princi- During the extraction and derivatization process. Gly and Lys. was made. while -6 -4 -2 0 2 4 6 cheeses 120 to 180 days old were placed on the negative side of Component 2 (10. 1). together with the intact FAA.. However. On the other hand. these cheeses had the lowest content of Ser.. a number of pal component could be a representative of the kind of milk. goat’s or cow’s) as factors and ripening time as Asn Leu -2 split–splot factor. Leu Arg Lys Ile Val Orn indicating that they were positively correlated with each other. mixed thermophilic–mesophilic or mesophilic).0.C.1. and was Met Asn Gly Trp Glu 0 TFAA mainly defined by TFAA. Plot of the free amino acids as a function of the two first principal components. at each -6 age. San Diego. than cheeses made with the other cultures. goat’s or cow’s) with the three types of culture (thermo- philic. ing opposite to Gaba.4. Moreover. On the other hand. Consequently the second prin- USA). which repre- ville. Rock- acids. therefore they were negatively correlated to Statistical analysis was performed using statistical packages this. In Fig. Principal component analysis.42%) 2 Asp AAB ability. General had the highest contents of Pro and Ala (Fig. IL. Ser B_Ala Component 3 (10. 1 are ripened 180 d old cheeses. The second principal component explained the second and third principal components. CA. Principal component analysis (PCA) was applied to pooled data in order to reduce the dimensionality of the data and to detect the most important causes of variability. Gly and Lys. the type of 10. Cheeses made from goat’s milk were clus- tered on the negative side of PC3. which could be considered as Ala Pro representative of the ripening time. The eight selected components accounted for 79.68%) PC1 (Fig.75% of the total variability. these amino acids were ly- time. STAT- AAB and b-Ala and therefore had higher contents of these amino GRAPHICS Plus for Windows version 5.0. SPSS Inc. K. considering the type of culture (thermophilic. generally. a comparison of the values of each Cit Met parameter of Teleme cheese manufactured with one type of milk -4 Gaba (sheep’s. free amino acids were represented as a function Cit of both first and second principal components. Chicago. PCA of the 27 free amino acids resulted in eight principal components with eigen. B_Ala AAB 4 Component 2 (10.75%) with one type of milk and one type of culture. a common statistical cut-off point.608 E. Cheeses made from Amino acids were identified according to their retention times sheep’s or cow’s milk had the opposite characteristics and were by comparison with a standard mixture solution chromatogram. having the highest contents of TFAA. USA.05). . Results and discussion and Ser (positive value). 2. indicating a high were applied to fit the evolution of free amino acids with ripening positive correlation. Manugistics Inc. All the cheeses that were located on the left of Fig.8. The least significant differences were obtained using the LSD test Fig. These amino acids were Gln Phe His placed closed together on the negative side of the horizontal axis. Ala (negative values) 3. Asn. 0 GlyPhe Orn mixed thermophilic–mesophilic or mesophilic) and the kind of Gln Arg Tyr milk (sheep’s.42% of the total variability and was defined by Pro. from 1 -4 to 60 days old.68% of the total variability and was defined by AAB. Principal component analysis. In Fig. these peaks did not interfere with the identi- together they accounted for 51. and the results were ex- 6 pressed on a dry matter basis. TFAA AspThrHis Tau AlaCys Lys Ser plied to the data. Statistical analysis Val Ile Glu Trp 2 Pro A three-way split–splot analysis of variance (ANOVA) was ap. 1986). USA and GraphPad Prism version 4. Plot of the free amino acids as a function of Asn. since a great correlation be- Biplot tween the free amino acid contents was noticed. The first principal Cys Thr Gaba Tyr component explained 30. 2). b-Ala (posi- milk and the stage of ripening. (P < 0. Wilk’s lambda (k) criterion was used tive values) and Gaba (negative value).14% of the total vari. first three principal components were the most important. were clustered on the positive side of PC1. near Pro and Ala and therefore 3. Linear regression analyses placed close together on the positive side of PC2. When necessary. suggesting that they were well represented from the first PC. standard solutions of pure amino acids had eigenvalues greater than 2.68%) 2. MD. Principal component analysis (PCA) was used to reduce the dimensionality of the data and stepwise discriminant analysis was also applied to establish those amino acids capable of discrim- inating and classifying cheeses by the type of starter. The unidentified peaks were present. placed on the positive side of PC3. 1.

914 *** P < 0.996 *** 0.620 *** 0.995 *** 0.5% of the TFFA. Franco.868 *** 0.908 *** 0.958 *** 0.983 *** 0. and Ordonez (1995) reported an increase from et al. Phe.996 *** 0.001.550 *** 0. The proteolysis of as1-casein. Guven.974 ** 0.960 *** 0.960 *** 0.952 Bala *** 0. Effect size Sig.598 *** 0. and Sotirakoglou (2006b) reported an 200 mg 100 g 1 DM in 1 day old to 1300 mg 100 g 1 DM in Idiaza- increase of proteolysis during cheese maturation. Fox.740 Arg *** 0.872 *** 0.880 *** 0.908 *** 0.966 *** 0.01.971 *** 0. cata.993 *** 0.988 *** 0. K.940 *** 0. kind of milk (M).982 *** 0.970 Val *** 0.923 *** 0. The increase of TFFA at all stages of ripening. & Xavier Mal- study.989 *** 0.577 *** 0. in Cheddar cheese 1592 mg 100 g 1 DM. Effect size Sig. Barcina ies.05. weeney. which has a high content Table 1 Significance levels and size effect of the three-way split–splot ANOVA for the effects.900 Thr *** 0.997 *** 0.985 *** 0. Buriana and Farag (1983) found that the TFFA decreased From Table 2 and Fig. . Val and after 180 days of ripening and re-increased rapidly until day 360. Pappa. Javier Carballo.984 *** 0. regardless of the type of milk and culture used. 2005.937 *** 0.925 *** 0. Effect size Sig. 400 ening time (P > 0.953 *** 0.981 0.824 Cit *** 0.926 *** 0. in tent. to 424 mg 100 g 1 DM in 60 day old cheese. increase of hydrophilic peptides and decrease of hydrophobic Edam cheese 1056 mg 100 g 1 DM and in Romano cheese peptides.991 *** 0.965 *** 0.994 *** 0.961 *** 0. 0 60 120 180 vidual FAA of Teleme cheese made with different starter cultures Ripening day and milks during ripening. kind of milk.994 *** 0.493 *** 0.001) for the total free amino acids (TFAA) of Teleme cheese and for all of the individual free ami- no acids. & McS- found in this study. The FAA in Manchego cheese typically contribute the increase of nitrogen faction. The levels of TFFA Fox.909 *** 0.949 *** 0.934 *** 0.984 *** 0. in this 650 mg 100 g 1 DM (Tavaria.971 *** 0.938 *** 0.985 *** 0.423 *** 0.979 *** 0.05).829 *** 0. 2003).958 *** 0.979 *** 0.985 *** 0.996 *** 0.940 *** 0.001).905 *** 0. Table 1 also shows the effect size (>0.964 *** 0.736 *** 0.964 *** 0.966 *** 0. Lys were the major FAA of Teleme cheese at all stages of ripening.921 Gaba *** 0.957 *** 0.939 *** 0.896 *** 0.952 *** 0. Hayaloglou.934 ** 0. Kand- arakis.823 *** 0. 3.897 *** 0. Generally. Glu. In Table 2 it can be seen that the con- Fig.980 *** 0.986 *** 0.985 *** 0.875 *** 0.863 *** 0.984 *** 0.983 *** 0.940 *** 0.913 Glu *** 0. Evolution of total free amino acids during the ripening time.928 *** 0.965 NS 0. is in accordance with the results found by Mallatou et al.732 *** 0. Fig.634 *** 0.894 *** 0. which was not significantly different with respect to starter addition (P > 0. Ibanez.971 *** 0.948 *** 0.05) 200 for the TFAA and for all individual free amino acids. **P < 0. except Ala.934 Leu *** 0.811 *** 0.861 * 0.866 *** 0. 3 presents the evolution of mean total free amino acids (TFAA) of Teleme cheese during ripening.2. type of culture and rip.990 *** 0.943 *** 0.984 Tyr *** 0.990 *** 0. centrations of almost all FAA showed a clear tendency to increase with ripening time. & McSweeney. Pappa. Effect size AAB *** 0.912 *** 0. as was expected since.973 *** 0.966 *** 0.959 *** 0.970 *** 0.901 *** 0. Effect size Sig. Guven.982 *** 0.932 *** 0.989 *** 0.984 Phe *** 0. 4 it can be seen that Leu.988 *** 0.986 *** 0.978 *** 0. Anifantakis. which did not change significantly during rip.994 *** 0.05) and Tau. regardless of the type 316 mg 100 g 1 DM on the day cheeses were transferred to the of milk and culture used.869 *** 0. *P < 0. are within the range reported from other stud.966 *** 0.824 *** 0. TFFA mean content of Teleme cheese increased sig..796 *** 0.939 *** 0. Barcina. Vicente.938 *** 0.991 Cys *** 0.911 *** 0. 2001.993 *** 0. were significant (P < 0. these compounds are released by proteolytic agents. Ibanez.078 *** 0.931 Gln *** 0. Barcina.903 *** 0.949 Met *** 0. 2004.742 * 0.974 *** 0.939 *** 0.951 *** 0. to in Teleme cheese by Mallatou et al.994 *** 0. At the age of 180 nificantly from 295 mg 100 g 1 DM in 1 day old cheese to days.C. NS: not significant. Table 2 shows the concentrations of indi. Effect size Sig.965 *** 0.980 *** 0.882 *** 0.977 *** 0.973 TFFA *** 0.988 *** 0.936 *** 0.964 *** 0.886 *** 0.993 *** 0.993 *** 0. Hannon.993 *** 0.990 * 0. as estimated by bal cheese.833 Asn *** 0.590 *** 0.989 *** 0. they accounted for 41. The same FAA were found in abundance cold room.915 *** 0. during proteolysis.946 *** 0.975 *** 0.883 *** 0.979 *** 0.668 ** 0.930 Ser *** 0.8 in most cases).962 *** 0.990 *** 0.990 *** 0. Effect size Sig. 100 g 1 DM in180 day old cheese (P < 0.910 NS 0. (2004).953 Gly *** 0. (2004) and Polychroniadou and 492 mg 100 g 1 DM in 120 day old cheese and finally to 675 mg Vlachos (1979).913 *** 0.970 *** 0. decrease of the intact casein con.907 *** 0.938 Ala *** 0.986 *** 0.942 *** 0. Zerfiridis.980 *** 0. E.806 *** 0.839 His *** 0.434 *** 0. All two-way and three- way interactions between the factors were significant (P < 0. & Barron. which describes the pro- 0 portion of total variability attributable to a factor.509 *** 0. 1995).912 *** 0.954 Asp *** 0.902 *** 0.944 *** 0.867 *** 0. 600 ening time. type of culture (C) and ripening time (T) Amino acids M C T MC MT CT MCT Sig.900 Pro *** 0.980 Ile *** 0. Sotirakoglou / Food Chemistry 111 (2008) 606–615 609 3.849 *** 0. Effect of ripening time 800 Total amino acids mg/100g DM The results of the three-way split–splot ANOVA (Table 1) showed that the main effects.916 Trp *** 0.852 *** 0.990 *** 0.930 Orn *** 0.991 *** 0.926 *** 0.988 *** 0.903 *** 0. as well as in other cheeses (Hayaloglou.962 Tau *** 0.925 *** 0. 1728 mg 100 g 1 DM.860 *** 0.967 Lys *** 0.683 *** 0.

82b Thr 5.62a 2.93b 0.82a 12.02b 7.31b 5.83a 22.61ab 9.21a 0.82a 5.312a 7.21a 14. (days) 1 RB Milk S G C S G C Culture T MX M T MX M T MX M T MX M T MX M T MX M Asp 1.41b 12.81a 8.21b 4.42a 8.12b 4.92a 44.02a 11.71a 11.812ab 12.91a 6.32b 17.72a 38.11a 2.41a 2.62a 40.02ab 16.12a 11.11a 18.712a 0.42a 4.02b 0.81b 1.71c 8.71a 0.92a 26.81b 24.82a 5.91a 5.31a 3.01b 5.41a 21.62a 29.92a 7.82a 0.61b 5.22a 8.21b 11.93b 1.42b 4.52a 7.92a b-Ala 16.91b 7.51b 19.22a 0.53b 55.21a 29.42a 3.312c 11.92a 7.92a 2.12a 10. goat’s (G) or cow’s (C) milk using a thermophilic (T).21c 6.12a 7.91a 2. .41b 2.81ab 19.112a 3.33b 0.31ab 1.01a 1.22a 0. a–c : For each age and each kind of culture.42a 0.81a 4.02a 0.82b 8.32b Cit 0.42b 7.31a 8.12c 8.21a 2.31b 3.82a 17.01a 4.03a 20.51c Tyr 11.93a 19.52b 3.72b 1.93a 21.51a 0.22a 11.21a 6. a mesophilic (M) or a mixture of thermophilic and mesophilic (MX) starter during ripening Age.21a 3.91b 1.61a 3.41a 52.11b 4.92a 3.02a 1.61a 1.02b 0.112c 19.91a 0.41a 16. 610 Table 2a 1 Changes of amino acid contents (mg 100 g DMA) of Teleme cheese made from sheep’s (S).52b 21.02a 10.91a 73.52a 15.12ab 5.41a 1.52a 46.012a 1.51b 24.91b Asn 12.01a 0.51a 2.12b 11.52a 8.61a 13.22a 1.32b 22.31b 5.12a 1.02b 0.02a 0.02b 2.13a 19.91a 0.32a Arg 27.71a 0.71a 13.91a 16.41c 22.23a 51.81a Orn 14. different letters indicate significant differences (P < 0.02a 9.91a 13.12a 12.52a 9.02ab 13.81c 2.81a 0.01a 8.81b 5.51b 8.01c 4.42a 21.81b 4.72a 34.82a 9.31c 37.912a 23.81a 3.11b 38.12a 6.32b Ala 3.43c Gaba 3.93a 28.81c 8.21a 37.63c 18.91b 16.42a 1.31a 3.81a 4.61b 7.41b 3.11a 6.01a 0.11 2.21b 5.32b 7.11a 2.52a 3.81c 5.41b 23.712a 8.71b 0.81a 0.22b Gln 7.51b 16.22a 15.91b 11.12a 1.01a 6.81a 0. K.52b 33.93b Val 16.52a 11.03b 0.03b 84.21c 3.52a 35.41a 25.43c Cys 6.62a 2.61a 14.12a 32.33a 2.52b 7.61a 1.63b 15.72b 2.12a 1.81a 6.22a 10.21b 5.52a 3.71a 5.81a 25.21a 8.42ab 7.81a 8.71a 10.63a 17.22a 11.13a 29.22a 9.01b 0.05) between kinds of milk.81b 1.81a 17.12b 1–3 : For each age and each kind of milk.71a E.92a 16.33a 6.32a 11.52b Trp 2.22a 0.52b 3.02a 2.91b 4.02a 0.72c 4.41a 4.11b 6.33c 0.62b 0.01a 17.52a 0.52a 1.23a 7.93b 8.71a 5.71a 1.71a 10.71ab 13.42a 31.31a 6.92a 15.91a 2.72a 0.21b 1.53a 34.01a 8.81b 7.11a 3.72c 36.71b 28.01b 6.81b 6.31c 2.41b 13.61a 14.61b 3.412b 25.33b 56.71b Leu 23.712a 2.11a 1.512a 26.412a 10.91b 14.51a 8. Sotirakoglou / Food Chemistry 111 (2008) 606–615 Glu 30.11b 1.71a 14.81a 6.42a 5.02a 6.83a 3.01a 3. B R: Day cheeses were transferred to cold room.32b 8.11a 11.51a 9.42a 3.05) between kinds of cultures.51a 13.11b 24. Pappa.91b 2.02b 58.21a 3.51ab 6.212a 3.61a 1.21b 3.02b 2.12a 26.81a 43.41a 1.91a 0.72b 3.01c 37.72a 0.81a 9.31a 1.13c 67.71b 23.51a 15.01b 16.73b 17.02b 1.62a 35.82b AAB 26.11b 0.22a 5.92a 1.72a 0.62a 7.81b 7.52a 3.71a 3.12a 5.11b 2.21b 9.61a 1.02a 9.21a 10.82a 7.43c Ser 3.72a 6.312b 15.01b 28.11a 11.02a 36.11b 2.21a 13.43b 2.21b 13.412b 4.21b 7.21a 13.11b 1.11b 17.12a 23.81b 26.12ab 0.61a 11.11a 35.62a 6.212a 32.21b 3.93a 9.81b 3.82c 2.01b 5.82a 6.41a 1.71b 5.31a 49.01a 0.71b Met 18. %.12b 12.11b 10.72a 35.01a 6.01c 3.81b 12.01b Ile 40.22a 3.42a 20.02a 15.71a 6.12a 3.02a 4.71a 6.92c 1.31b 10.32b 23.52a 2.81a 0.81b 17.42a 0.01b 14.53c Phe 22.42b 11.11b 31. A DM: Dry Matter.71b Lys 66.212b 14.31a 6.81a 1.92a 16.61a 6.22a 1.02c 4.62a 0.21a 3.92b 5.21a 1.52a 64.02a 20.81b 19.11b 19.02a 8.01a 1.61a 4.51b 3.31b 23.62ab 2.81a 9.42a 10.91a 3.92a 8.21a 1.81b 68.91b 3.01a 2.92b 33.22a 2.71a 1. different numbers indicate significant differences (P < 0.61b 6.92a 13.91b 1.02b 2.12b Gly 6.21b 7.12c Tau 2.32a 16.01a His 4.01b 38.81b 15.22a 3.C.82b 3.22a 63.22a 84.82c 0.73a 43.81b 12.81a 9.32a 16.11b 22.51a 12.31a 1.02a 1.32a 34.31a 0.21b Pro 7.33a 19.91b 14.81ab 2.31b 16.33a 23.41b 7.72a 4.51a 2.71a 3.02a 4.52a 0.11b 16.02a 11.11a 29.71a 33.52a 32.92b 18.61b 2.01a 2.12a 4.72b 13.12c 5.

212a 26.01a 78.51a 1.91a 4.22a 3.31a 8.01a 8.62a 13.31a 6.71a 4.11a 30.71a 19.61b 8.21a 18.91a 18.02a 5.93a 1–3 : For each age and each kind of milk.12a 2.91b 10.91a 3.41a Asn 5.81b 19.42a 42. a–c : For each age and each kind of culture.41a 13.71a 6.83a 22.71a 25.92a 9.51b 15.11b 11.51b 77.03a 0.41a 5.31a 0.01a 0.81a 4.62a 11.12b 8.11c 12.91a 4.22a 4.52a 6.82a 7.91b 11.52a 26.01b 25.41a 5.01a 3.41b 34.21a 24.62c 59.61a 6.91b 4.51a 13.31a 4.81a 11.63a 28.52a 8.11b 6.41a 19.01a 52.71a 28.01a 27. Sotirakoglou / Food Chemistry 111 (2008) 606–615 Asp 5.12b 2.112a 15.31a 8.05) between kinds of milk.01b 2.51b 1.42a 9.41b 10.81a 5.51a 11.82a 37.41a 0.22a 6.21b 14.11b 1.31a 13.41b 13.92a 6.61a 0.41b 6.61a 6.512a 10.62a 13.41a 3.312a 14.21a 40.41a 20.21a 15.22a 5.22b Phe 67.32a 0.01a 23.61a 37.61a 2.21b 34.31a 3.31a 4.03a 12.31a 3.02b 22.91b 17.12b 8.22b 12.21a 8.11a Thr 15.41a 31.05) between kinds of cultures.81a 1.41a 16.81c 40.22c 61.52a 36.81a 0.91a 48.62b 23.42a 13.01c 84.42a 12.91c 15.82c 6.51a 5. %.12b 19.73a 31.52a 38.52a 11.52a 30.62c 9.22a 6.91a 0.11a 51.31b 27.81a 2.41a 0.912a 1.21a 20.91b 5.91a 4.31a 13.23b Gaba 3.11a 81.82b 20.01ab 3.92ab 0.31a 2.42a 19.61b 3.11b 3.01b 5.41b 2.61b 0.51a 44.21b 33.51b 10.01a 11.92b 16.82a 1.52a 10.81a 1.02a 5.81b 36.81a 7.71a 51.21a 4.92a 12.72b 30.01a 1.71a 0.22a 4.01b 7.81b 6.42a 1.01a 6.61b 26.91b 6.51a 9.41a AAB 46.02a 3.73a 43.81a 3.53a 35.32a 2.23c 46.02b 17.02b Ser 5.32ab 21.Table 2b 1 Changes of amino acids content (mg 100 g DMA) of Teleme cheese made from sheep’s (S).02ab 10.02a 71.52a 8.71a 8.12c 7.81b Pro 8.11a 4.11b 135.41a 16.93a 37.53b Ile 35.412a 2.31a 9.52a 0.91a 0.42b 16.72b 6.02b 1.11b 33. a mesophilic (M) or a mixture of thermophilic and mesophilic (MX) starter during ripening Age.02a 13.01a 2. K.72b 21.32b 67.01a 3.31a 0.92a 20.51a 2. 60 120 (days) Milk S G C S G C Culture T MX M T MX M T MX M T MX M T MX M T MX M E. A DM: Dry Matter.62a 6.11b 7.01b 22.82a 7.41a 28.73ab 4.11a 9.61b 3.21a 5.72c 11.41a 26.73b 48.93b 17.72c 3.52b 4.42a 8.72b 1.92b 5.72a 14.62b 0.22a 20.82a 18.83b 8.91b 16.93a 140.21a 10.61a 8.03b 9.61b 15.71a 5.91a 91.91a 12.63a 103.02b 52. Pappa.11a 3.312a 16.81b Trp 7.21a 3.12b 0.62b 1.21b 2.52a 28.03a 43.71a 3.81b Gly 5.11a 21.52b 12.82a 6.11a 2.11b 0. goat’s (G) or cow’s (C) milk using a thermophilic (T).81b 5.91b 14.12b 11.31b 14.61c b-Ala 23.63b 22.42b 13.33b 5.91a 3.32a 3.12a 12.71a 3.11a 12.21ab 7.32a 7.41b 9.92a 21.02a 23.11c Leu 16.01b 101.31a 20.61c 2.5a Arg 29.53b 3.82b 6.12a 12.91b 20.51b 1.42b 58.61a 7.51a Gln 8.91b 1.812b 3.31a 17.01b 77.11a 11. 611 .C.62c Met 20.42ab 0.21a 29.73b 3.31a 11.72a 12.22a 3.31a 4.01c 147.72a 2.63ab 9.01b 9.71c 38.11a 1.13b 3.21b 9.91b 17.51a 1.82b 26.73b 4.62b 9.03a 9.71b 46.52a 2.02b 0.71a 21.41b 11.12a 0.41a 4.72a 1.33b Orn 7.61b 19.11a 20.412a 132.21a 3.31a 38.02a 5.31a 49.412b 25.03a 41.11a 1.61a 16.82c Val 28.82a Tau 3.12a 12.72a 7.51b 34.23b 4.01a 4.32a 1.71a 3.11b 4.91b 157.012a 64.01b 5.82b 2.02b 1.12b 12.31a 4.42a 18.71c 7.61a 15.72c 3.31b 13.912a Tyr 6.71b Ala 4.01ab 3.02c 10.42c 10.01a 28.41a 40.22a 15. different letters indicate significant differences (P < 0.21ab 13.512a 7.12b 6.23c 70.41a 23.12a 1.02b 9.62a 3.12a 13.91a 14.92a 3.72a 72.81b 17.91a 24.42a 6.43b Glu 122.52b 4.81ab 8.12a 134.91a 2.41b 3.22b 3.22a 1.22a 4.012a 22.81a 3.92b 40.51b 0.01a 38.51b 15.41a 4.42b Cit 3.01b 11.41a 8.42a 38.02a 10.91b 3.42a 36.71a 4.12b 3.43b 18.12b 34.32b 8.51a His 13.12b 4.91a 7.01a 30.22b 22.93a 31.21c 24.72a 5.11a 2.11a 11.72b 48.03c Lys 59.72ab 23.61a 18. different numbers indicate significant differences (P < 0.92a 3.92b 3.11a 6.92a 32.53b Cys 6.22b 23.12b 3.62c 2.01a 2.41c 41.62a 0.81a 2.51a 9.11a 4.11a 5.32b 16.42b 3.22b 5.71a 38.01a 1.91a 11.62a 0.62b 51.11a 1.41b 37.02c 114.71b 18.61a 30.51b 0.21a 3.01a 3.91b 5.51a 5.51a 11.52a 27.02a 0.51c 28.52b 32.83c 66.91b 30.

12a 1.51a 4. centroids corresponding to R.72a 4.92a 6.51b 0.01a 17.52a 27.01b 9.71c Pro 34.91a 5. often the samples that were classified into the correct group by the stage a-ketoglutarate.22a 1.52a 62.01a 89.21a 13.93a 9.11c 8.01a 5.11b 23. Some overlapping between R. different numbers indicate significant differences (P < 0.12a 5.11b Arg 39. Leu and Met contributed mainly to discriminant function D2 Teleme cheese samples had high contents of Glu (Fig.51b 19.12b 23.01b 6.22a 42.51b 7.41b 44.32a Leu 36. Asn of the amino acids Leu.6 mg 0 100 g 1 DM in 180 day old cheese).51b 3.22a 59.22a 5.91b 19.41a 39.62c Gaba 1.03c 1–3 : For each age and each kind of milk.41a 50.81b 7.51a 6.52a 34.612 E. (days) 180 Milk S G C Culture T MX M T MX M T MX M Asp 15.81a 0.92ab Ala 4.62c 23.01b 97.41ab 11.72a 3.42b 52.412a 0.92a 26.01c 3.23a 9.71a 2.63b 15.23a 7.21a 22.91b 7. 4).91a 3.81c 34.61a 20.. Free amino acid mean content of Teleme cheese.32a 17. is transformed to the corresponding amino acid of ripening was 80.8 mg 100 g 1 DM in 1day old cheese to 11.32b 12. a mesophilic (M) or a mixture of thermophilic and mesophilic (MX) starter during ripening Age.53a 18.112a 46.05) between kinds of milk.71c Orn 14.32a 27.41c 92.05).61b Gly 14.82b 19.71a 1.92b 72.02a 16.81b 5.01c 112. 1993).51a 4.51b 30.91c 36.21b 67.32b 8.05) between kinds of cultures.72b 7.312 16.23b 132. Four dis- criminant functions were statistically significant (P < 0.11c 14.32a 14. 5).02b 0.92c 11.72c 9.12b 1.53a 27. therefore it is important that this amino acid concen- 30 tration does not increase with ripening time.02a 7.02a 20.62b 1.81a 8. 4.02b Glu 86. occurs during the first days of and Tau contributed the most to discrininant function D1(54%). 50 The mean concentration of Arg was 14.62a 3.42b 28.0%.43a 31.92a 9. different letters indicate significant differences (P < 0. The distances between the group with cheese ripening was found for AAB.42b 66. ripening.62a 6.61a 53.92a 60.31a 25. K. Stepwise discriminant analysis selected only nine amino acids Fig.23b 5.22a 15.01b 74.01b 34. its content represented less AAB ALA ARG ASN ASP BALA CIT CYS GABA GLN GLU GLY HIS ILE LEU LYS MET ORN PHE PRO SER TAU THR TRP TYR VAL than 2% of the TFAA content.62a 9.62c Trp 7.82c Met 10. Gaba (which is produced from Glu by a decarboxylase and is 20 present in low-quality cheeses) was found at low concentrations in Teleme cheese.83b 30.12c 7.82c 26.13c Ser 16.51a 7. The low concentration of Pro in Teleme cheeses could be 60 because this amino acid is normally released as a dipeptide (Prit- chard & Coolbear.11b 0.71a 23.21a 5.23a 29. %.72c 32.41a 18. Phe and Val. for discriminating among the stages of ripening (Fig.42a 5.712c Phe 38.62a 14.03a 15.61a 16.72b b-Ala 25. 60 and 120 day old samples were Pro mean concentration was 13.23c Asn 49.61b 36.61b 41.212b 45. Cys and Ala.2 mg 100 g 1 DM in the 1 day old cheese and reached 26.02a 34.31b 14.41b 6.61c 57.72a 0.41a 22.61b 3. Pappa.71a 43.82b 37.51a 42.33c Val 94.92a 6.12b 81.02b 29.71a 3.01a 70.91b 2.81a 0.12a His 52.41a 3. 60 Glu (Yvon & Rijnen.02 23.2 mg 100 g 1 DM in the 1 day lower than the distances corresponding to 1 and 180 day old old cheese and increased to 24.61a 5.41a 16.52b 81.66%) and Orn acid transamination is catalyzed by aminotranferases and results and Leu to discrininant function D4 (2.12b 4. 5).21a 23.92b Tyr 31. Arg is responsible for unpleasant or bitter taste (Polo et al.42a 38.62 34.52b 15.41a 5.32a 1. A DM: Dry Matter.41a 3.92a 5.42b 30.11a 30.03a 112.92b 60. cheese. (Fig.02a 108. A similar decrease may not be completely separated.52a 63.53a 105. Amino (35%). The percentage of in the formation of a-ketoacids while the a-ketoacidacceptor.33a 63.82a 79. goat’s (G) or cow’s (C) milk using a thermophilic (T).61b 1. .51c 29.72b 4. a–c : For each age and each kind of culture.12b 25. In contrast to TFAA and most of the and 120 day old samples was observed.912b Tau 6.212a 2.61b 48.0 mg 100 g 1 DM in 180 day old mg/100g DM 40 cheese. Despite the fact that the mean concentration 10 of this amino acid increased continuously throughout the ripening period (from 4.3 mg 100 g 1 DM in 180 day old samples.22a 5.42c Cit 33. As ripening is a continuous process.62c 6.31a 23.21a 21.02a 7.02a 14.02b 56.81a 7.01b 53.212b AAB 34.11b 1.22a 34. 1985).21a Ile 34.41b 28.08%).C.41a 36.01b 2.52a 7. the groups presented a maximum around day 60 or 120.01c 105.03a 56.03a 7. which predominates over the proteolysis of b-casein.01b 9.21c 16.62a 5.11a 2.11b 61.51a 17.41b 12.91a 14.51a 33.22ab 8.42a 46.02a 24. Sotirakoglou / Food Chemistry 111 (2008) 606–615 Table 2c Changes of amino acids content (mg 100 g 1 DMA) of Teleme cheese made from sheep’s (S).82a 7.83b Lys 150.12c 10.01b 31. which caused the mis- other free amino acids.01a 8.612b Gln 32.31b 24.42b Thr 31.71b 24.11a 38.82a 21.72c 7.52b 5. 2001). Glu and Leu to discriminant function D3 (7.512b 1.21c 0. Glu decreased with cheese ripening and classification. at 180 days of ripening.31a 4.11c Cys 5.93a 63.62a 9.42b 5.31b 5.63b 25.31b 46.

Both D1 and D2 were significant (P < 0.. Orn (as well as Cit. 6 shows the evolution of TFAA for the three types of milk possesses two additional peptidases (an oligopeptidase and amino- during ripening time.01) than those of goat’s (460 mg 100 g 1 DM) or cow's D2 (19. goat’s and cow’s milk.3 not differ (P > 0. E. 7. The kind of milk overall statistical analysis showed that.. Tau (which origi- D2 (35.14%) sheep’s (380 mg 100 g 1 DM) milk cheese. 1995). 7. Effect of type of milk matrix and could have increased the rate of proteolysis (Chapot- Chartier. Centroids as well as in ewe’s and cow’s milk cheeses (<1% of TFAA.001).05). 8).7 cheese (P < 0. Linear discriminant functions (D1 and D2) for cheese samples at 1. regardless of the kind of milk and culture used). In contrast.86%). Buriana and Farag -1.1 mg 100 g 1 DM) milk cheese. Mallatou et al. thermophilus Fig. in 180 day 0. Total free amino acid mean content in cheeses made with sheep’s. Linear discriminant functions (D1 and D2) for cheese samples made from mean concentration of Orn in cheeses made from ewe’s milk was sheep’s. lactis and shows higher specific the mean TFAA concentration from 263 mg 100 g 1 DM in 1 day old cheese to 590 mg 100 g 1 DM in 180 day old cheese.001). the overall statistical analysis showed that the mean 200 goat's milk content of TFAA in Teleme cheese made with the thermophilic cul- ture was 551 mg 100 g 1 DM and was significantly higher than cow's milk 0 that in mixed (421 mg 100 g 1 DM) or mesophilic (350 mg 100 g 1 DM) culture (P < 0. (1998) found that the TFFA content in Picante cheese made from sheep’s milk was lower than the cheese made from goat’s milk too. the TFAA in cow’s milk sheep's cheese had a mean content 481 mg 100 g 1 DM and was significant goat's 2.2 mg 100 g 1 DM and was significant lower (P < 0. Low Tau contents have also been reported for Picante cheese (Freitas -1. the cheeses were cor- 180 days of ripening as well as the day cheeses entered the cold room (R). D1 (54.81%) while TFAA and Asn contributed mainly to discriminant function Fig. there was an increase in peptidase PepS) with respect to Lc.0 mg 100 g 1 DM and was signif- 1 icantly higher (P < 0. 60 According to Mehaia and Al-Kanhal (1992).9 120 nates from Cys) is the most abundant FAA in goat’s milk. Moreover. Cheese made with Centroids goat’s milk was also significant higher than that of sheep’s milk 0. The highest TFAA contents of Teleme cheeses made with the Fig. K.14%).7 vine cheese from 297 mg 100 g 1 DM to 784 mg 100 g 1 DM.9 old cheese. but accumulated as a metabolic product of microorganisms. AAB and GABA) did not originate from case- in. 1996). 6. thermophilic culture.05). 5.3 (1983) found that the TFFA in ovine Teleme cheese was higher than that of bovine cheese. . from 273 mg 100 g 1 DM to 619 mg 100 g 1 DM and in 400 cheese made with the mesophilic culture from 246 mg 100 g 1 DM to 530 mg 100 g 1 DM.C.1 et al.9 R other two types of milk.4. Stepwise discriminant analysis selected 18 amino acids for dis- criminating among the three kinds of milk (Fig. cases). Cheeses made with the mesophilic 0 60 120 180 culture had significantly lower (P < 0. -7 -4 -1 2 5 8 Orn has been found in Teleme cheeses from the first day (Table D1 (80.7 higher (P < 0. it is known that S. Effect of type of culture Total amino acids mg/100g DM 1000 The mean concentration of TFAA in Teleme cheese made with 800 the thermophilic culture increased from 366 mg 100 g 1 DM in 1day old cheese to 877 mg 100 g 1 DM in 180 day old cheese. Sotirakoglou / Food Chemistry 111 (2008) 606–615 613 Plot of Discriminant Functions 8.001) mean concentration of Ripening day TFAA than had cheeses made with the other two types of culture.001) than that of cow’s (13.05). In this 180 study.05). Freitas et al. TFAA and -7 -5 -3 -1 1 3 5 Phe contributed the most to discrininant function D1 (80. In ovine cheese. sheep's milk Generally. Two discrimi- -3. The Fig. in caprine Plot of Discriminant Functions cheese from 326 mg 100 g 1 DM to 652 mg 100 g 1 DM and in bo- 4. 60. could be due to an earlier lysis of thermophilic starter bacteria that could have enhanced the early release of intracellular enzymes in cheese 3.44%) 2. Pappa. Both D1 rectly classified according to the kind of milk (100% of the total and D2 were significant (P < 0. 3. 120. As it can be observed in Fig. in 600 Teleme cheese made with the mixed (thermophilic. D2 (19. Tau was found in low concentrations in goat’s milk cheeses. on the corresponding days (Fig.3.9 Ripening time cheese had a mean content of 16.001) than that of the cheeses made from the 4. (2004) found that the TFAA contents of Teleme cheeses made from ewe’s or goat’s milk did -3.mesophilic) culture. regardless of the type of milk used. 7). Orn in goat’s milk 6. goat’s and cow’s milk. 1998) and Idiazabal cheese (Barcina et al.1 nating functions were statistically significant (P < 0.86%) 2).

E. Two dis. Cheeses Polo. 70.. K. M. P. 6. Prietro. M.. Linear discriminant functions (D1 and D2) for cheese samples made using a IDF (1986). Protection. Journal of Agricultural and Food Chemistry. USA: Waters Association. F. C... culture. lar results when they manufactured Hispanico cheese with MA. 16.614 E. H. & Nunez. Goat cheeses ranged over intermediate levels. Tzanetakis. Barcina. culture.9 Proteolysis in Turkish white-brined cheese made with defined strains of -6 -4 -2 0 2 4 6 Lactococcus. The variation of amino acids in Telemea cheese 5.. Upadhyay. 9). (1981). 179–206. Pico Tag Amino Acids Analysis System. Medina. 17. Journal of Dairy Science. Fig. G. Ramos. L. & Marth. N. E. Bosman. PepN and PepC (Rul & Monnet. J. H. (FIL- thermophilic (T). Lait. Kandarakis. Xavier Malcata. M. Thermophilic culture 200 Moreover. A. (1990). TFAA and Leu contributed the most to discrininant function D1 570–581. H. P. Evolution of free amino acids during Idiazabal cheese ripening. Proteolysis in cheese during ripening. C.. (2004).. L. P. Hannon. L.54%). M. M. A. Greek Codex Alimentarius (1998). P. C. Egyptian Journal of Dairy Science.. P. In National Dairy culture. & Sanchez. 351–353. the correct group by the type of culture was 88. thermophilic or mesophilic adjunct culture. although some liquid chromatography and peptides by gel electrophoresis in Mahon cheese overlapping between mixed and mesophilic cultures was during ripening. 50. 14. Athens.. M.. 76. and Nunez (2002) found simi. International Dairy Journal. G. 9. Taurine and other free amino acids in milk higher aminopeptidase activity than did mesophilic adjunct cul- of camel. (2004).. Athens. 187–194. A. Zerfiridis. M. 273–290. (1993). E. E. Both D1 and D2 were significant (P < 0. Lactobacilli-their enzymes and role in ripening and spoilage of cheese: A review. 53–59. 899 article 83 paragraph D3C.. biochemical and sensory changes in Turkish white-brined cheese during ripening. & Coolbear. criminating among the three types of culture (Fig. 4). Cheese and processed cheese products. 599–610. & Vafopoulou. Influence of milk source and ripening time on free amino acid profile of 1. Fox. J. C.. goat.. Pappa. Determination of dry matter. (2002). Amsterdam: Elsevier Academic Press. (1996). Journal of Dairy Science. P. (1992). 977–987. and Nunez (2001) concluded that thermophilic ad. Teleme cheese from deep-frozen curd. C..1 using a Lactococcus lactis ssp. Medina.. M. & Nunez. & Boumba. A. (1991). Milchwissenschaft. M. function D2 (5. & Torre. (1979). 27–42). 12. Total free amino acid mean content in cheeses made with the thermophilic Anifantakis. Les autolysins des bactéries lactiques. M. Anifantakis. H. 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A. a mesophilic (M) or a mixture of thermophilic–mesophilic (MX) IDF standard no.. E.. Polychroniadou. the kind of culture added and the ripening time. 14. I. Magboul. P.54%) Centroids Freitas. I. overall proteolytic activity of lactobacilli has been found to be Mallatou. A. F. 88. E. A. H. J. D2 (5. Garde. (2006a).46%) Hayaloglou. R. & Ordonez. L. 31. P.46%). Pappa. culture Chapot-Chartier. FEMS Microbiology Reviews. activities of PepX. 9. levels (Khalid & Marth. Y. The total free amino acid (TFAA) content increased 600 with ripening time. Garde. a thermophilic starter and bacteriocin-producing Lactococcus lactis subsp.05). Fresno. I. cheeses made with cow’s milk had higher TFAA contents Thermophilic-Mesophilic culture than had cheeses made with ewe’s milk. 81–86.

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