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Food Chemistry 111 (2008) 606–615

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Changes of free amino acid content of Teleme cheese made
with different types of milk and culture
Eleni C. Pappa a,*, Kyriaki Sotirakoglou b
a
National Agricultural Research Foundation, Dairy Research Institute, 452 21 Katsikas, Ioannina, Greece
b
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-
Keywords:
philic culture. Cheeses made with the mixture of thermophilic–mesophilic culture ranged over interme-
Teleme
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.
Milk
Ripening

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: instgala@otenet.gr (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.
doi:10.1016/j.foodchem.2008.04.027

Eluent B was 45% acetonitrile. goat and cow milk. This was repeated five times. sonicated in a water bath for 20 min and centrifuged at Laboratorium. S. Separations were conducted at 45 °C and the absorbance was mon- The experimental design was that described elsewhere (Pappa. Cohen. mixed or mesophilic culture. culture (M). Each kind of milk was used in making taining 0. Lb. goat’s or cow’s tives by RP-HPLC using the PICO-Tag amino acid analysis system milk. a mesophilic or a mixture of a thermophilic.5 min using curve 8. 500 ll of supernatant were diluted (1:1. an ice bath and then centrifuged at 20.55 with acetic acid con- After manufacture. v/v) coccus thermophilus and Lactobacillus delbrueckii subsp. injection. Milford. until both pH and moisture values reached the limits of and 15% methanol. thermophilic–meso. These results could serve as a model for fu. near the Dairy Research Institute where the as follows: 1 g of cheese was weighed and 10 ml of 0. Samples were suspended in 100 ll of Pico Tag sample milk was used in making the three variables of cultures and nine diluent (Waters. The objective of this work was to study the effect of three com. Horsholm. France). USA).000g for 10 min. Cheese-making and sampling ing retention times with pure standards. Materials and methods nium 32 software package. then 3–6% eluent B made from sheep’s milk using thermophilic. Eluent A comprised at 19 °C. and Tarvin (1984). bulgaricus. Run time was 67 min with flow rate 1 ml min 1. K. To our knowledge. E. Denmark) was added at a The solvents were of HPLC grade. transferred to the cold room at 22. lactis. 13. Experimental design volume of 20 ll of the amino acids was injected into the column. FRC-60 (Hansen’s Laboratorium. respectively and afterwards cheeses were trans. Pappa. A Waters reverse phase column (3. delbrueckii subsp. (Waters. Before each injection.0 g per 100 kg milk. holding 24. respectively. the column was equili- 64. except that glutamine (2. MO.5% acetonitrile. on free amino The FAA were identified as phenylthiocarbamyl (PTC)-deriva- acid release of Teleme cheese made from sheep’s.1. itored at 254 nm. Gradient elution was as follows: ferred to a cold room (5 °C) for up to 6 months. thermophilus. standard. R-703 (Hansen’s Laboratorium. 60. and curve 11 maintains starting conditions days. Curves 5. Analysis of free amino acids (FAA) and chemical analysis mercial concentrated starters. lactis and Lactococcus bated for 10 min at room temperature and then they were vac- lactis subsp. To assist curdling (St. 20 ll of derivatizing solution ganismds was 1:1) and (iii) a freeze-dried concentrated mesophilic (methanol: water: triethylamine: phenylisothiocyanate 7:1:1:1. was manufactured using (a) sheep’s milk of the ‘‘Boutsiko” breed The method of Bidlingmeyer. to obtain a first quality cheese that is 656% moisture v/v/v). at a quantity of 0.. with 40% trichloroacetic acid. 2006a). Methionine sulfone was added as an internal standard and P43% fat in dry matter (Greek Codex Alimentarius. The samples were incu- the microorganisms Lactococcus lactis subsp. Denmark). respectively. cremoris. consisting of the microorganisms Strepto. there Cheeses were sampled for analysis on days 1. Drainage was under ical grade. HPLC consisting of a controller (model 600). methionine sulfone and glutamine were purchased from Sigma tioned was added to it. linear and concave gra- ophilic culture were transferred to the cold room at 23.64 and 0. a solvent pump (mod- el 600E). standardized milk was pasteurized at 63 °C for 30 min and then Standard mixtures of basic. acidic and neutral amino acids and cooled at 37 °C.75. (the ratio of thermophilic to mesophilic microor. Curves 5.90  30 cm) was used. 1998). respectively and 6. Sotirakoglou / Food Chemistry 111 (2008) 606–615 607 with the same type of milk and culture. Tag system. 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. (Hansen’s Laboratorium. Teleme cheeses 0–3% eluent B over 13. IL. cheeses were ripened in the warm room taining 2. 8 and 11 are gradients programmed by the Waters HPLC con- cheeses made from cow’s milk using thermophilic. then 6–9% eluent B over 6 min using philic or mesophilic culture were transferred to the cold room at curve 5.4 mM methionine sulfone. Kandarakis. mixed or mes.5% (v/v). bulgaricus and 1 M sodium acetate 1:2:2. at 33 °C. one of the cultures already men. which were vacuum-dried again. detail in the PICO Tag amino acid analysis Operator’s Manual pa et al.6 and 656%. Louis. cremoris. FYS-11 (Rhone-Poulenc blender. Samples were applied using a rheodyne injector (model 7725i). 20 ll of a redrying solution (triethylamine: methanol: Lactococcus lactis subsp. v/v/v) were added to the dried samples. respectively. The working amino acid standard was prepared as described in Teleme cheese was manufactured as described previously (Pap. respectively. cheeses made from at 34% eluent B for 12 min. 2. Anifantakis.1 M HCl. . USA). 70 mM sodium acetate adjusted to pH 6. 2006a).45 lm pore size before day after day basis. Samples were analyzed on a WATERS ture research in this field.3.C. a helium degasser and a tunable absorbance detector (model 486).73 for the sheep. After cooling. until the next step. The (standard solution: internal standard 1:1. namely a thermophilic. 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. Phenylisothiocyanate (PITC) (sequential of milk CaCl2 solution (40%. Rennet powder HA-LA grade) and thiethylamine were from Pierce (Rockford. during ripening. w/v) was added. each kind of uum-dried. USA) and were filtered through a lots of cheese were obtained. mine: acidic and neutral amino acids: basic amino acids 1:1:1. 25 ll of philic culture (MX). Free amino acids were extracted from cheese Friesian cows. That is. On the day of cheese-making. on a HV hydrophilic filter (Millipore) with 0. then followed by a purge at 100% elu- goat’s milk.5 min and re-equlibration to 100% eluent A. The solvent system consisted of two eluents made pressure and salting took place in brine (14% NaCl. USA). w/w) for 16 h according to the Operator’s Manual (1984). con- cheese-makings took place.mesophilic culture. 6 and 8 are convex. The mixture was homogenized using an Ultra-Turrax dried yogurt thermophilic culture (T).2. All other reagents were analyt- quantity of 3. MA. were ent B for 4. was added as the internal Teleme cheese with three starter culture variables: (i) a freeze. whereas. using thermophilic. then 9–34% eluent B over 20 min using curve 6. Teleme cheese HPLC analyses were done in duplicate. brated with solvent A for 20 min. Identification of the cheese amino acids was achieved by compar- 2. the deproteinized supernatant were vacuum-dried in the PICO- consisting of the microorganisms Lactococcus lactis subsp. Briefly. too (gluta- ratios of 0. 16 and 12 days. profile of Teleme cheese. equipped with a 50 ll sample loop. The mixture was left for 10 min in (ii) a freeze-dried concentrated mixture of thermophilic–meso.5 mM) was added. v/v). Denmark) consisting of v/v/v/v) were added to the dried samples. 11 days. 17 and 15 dients. The chromatograms were processed using the Millen- 2. trol station. 40% water (19 °C). Milford. & Zerfiridis. MA.5 min using curve 11. A 2. 3000g for 10 min. 0. 120 and 180. over 10. the milk was standardized to casein to fat (1984).

while -6 -4 -2 0 2 4 6 cheeses 120 to 180 days old were placed on the negative side of Component 2 (10. Rock- acids. Leu Arg Lys Ile Val Orn indicating that they were positively correlated with each other. Ser B_Ala Component 3 (10. goat’s or cow’s) as factors and ripening time as Asn Leu -2 split–splot factor. together with the intact FAA. a comparison of the values of each Cit Met parameter of Teleme cheese manufactured with one type of milk -4 Gaba (sheep’s. USA and GraphPad Prism version 4.608 E.75% of the total variability. Cheeses made from goat’s milk were clus- tered on the negative side of PC3. Plot of the free amino acids as a function of the two first principal components. near Pro and Ala and therefore 3. 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. placed on the positive side of PC3. On the other hand. In Fig. a number of pal component could be a representative of the kind of milk. 1986). TFAA AspThrHis Tau AlaCys Lys Ser plied to the data. Chicago. Results and discussion and Ser (positive value). Cheeses made with thermophilic culture were clustered near (SPSS for Windows version 14. K.68% of the total variability and was defined by AAB. Linear regression analyses placed close together on the positive side of PC2. However. Principal component analysis. because all cheeses. standard solutions of pure amino acids had eigenvalues greater than 2. these amino acids were ly- time. a common statistical cut-off point. 2). PCA of the 27 free amino acids resulted in eight principal components with eigen. goat’s or cow’s) with the three types of culture (thermo- philic. free amino acids were represented as a function Cit of both first and second principal components.68%) PC1 (Fig.75%) with one type of milk and one type of culture. than cheeses made with the other cultures. sented the opposite characteristics.14% of the total vari. 1. San Diego.05). B_Ala AAB 4 Component 2 (10. These amino acids were Gln Phe His placed closed together on the negative side of the horizontal axis. at different ages. Fig. 1 are ripened 180 d old cheeses. indicating a high were applied to fit the evolution of free amino acids with ripening positive correlation.4. the third princi- During the extraction and derivatization process. -2 Tau They were away from the axis origin. having the highest contents of TFAA.8.0. Principal component analysis. Gly and Lys. 0 GlyPhe Orn mixed thermophilic–mesophilic or mesophilic) and the kind of Gln Arg Tyr milk (sheep’s. 2. The third principal component explained another 10. Statistical analysis Val Ile Glu Trp 2 Pro A three-way split–splot analysis of variance (ANOVA) was ap. Plot of the free amino acids as a function of Asn.2. In Fig. considering the type of culture (thermophilic. were co-injected with the standard mixture solution to identify peaks in the standard solution.42% of the total variability and was defined by Pro. Moreover. IL. Consequently the second prin- USA). All the cheeses that were located on the left of 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. STAT- AAB and b-Ala and therefore had higher contents of these amino GRAPHICS Plus for Windows version 5. were clustered on the positive side of PC1. USA. CA. 4 values greater than 1. The least significant differences were obtained using the LSD test Fig. suggesting that they were well represented from the first PC.85% of the total variability and fied peaks. Asn. Ala (negative values) 3. The eight selected components accounted for 79. . The second principal component explained the second and third principal components. generally. MD. 2 AAB and b-Ala were for selecting discriminant variables.. ing opposite to Gaba. cipal component could be a representative of the type of culture. The unidentified peaks were present. since a great correlation be- Biplot tween the free amino acid contents was noticed.1. these cheeses had the lowest content of Ser. On the other hand. 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. from 1 -4 to 60 days old. these peaks did not interfere with the identi- together they accounted for 51.C. and the results were ex- 6 pressed on a dry matter basis. 1. General had the highest contents of Pro and Ala (Fig.42%) 2 Asp AAB ability. Manugistics Inc. at each -6 age. 1). therefore they were negatively correlated to Statistical analysis was performed using statistical packages this. which could be considered as Ala Pro representative of the ripening time. (P < 0. Consequently. When necessary. and was Met Asn Gly Trp Glu 0 TFAA mainly defined by TFAA. SPSS Inc. 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. was made. Gly and Lys. first three principal components were the most important. b-Ala (posi- milk and the stage of ripening.0. mixed thermophilic–mesophilic or mesophilic). which repre- ville. The first principal Cys Thr Gaba Tyr component explained 30.68%) 2. Pappa. 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. Wilk’s lambda (k) criterion was used tive values) and Gaba (negative value).. the type of 10.

985 *** 0.973 *** 0. Barcina.05) and Tau.993 *** 0.736 *** 0.937 *** 0.988 *** 0.913 *** 0.994 *** 0. Effect size Sig.8 in most cases). kind of milk. 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. Effect size Sig.934 ** 0. NS: not significant.908 *** 0.823 *** 0. Zerfiridis.985 *** 0.960 *** 0.993 *** 0.934 Leu *** 0.939 *** 0. in Cheddar cheese 1592 mg 100 g 1 DM.970 Val *** 0. K.868 *** 0.911 *** 0. 1995).986 *** 0.939 *** 0. 4 it can be seen that Leu.900 Pro *** 0. 0 60 120 180 vidual FAA of Teleme cheese made with different starter cultures Ripening day and milks during ripening.984 *** 0. Franco.971 *** 0. as was expected since.806 *** 0.979 *** 0. The same FAA were found in abundance cold room. All two-way and three- way interactions between the factors were significant (P < 0.901 *** 0. which was not significantly different with respect to starter addition (P > 0. At the age of 180 nificantly from 295 mg 100 g 1 DM in 1 day old cheese to days.903 *** 0.883 *** 0. TFFA mean content of Teleme cheese increased sig. **P < 0.811 *** 0. weeney.05) 200 for the TFAA and for all individual free amino acids.984 *** 0.940 *** 0.944 *** 0. 2003).979 *** 0. type of culture (C) and ripening time (T) Amino acids M C T MC MT CT MCT Sig.949 Met *** 0.590 *** 0. Val and after 180 days of ripening and re-increased rapidly until day 360.990 *** 0. (2004) and Polychroniadou and 492 mg 100 g 1 DM in 120 day old cheese and finally to 675 mg Vlachos (1979). Buriana and Farag (1983) found that the TFFA decreased From Table 2 and Fig. these compounds are released by proteolytic agents.740 Arg *** 0.984 Phe *** 0. Ibanez. type of culture and rip.921 Gaba *** 0.953 *** 0.966 *** 0.966 *** 0. Fig.980 Ile *** 0. were significant (P < 0.907 *** 0. & Xavier Mal- study.962 *** 0.598 *** 0. Effect size AAB *** 0.928 *** 0.996 *** 0.994 *** 0. centrations of almost all FAA showed a clear tendency to increase with ripening time.983 *** 0. regardless of the type of milk and culture used.509 *** 0. Guven.986 *** 0. increase of hydrophilic peptides and decrease of hydrophobic Edam cheese 1056 mg 100 g 1 DM and in Romano cheese peptides.896 *** 0.824 Cit *** 0. 2001.984 *** 0.839 His *** 0.875 *** 0.796 *** 0.988 *** 0.990 * 0.634 *** 0. In Table 2 it can be seen that the con- Fig.986 *** 0. Hannon. 2005. they accounted for 41. Table 2 shows the concentrations of indi.861 * 0.902 *** 0. 3 presents the evolution of mean total free amino acids (TFAA) of Teleme cheese during ripening. decrease of the intact casein con. & McSweeney.905 *** 0.930 Orn *** 0. Ibanez.913 Glu *** 0.001). Effect size Sig. Phe. which describes the pro- 0 portion of total variability attributable to a factor.961 *** 0. except Ala.985 *** 0.05.926 *** 0.993 *** 0.934 *** 0. & McS- found in this study. is in accordance with the results found by Mallatou et al.5% of the TFFA.970 *** 0.954 Asp *** 0. E.997 *** 0.897 *** 0.894 *** 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.990 *** 0. Fox.964 *** 0.980 *** 0.903 *** 0. The FAA in Manchego cheese typically contribute the increase of nitrogen faction. Vicente.668 ** 0.880 *** 0. in tent.983 *** 0. 400 ening time (P > 0.990 *** 0.949 *** 0. Pappa.01.914 *** P < 0.980 *** 0. during proteolysis.951 *** 0. The increase of TFFA at all stages of ripening.965 *** 0.952 *** 0.994 *** 0. which has a high content Table 1 Significance levels and size effect of the three-way split–splot ANOVA for the effects.984 Tyr *** 0.742 * 0.940 *** 0.936 *** 0.962 Tau *** 0. Hayaloglou.434 *** 0. The levels of TFFA Fox.939 *** 0.824 *** 0. regardless of the type 316 mg 100 g 1 DM on the day cheeses were transferred to the of milk and culture used.973 TFFA *** 0.979 *** 0.001) for the total free amino acids (TFAA) of Teleme cheese and for all of the individual free ami- no acids.971 *** 0.948 *** 0. The proteolysis of as1-casein. Effect size Sig.872 *** 0.930 Ser *** 0.946 *** 0. as well as in other cheeses (Hayaloglou.852 *** 0.860 *** 0. Effect size Sig.960 *** 0.908 *** 0.078 *** 0.2.. kind of milk (M).829 *** 0. Barcina ies.982 *** 0.C. are within the range reported from other stud.974 *** 0. Glu.996 *** 0.423 *** 0.965 *** 0.971 *** 0.991 *** 0. cata.991 Cys *** 0.952 Bala *** 0. 3.915 *** 0.900 Thr *** 0. 100 g 1 DM in180 day old cheese (P < 0.923 *** 0. which did not change significantly during rip. Generally. Effect size Sig.975 *** 0. .959 *** 0.974 ** 0.931 Gln *** 0.993 *** 0.978 *** 0.882 *** 0. Sotirakoglou / Food Chemistry 111 (2008) 606–615 609 3. Kand- arakis.912 *** 0.867 *** 0.909 *** 0.942 *** 0. Anifantakis.577 *** 0.995 *** 0.964 *** 0. 1728 mg 100 g 1 DM.990 *** 0.967 Lys *** 0.869 *** 0.05).925 *** 0.970 *** 0.958 *** 0.493 *** 0.980 *** 0.989 *** 0.732 *** 0. to 424 mg 100 g 1 DM in 60 day old cheese.932 *** 0.981 0.982 *** 0.977 *** 0.938 Ala *** 0.849 *** 0.989 *** 0.988 *** 0. to in Teleme cheese by Mallatou et al. (2004). & Barron. Javier Carballo.953 Gly *** 0.958 *** 0. Barcina. 2004.833 Asn *** 0.550 *** 0.989 *** 0.938 *** 0.938 *** 0. and Ordonez (1995) reported an increase from et al.926 *** 0. *P < 0.964 *** 0.683 *** 0.863 *** 0. Table 1 also shows the effect size (>0. in this 650 mg 100 g 1 DM (Tavaria.943 *** 0.993 *** 0. Lys were the major FAA of Teleme cheese at all stages of ripening. as estimated by bal cheese.996 *** 0.912 *** 0.957 *** 0.994 *** 0.866 *** 0. Evolution of total free amino acids during the ripening time.001.620 *** 0. Pappa.966 *** 0.886 *** 0.966 *** 0. 600 ening time. Guven.940 *** 0.985 *** 0.910 NS 0.988 *** 0.990 *** 0.925 *** 0.991 *** 0.916 Trp *** 0.965 NS 0.

91b 7.42a 10.91b 1.81a 3.51b 19.41b 12.02a 4.12b 12.51a 9.11a 3.13a 19.11a 2.12a 32.71a 5.21a 0.22a 5.62a 40.91a 2.62a 2.01c 3.02a 11.22a 8.412a 10.02a 4.41a 1.01b 0.82c 0.51b 3.63b 15.91b 14.11a 18.62b 0.01b 14.22a 2.11b 24.91a 2.81b 4.11b 4.112c 19.41a 2.12ab 0.72b 2.11b 22.31a 8.41b 3.21c 3.73b 17.01a 0.22a 0.71b Leu 23.01a 3.02a 1.31a 0.02a 20.912a 23.21b 1.52a 9.22a 1.31a 3.11b 2.31b 5.32b 17.81a 1.31a 3.62a 35.05) between kinds of cultures.02a 0. K.31c 37.01b Ile 40.02b 2.91b 4.12b 1–3 : For each age and each kind of milk.81a 8.12a 3. B R: Day cheeses were transferred to cold room.42b 7.33a 2.71b 5.32b Cit 0.81a 25.01b 28.21c 6.12a 1.71a 10.81c 5.21a 8. .42a 4.92a 2.52a 11.72a 34.82a 17.02a 15.71a 5.12b Gly 6.42a 5.71a 11.11b 17.412b 4.71ab 13.02a 11.91b 11.51a 0.11b 16.53b 55. a mesophilic (M) or a mixture of thermophilic and mesophilic (MX) starter during ripening Age.22a 63.53a 34.13c 67.51a 2.41a 16.12a 6.81c 8.02b 0.22a 9.42a 20.22a 0.21b 3.312c 11.12b 4.93b Val 16.81a 0.33a 6.62a 2.31a 6.52b 3.33a 19.C.52a 3.82a 9.72b 3.23a 51.91a 6.82a 5.82a 12.312b 15.22a 1.12b 11.03a 20.81a 9.72b 1.41b 7.52a 35.92a 8.72a 0.52b 7.42b 11.71a 33.91b 14.61a 1.91a 3.21a 37.41b 2.12a 12.31c 2.71a 1.11b 19.42a 31.01b 16.01a 17.72a 0.81a 0.51a 13.91b 3.02a 6.52b 21.02b 0.02b 2.01c 4.43c Gaba 3.72a 38.81a 17.22a 10.63a 17.82b AAB 26.71b 0.33b 0.82a 5.52a 46.21a 3.82a 6.01b 38. Pappa.93a 21. Sotirakoglou / Food Chemistry 111 (2008) 606–615 Glu 30.73a 43.92a 44.12a 5.32a 16.42a 21.81b 19.61b 6.012a 1.52a 0.12a 26.42b 4.02a 2.42a 3.43b 2.02a 0.01a 2.05) between kinds of milk.52a 32.72a 35.01a 6.81ab 2.43c Ser 3.82b Thr 5.01a 4.41a 21.91b 16.63c 18.71b Lys 66.11a 29.61a 13.71a 6.31a 49.112a 3.51b 16.42a 3.31a 6.712a 0.62ab 2.91b Asn 12. different numbers indicate significant differences (P < 0.93b 8.92a 3.11a 2.11b 6.81a 6.01c 37.82b 8.82a 0.71a 6.21b 5.01a 1.81a 0.71a 0.81b 1.92b 18.01a His 4.21b 13.03b 84.21a 13.93a 19.02a 0.61a 1.61a 3.91a 73.92a 16.11b 0.81b 7.12a 10.93a 9.41b 13.31ab 1.52a 3.61ab 9.52a 8.61b 3.33a 23.61a 6.51c Tyr 11.21b 4.42ab 7.13a 29.92b 5.61b 2.51a 15.02b 1. A DM: Dry Matter.51b 8.512a 26.01a 8.81b 7.31a 1.61a 1.72a 4.81a 4.71b 28.62a 29.12a 4.72b 13.81b 26.22a 0.62a 0.01a 1.71c 8.12a 11.412b 25.22a 3.81a 9.11a 11.71a 1.12c 5.91a 13.52a 7.02a 9.11a 1.33b 56.01a 6.43c Cys 6.212b 14.71b Met 18.52b 33.61b 5.71a 13.01b 5.52a 64.92a 7.71a 3.92a 13.02b 58.92a 1.81a 9.82b 3.02b 2.92a 16.312a 7.91a 0.92a 15.21a 10.91a 0.03b 0.01a 0.72c 36.83a 3.212a 32.52b Trp 2.23a 7.21a 2.81b 17.22b Gln 7.61a 4.92a b-Ala 16.62a 7. (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 1.01b 5.81a 8.81ab 19.32a Arg 27.01b 6.12a 7.212a 3.72a 6.61a 1.32b 22.21a 1.52a 3.81b 6.21a 6.92b 33.51a 12.81a 0.32a 16.42a 0.01a 0.71a 14.21b 5.11a 35.52a 2.02a 9.71a E.21a 14.21a 3.72a 0.81b 24.812ab 12.22a 3.81a Orn 14.41a 52.82a 7.21b 9.91b 2.81b 15.12ab 5.81c 2.32a 34.93b 0.93a 28.01a 8.81a 4.12c Tau 2.81b 68. different letters indicate significant differences (P < 0.21b Pro 7. 610 Table 2a 1 Changes of amino acid contents (mg 100 g DMA) of Teleme cheese made from sheep’s (S).92a 7.42a 8.22a 15.21a 3.51ab 6.42a 0.41a 4.41c 22.91a 16.02b 0.52a 15.81b 12.21b 11.22a 84. %.11b 31.11b 1.11 2. goat’s (G) or cow’s (C) milk using a thermophilic (T).12c 8.11a 11.21a 1.32b 7.71a 0.33c 0.61b 7.62a 6.12a 23.11b 1.51b 24.21b 3.21b 7.02a 36.32b 8.12a 1.02ab 16.02c 4.712a 2.31b 23.41a 25.11b 2.71a 3.12a 1.61a 11.01a 2.92a 26.53c Phe 22.02a 1.31b 5.52a 1.92c 1.71b 23.41a 1.31a 1. a–c : For each age and each kind of culture.32b 23.51a 2.31b 16.41b 23.71a 10.52a 0.72c 4.32a 11.21b 7.02ab 13.82c 2.51a 8.21a 13.712a 8.83a 22.02a 10.81a 6.11a 6.91a 5.93b 1.11b 38.81b 12.91a 0.41a 1.22a 11.42a 1.22a 11.32b Ala 3.81a 43.21a 29.61a 14.02a 8.61a 14.91b 1.31b 10.52b 3.81b 3.81b 1.11b 10.81b 5.02b 7.31b 3.

11a 30.81a 1.41a AAB 46.52a 28.41c 41.43b Glu 122.81a 2.11a 21.32a 1.71a 19.41b 11.42ab 0. A DM: Dry Matter.31a 49.51b 10.41b 9.61a 6.5a Arg 29. 611 .41a 31.12b 2.22a 1.51b 1.82c Val 28.01a 4.41a 28.21b 9.22b 23.53b 3.91b 14.51a 13.91a 2.61a 18.22a 20.91c 15.11a 9.11b 4.12a 13.83c 66. K.93a 140.73a 43.02a 5.22a 4.31a 3.91a 7.62b 0.12a 12.91a 12.81b 19.03a 41.12a 12.42a 38.91a 4.61a 15.03a 0.71a 28.05) between kinds of milk.63ab 9.512a 10.62b 1.11a 2.01a 11.31a 6.03a 43.112a 15.82a 1.01a 3.21a 8.01a 2.02c 10. a–c : For each age and each kind of culture.12b 0.73a 31.52b 4.81a 0.31a 9.11a 2.21a 10.61b 0.23c 70.73b 4.71c 7.01a 28.12b 34.21ab 13.91a 48.02ab 10.91b 10.31a 2.41a 5.62c Met 20.01a 38.11a 12.51a 44.21b 14.92b 5.81a 3.02a 71.21a 3.71a 3.42a 42.01a 23.62a 11.42b Cit 3.51a 1.11a 11.51a 9.51a 11.52b 4.72ab 23.32a 7.41a 26.31a 13.31a 8.72a 5.12b 8.91b 17.91b 1.91b 16. %.42a 12.42b 13.41a 0.62a 3.51a 2.92a 6.61a 16.31a 4.12b 11.71a 4.12a 1.22a 4.42a 19.52a 2.212a 26.63a 28.12b 19.01a 0.41a 19.12a 12. goat’s (G) or cow’s (C) milk using a thermophilic (T).21a 15.82a Tau 3.72b 30.22a 6.41b 10.91b 30.31a 38.61a 2.73b 48.62a 0.11a 20.41a 5.81b Trp 7.21a 29.42b 58.91b 5.11c 12.71c 38.81a 4.72a 72.61b 3.31a 13.61a 30.22a 3.01b 7.41b 2.81c 40.42a 9. Pappa.32a 3.72c 11.42c 10.312a 16.11a 3.53b Cys 6.51a His 13.92b 40.01a 52.91a 4.91b 6.73ab 4.22c 61.52b 12.02b 0.31b 13.81b 17.72c 3.51b 0.81b Pro 8.Table 2b 1 Changes of amino acids content (mg 100 g DMA) of Teleme cheese made from sheep’s (S).02a 5.43b 18.11b 33.71b 18.12a 0.11b 1.41a 40.71b Ala 4.912a Tyr 6.02b 22.51b 1.21c 24.11a 6.52a 36.91a 3.93a 1–3 : For each age and each kind of milk.42b 16.91b 3.72b 21.71b 46.61b 8.82b 20.21a 24.81a 7.42a 6.01b 5.51a 11.22a 6.61c b-Ala 23.31a 0.92a 32.51b 77.11b 7.12c 7.83a 22.01a 27.412a 132.71a 8.51b 34.61a 0.52a 8.82a 37.03b 9.31a 4.52a 27.01b 5.12b 6.33b 5.012a 22.72a 14.02a 13.21a 18.63b 22.52a 11.21b 33.41a 23.11b 0.21a 20.41a 4.32a 2.31a 4.C.52a 8.41b 34.22b 12.01ab 3.62a 6.31a 4.01b 25.42a 1.03c Lys 59.53b Ile 35.41a Asn 5.61a 8.82b 6.71a 4.92a 12.92a 20.52a 6.51b 15.32b 8.11a 1.82c 6.51a 5.62b 23.41a 13.52a 38.81a 1.412b 25.81a 3.91a 0.33b Orn 7.62c 59.12b 3.41a 16.62c 2.02a 0.02a 3.41a 0.21b 2.51a 5.53a 35.01c 147.41b 6.01b 77.93b 17.82a 6.312a 14.11a 4.61b 26.91b 157.23b Gaba 3.71a 38.61b 15.01a 3.81a 5.91b 17.32b 16.62a 13.51c 28.02a 5.02b 1.05) between kinds of cultures.02b 52.012a 64.512a 7.41b 3.52a 30.01b 2.42b 3.83b 8.23b 4.51b 15.01a 2.11c Leu 16.62a 13.92a 21.11a Thr 15.71a 0.01ab 3.12b 12.61a 6.72a 12.21a 3.91b 20.91a 0.31b 14.42a 18.11a 5.41a 3.51a 1.22a 3.11b 3.31b 27.92a 9.12a 134.92b 16.01a 6.81b Gly 5.62c 9.03a 9.61a 7.22b Phe 67.92b 3.71a 6.21a 4.91b 4.93a 37.72b 6.11b 135.41a 8.81b 5.72a 2.412a 2.92a 3.02c 114.23c 46.912a 1.812b 3.91b 11.11a 81.82b 26.82b 2.91b 5.82a 18.92a 3. a mesophilic (M) or a mixture of thermophilic and mesophilic (MX) starter during ripening Age.11a 1.91a 4.31a 11.92ab 0.81a 2.01c 84.71a 5.71a 51.01a 78.02b 9.01b 9.01a 1.62b 9.52a 26.61c 2.81b 36.01b 101.62a 0.01a 30.81ab 8.42a 36.21a 3.62b 51.41b 13.13b 3.82a 7.11a 1. Sotirakoglou / Food Chemistry 111 (2008) 606–615 Asp 5.21a 5. 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.72a 1.22b 5.22a 4.11b 11.11a 4.41b 37.31a 20.42a 13.72b 1.32ab 21.11a 11.82a 7.21b 34.91a 91.11b 6.41a 16.22a 5.01a 1. different numbers indicate significant differences (P < 0.32b 67.22a 15.31a 3.01b 22.93a 31.72c 3.51a Gln 8.81b 6.12b 3.32a 0.11a 51.31a 8.91a 3.61b 19.21a 40.61a 37.02a 10.01b 11.41a 20.22b 22.71a 3.01a 3.73b 3.12b 8. different letters indicate significant differences (P < 0.52a 0.52b 32.31a 17.51a 9.02b Ser 5.91a 18.02b 1.61b 3.72b 48.42a 8.71a 21.12a 2.91a 14.31a 0.01a 8.71a 3.72a 7.12b 4.52a 10.02b 17.51b 0.22b 3.71a 25.41a 4.21ab 7.02a 23.91a 24.63a 103.03a 12.81a 11.91a 11.

63b 25. Arg is responsible for unpleasant or bitter taste (Polo et al. Phe and Val.03c 1–3 : For each age and each kind of milk.01b 9..66%) and Orn acid transamination is catalyzed by aminotranferases and results and Leu to discrininant function D4 (2.01a 89. goat’s (G) or cow’s (C) milk using a thermophilic (T).82a 7.112a 46.51b 0. Amino (35%).23b 132.11b 23.71b 24. The percentage of in the formation of a-ketoacids while the a-ketoacidacceptor.92b 72.92a 26.51a 4. 60 Glu (Yvon & Rijnen. which caused the mis- other free amino acids.41ab 11. the groups presented a maximum around day 60 or 120.01b 6.91b 2.23c Asn 49.62a 9. Cys and Ala.02 23. which predominates over the proteolysis of b-casein.52b 81.62a 6.31b 14.91a 14.22a 1.62c 23. K.212a 2.02b 29.612 E.92a 60.71a 2. 4).81a 7.41a 3. .11c 14. Glu decreased with cheese ripening and classification.11c 8. different letters indicate significant differences (P < 0.42b 28. Asn of the amino acids Leu.41a 3. cheese. centroids corresponding to R.01c 3.3 mg 100 g 1 DM in 180 day old samples. a–c : For each age and each kind of culture.81a 0.41a 16.11b 1.03a 112. 1985). Four dis- criminant functions were statistically significant (P < 0.01c 112.93a 9.91b 19.12b 4.02b Glu 86.02a 14. 60 and 120 day old samples were Pro mean concentration was 13. (days) 180 Milk S G C Culture T MX M T MX M T MX M Asp 15.92b Tyr 31.08%).01b 31.33a 63.23b 5.82b 19.03a 15.41b 6.41c 92.61b 3.512b 1.42b 52.23a 7.02b 56.22ab 8.91a 3.53a 18. is transformed to the corresponding amino acid of ripening was 80. a mesophilic (M) or a mixture of thermophilic and mesophilic (MX) starter during ripening Age. 5).42a 5.C.21a 22.22a 59.71a 43.92a 6.21b 67.41b 44.51c 29. Leu and Met contributed mainly to discriminant function D2 Teleme cheese samples had high contents of Glu (Fig.01b 97.02a 34.71a 23. A DM: Dry Matter.41a 5. ripening.32a Leu 36.62a 5.51b 19.612b Gln 32.8 mg 100 g 1 DM in 1day old cheese to 11.62c Gaba 1.12a His 52.83b 30.41b 12.31b 24.92ab Ala 4. for discriminating among the stages of ripening (Fig.91a 5. Pappa.01b 34.05) between kinds of cultures.41a 36.12a 1.12c 10. often the samples that were classified into the correct group by the stage a-ketoglutarate.51a 42.32a 14.41a 18.01b 53.02a 24.02a 7.51b 30.51a 4.22a 42.21a 23.61b 1.61c 57.71c Pro 34.31a 4.31b 46.42c Cit 33.32a 27.212b AAB 34.11a 2.91c 36.01b 9. As ripening is a continuous process.41b 28. Despite the fact that the mean concentration 10 of this amino acid increased continuously throughout the ripening period (from 4.71a 3.92a 9. at 180 days of ripening.05) between kinds of milk.61b Gly 14.72a 4.62a 3.62c 6.51a 33.42a 38.912b Tau 6.01a 17. 4.52a 27.21a 21.82a 79.51a 17.11a 30.22a 5.61a 53.61a 16. Glu and Leu to discriminant function D3 (7.12a 5.23a 29.01c 105.62 34.52b 15.12b 81.82c Met 10.92b 60. 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.32a 1.312 16.02a 16.52a 34.41a 39.03a 7.02a 20.81a 0.23a 9. 2001).72b 4.52b 5.02a 108.82c 26.72a 0.01a 5.93a 63.13c Ser 16. Some overlapping between R. occurs during the first days of and Tau contributed the most to discrininant function D1(54%).51a 7.33c Val 94.81c 34.61a 5.0%.21a 5.05).53a 27.12b 1. (Fig.21a Ile 34.32b 8. 1993).61b 48.41a 50.42b Thr 31. 50 The mean concentration of Arg was 14.51b 7.41a 22.712c Phe 38.82b 37.62a 14.32b 12.83b Lys 150.63b 15.11c Cys 5.72a 3. In contrast to TFAA and most of the and 120 day old samples was observed.01b 2.82a 21.42b 30.62b 1.12b 23.71a 1.12b 25.21c 0.11b Arg 39.11b 0.01b 74.31b 5.61b 41.03a 56.2 mg 100 g 1 DM in the 1 day old cheese and reached 26.01a 70.11b 61. Stepwise discriminant analysis selected only nine amino acids Fig.53a 105.12c 7.52a 63.22a 34.91b 7.72c 9.72c 7.72b b-Ala 25. therefore it is important that this amino acid concen- 30 tration does not increase with ripening time.81a 8. 5).22a 15.61b 36.61a 20.11a 38.0 mg 100 g 1 DM in 180 day old mg/100g DM 40 cheese.43a 31.22a 5.72b 7.2 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. different numbers indicate significant differences (P < 0. 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).81b 5.21c 16.52a 62. The low concentration of Pro in Teleme cheeses could be 60 because this amino acid is normally released as a dipeptide (Prit- chard & Coolbear. %.02b 0.62c Trp 7.412a 0.01a 8.81b 7.51b 3.92a 6. 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.72c 32.42b 66.92a 5.02a 7.21a 13.6 mg 0 100 g 1 DM in 180 day old cheese). A similar decrease may not be completely separated.92c 11.31a 25.51a 6.31a 23. The distances between the group with cheese ripening was found for AAB.62a 9.52a 7.212b 45.71c Orn 14.32a 17.42a 46.42b 5. Free amino acid mean content of Teleme cheese.

(2004) found that the TFAA contents of Teleme cheeses made from ewe’s or goat’s milk did -3. Linear discriminant functions (D1 and D2) for cheese samples at 1.1 et al.001) than that of cow’s (13. in 600 Teleme cheese made with the mixed (thermophilic. 1995). on the corresponding days (Fig. 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. 7. 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.9 old cheese. 120.mesophilic) culture. -7 -4 -1 2 5 8 Orn has been found in Teleme cheeses from the first day (Table D1 (80.86%) 2). thermophilic culture.2 mg 100 g 1 DM and was significant lower (P < 0. 3. Buriana and Farag -1.05). 6. Both D1 rectly classified according to the kind of milk (100% of the total and D2 were significant (P < 0.05). Mallatou et al. The highest TFAA contents of Teleme cheeses made with the Fig. 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. regardless of the type of milk used. 7).14%) sheep’s (380 mg 100 g 1 DM) milk cheese. The Fig. In this 180 study. Tau (which origi- D2 (35.05). (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.. 5. In contrast.7 cheese (P < 0. the cheeses were cor- 180 days of ripening as well as the day cheeses entered the cold room (R). but accumulated as a metabolic product of microorganisms.001) mean concentration of Ripening day TFAA than had cheeses made with the other two types of culture. TFAA and -7 -5 -3 -1 1 3 5 Phe contributed the most to discrininant function D1 (80. Centroids as well as in ewe’s and cow’s milk cheeses (<1% of TFAA.. goat’s and cow’s milk. Low Tau contents have also been reported for Picante cheese (Freitas -1.3 (1983) found that the TFFA in ovine Teleme cheese was higher than that of bovine cheese.1 nating functions were statistically significant (P < 0. 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 < 0.44%) 2. 6 shows the evolution of TFAA for the three types of milk possesses two additional peptidases (an oligopeptidase and amino- during ripening time. AAB and GABA) did not originate from case- in. Cheeses made with the mesophilic 0 60 120 180 culture had significantly lower (P < 0.3 not differ (P > 0.01) than those of goat’s (460 mg 100 g 1 DM) or cow's D2 (19. 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. Two discrimi- -3.9 R other two types of milk. 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.001). 8).001) than that of the cheeses made from the 4. Effect of type of milk matrix and could have increased the rate of proteolysis (Chapot- Chartier.3. Cheese made with Centroids goat’s milk was also significant higher than that of sheep’s milk 0. Orn (as well as Cit.1 mg 100 g 1 DM) milk cheese. . cases).C. Both D1 and D2 were significant (P < 0. The kind of milk overall statistical analysis showed that. 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. 60.05). it is known that S. could be due to an earlier lysis of thermophilic starter bacteria that could have enhanced the early release of intracellular enzymes in cheese 3. As it can be observed in Fig. in 180 day 0. goat’s and cow’s milk.0 mg 100 g 1 DM and was signif- 1 icantly higher (P < 0.81%) while TFAA and Asn contributed mainly to discriminant function Fig. K. Stepwise discriminant analysis selected 18 amino acids for dis- criminating among the three kinds of milk (Fig. Total free amino acid mean content in cheeses made with sheep’s. 1996).14%). Tau was found in low concentrations in goat’s milk cheeses. E. 1998) and Idiazabal cheese (Barcina et al. 7. thermophilus Fig. regardless of the kind of milk and culture used).4.86%). Freitas et al.9 120 nates from Cys) is the most abundant FAA in goat’s milk. sheep's milk Generally.7 higher (P < 0. D1 (54. Orn in goat’s milk 6. 60 According to Mehaia and Al-Kanhal (1992).7 vine cheese from 297 mg 100 g 1 DM to 784 mg 100 g 1 DM.9 Ripening time cheese had a mean content of 16. Sotirakoglou / Food Chemistry 111 (2008) 606–615 613 Plot of Discriminant Functions 8.001). In ovine cheese. there was an increase in peptidase PepS) with respect to Lc. D2 (19. Moreover. Pappa.

A. T 91–109. Guinee (Eds. 17. C. 1997). M. responding to these cultures was lower than the distances corre. Kandarakis. Hannon. the mesophilic culture or the mixture of thermophilic–mesophilic culture. McSweeney. thermophilic or mesophilic adjunct culture.. G. M. Pappa. E. Lait. I. F. Proteolysis in Teleme cheese higher than that of Lc. Greek Codex Alimentarius (1998). 234–243. Prietro. cow and man. and Nunez (2001) concluded that thermophilic ad. Garde. (1984). Fresno. P.. Magboul.. goat’s or a mixture of sheep’s and goat’s milk. I. H. & Fox. Thermophilic culture 200 Moreover. Rapid analysis of amino acids Plot of Discriminant Functions using precolumn derivatisation. M.. 93–104. Izco. E. 1990. cheeses made with cow’s milk had higher TFAA contents Thermophilic-Mesophilic culture than had cheeses made with ewe’s milk. Influence of criminating functions were statistically significant (P < 0. Cheeses made with the mixed ther- mophilic–mesophilic culture ranged over intermediate levels. 73. P. Free amino acids by high performance were successfully clustered by the type of starter. 86. International Dairy Journal. A. composition and sensory characteristics of Teleme cheese during ripening.. Cogan. Protection. M. Influence of starters on chemical.). 599–610. (1979). 3479–3484.C. & Nunez. International Dairy Journal. F. M.1 Picante cheese.. P.. FEMS Microbiology Reviews. 273–290. Fox. J. J. H. Pappa... F. Ramos.. L. 9). 88. . (2001). N.. T.05). P. levels (Khalid & Marth. Cohen.05). J. Production of Cheddar cheese 3. Fig. & T. Tzanetakis. S. J. 31. B. & Steele. 27–42).. A. Buriana. Le observed (Fig.. & Coolbear. L. (1990). & Torre. 336. Oumer. A. Cheese: chemistry. TFAA and Leu contributed the most to discrininant function D1 570–581. N.. L. Les autolysins des bactéries lactiques. Les acides aminés du fromage Télémé. Athens. Hayaloglou. In P. (2004). S. A. Upadhyay. while TFAA and Asn contributed mainly to discriminant Pappa.46%).. (2006a). types of milk and culture on the manufacturing practices. & Farag. E. Two dis.. Both D1 and D2 were significant (P < 0.. Anifantakis. physics and microbiology (3rd ed. C. (2005). J. 732–739. R. D2 (5. USA: Waters Association. & Sotirakoglou. Influence of starter cultures on the proteolysis of Teleme cheese made from different types of milk. K. Gaya. J. & McSweeney. The variation of amino acids in Telemea cheese 5.. L. (FIL- thermophilic (T). Franco. Xavier Malcata.54%). M. (94. Cheese and processed cheese products. Food Control. Fig. 12. A.. The percentage of the samples classified into (2006b). 47. Journal of Dairy Science.. Tomillo. 179–206.. P. 14. the Khalid. 4). Pritchard. International peptidases and because their peptidases have higher expression Dairy Journal.. 977–987. E. Garde. although some liquid chromatography and peptides by gel electrophoresis in Mahon cheese overlapping between mixed and mesophilic cultures was during ripening. The total free amino acid (TFAA) content increased 600 with ripening time. Barcina. H. G. M. Journal of Dairy Science. 351–353. 14. K. 9. (1981). (1998). 50. L.. (2000). culture. goat. M. when inoculated into milk. Proteolysis in Hispanico cheese manufactured using a mesophilic starter. M. C. A. & Vlachos. (1992). 81–86. B. Kandarakis. J. 161–164. the kind of culture added and the ripening time. K. (1985). Greek cheeses – A tradition of centuries. 9). 88140. showed considerably pp. junct culture. The TFAA contents were highest in cheeses made with the thermophilic culture and lowest in cheeses made 400 with the mesophilic culture. P. M. Characterization of volatile flavour compounds in Roncal cheese extracted by the purge and trap method and analyzed by GC–MS. Garde.. (1993). criminating among the three types of culture (Fig.. M. E.9 adjunct culture. E. Determination of dry matter. ture. culture Chapot-Chartier.. J. M.. The physiology and biochemistry of the sponding to thermophilic culture from mixed or mesophilic proteolytic system in lactic acid bacteria. 76. Also. A. A.. & Carballo. Teleme cheese from deep-frozen curd.. Medina. G. 5. & Vafopoulou. M. GR: National Printing Office. Operator’s Manual No. (1995). The distance between the group centroids cor- Lait. and Nunez (2002) found simi. Official journal of the hellenic republic Vol. Committee of Greece (pp.. L. (1995). lactis INIA 415 -0. T. Journal of Dairy Science. M. & Sanchez. Y. The effects of cultivating lactic acid cultures with bacteriocin-producing lactic acid bacteria. a mesophilic (M) or a mixture of thermophilic–mesophilic (MX) IDF standard no. Sasaki. & Tan. (1983).. P. K. F. a thermophilic starter and bacteriocin-producing Lactococcus lactis subsp. lar results when they manufactured Hispanico cheese with MA. Bosman. Medina. L. S.. cremoris SK11 derivative with enhanced M aminopeptidase activity. 53–59. I.. Fox. & Al-Kanhal. E. Anifantakis. 391–433). Garde. F. Tomillo.. A. Proteolysis in cheese during ripening. A. Linear discriminant functions (D1 and D2) for cheese samples made using a IDF (1986). In National Dairy culture. 85–96. & Zerfiridis. I. Food Chemistry.. A. Gaya. I.. Oumer. PepN and PepC (Rul & Monnet. Fox. H. H. Food Control. V. A. (2004).1 during pickling. 70. activities of PepX. Johnson. (1996). overall proteolytic activity of lactobacilli has been found to be Mallatou. & Tarvin.9 Proteolysis in Turkish white-brined cheese made with defined strains of -6 -4 -2 0 2 4 6 Lactococcus. G. & Ordonez.. Food Control. 11. Lait.. Medina. Food Chemistry. Polychroniadou. (1991). (2004).. A. Sotirakoglou / Food Chemistry 111 (2008) 606–615 1000 4. & Boumba. Midford. McSweeney. P. 8.46%) Hayaloglou. Egyptian Journal of Dairy Science. Journal of Agricultural and Food Chemistry. Conclusions Total amino acids mg/100g DM 800 The free amino acid content of Teleme cheese was considerably affected by the kind of milk used. biochemical and sensory changes in Turkish white-brined cheese during ripening. A. (1984). 64. 2669–2684. P. Cheeses Polo. Ibanez. 16. V.. & Marth. the correct group by the type of culture was 88. (2002). F. 6. Guven. A.1 using a Lactococcus lactis ssp. E. Taurine and other free amino acids in milk higher aminopeptidase activity than did mesophilic adjunct cul- of camel. because lactobacilli possess additional made from sheep’s. 3460–3474.. Milchwissenschaft. A. Bidlingmeyer.. Journal of Chromatography. Influence of milk source and ripening time on free amino acid profile of 1. culture. 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