Composition containing amino acids and process for producing same

Abstract

The invention relates to a process for the preparation of a composition including free amino acids, oligopeptides and polypeptides, characterized in that the process comprises at least the following step: adding at least two different peptidases, which are present in purified form, to a composition comprising at least one protein.

Claims

1. A process for producing a composition comprising free amino acids, oligopeptides, and polypeptides, comprising: adding at least one purified microbial exopeptidase and at least one purified microbial endopeptidase to a composition comprising at least one protein; and incubating the at least one exopeptidase, the at least one endopeptidase and the composition at a temperature of 40 C.-60 C.; wherein: prior to adding the at least one purified microbial exopeptidase and at least one purified microbial endopeptidase, the concentration of the at least one protein in the composition is 5-30 wt % based on the total weight of the composition; prior to adding the at least one purified microbial exopeptidase and at least one purified microbial endopeptidase, the pH of the composition is 4-6; and the amount of the at least one purified microbial exopeptidase and at least one purified microbial endopeptidase added to the composition is 2-10 grams of each peptidase per 100 grams of the at least one protein.

2. The process according to claim 1, wherein the microbial origin of the at least one exopeptidase and the at least one endopeptidase is bacterial or fungal.

3. The process according to claim 1, wherein prior to adding at least one exopeptidase and the at least one endopeptidase, the concentration of the at least one protein in the composition is 10-20 wt % based on the total weight of the composition.

4. The process according to claim 1, wherein the incubating occurs 1-24 hours.

5. The process according to claim 1, wherein the incubating temperature is 45 C.-60 C.

6. The process according to claim 1, wherein, prior to adding the at least one exopeptidase and the at least one endopeptidase, the pH of the composition is 4.5-6.

7. The process according to claim 1, further comprising adding calcium carbonate and/or magnesium carbonate.

8. The process according to claim 1, wherein the at least one protein in the composition is selected from the group consisting of whey, a whey protein concentrate, a milk protein concentrate, a slurry from food production, a distillery slurry, a brewery slurry, pea, lentil, bean, soybean, and combinations of two or more thereof.

9. The process according to claim 1, wherein the amount of each of the at least one exopeptidase and the at least one endopeptidase added is 500-20,000 U per 100 grams of the at least one protein in the composition.

10. The process according to claim 1, wherein at least one of the at least one purified microbial exopeptidase and the at least one purified microbial endopeptidase is added repeatedly to the composition.

11. The process according to claim 10, wherein the at least one of the at least one exopeptidase and the at least one endopeptidase is repeatedly added in equal amounts and at equal time intervals.

12. The process according to claim 1, wherein the at least one endopeptidase is added prior to adding the at least one exopeptidase.

13. The process according to claim 1, wherein the composition is first incubated with the at least one endopeptidase for 1-10 hours, and then incubated with the at least one exopeptidase for 3-40.

14. The process according to claim 13, wherein the at least one endopeptidase and/or the at least one exopeptidase is/are added in equal amounts at equal time intervals.

15. A composition produced by the process according to claim 1, comprising free amino acids, oligopeptides, and polypeptides, wherein: 0.5-25 wt % of the composition is free amino acids, based on a total weight of free amino acids, oligopeptides, polypeptides, and proteins without the added peptidases; and 25-65 wt % of the composition is oligopeptides, excluding the wt % of the at least one exopeptidase and the at least one endopeptidase.

16. The composition according to claim 15, wherein the composition comprises from 0.3-13 wt % of the free amino acids and the free amino acids are selected from the group consisting of isoleucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, and combinations thereof.

17. The composition according to claim 15, wherein 0-40 wt % of the at least one protein is present as uncleaved protein.

18. The composition according to claim 15, wherein the composition has a pH of 3-7.

19. The composition according to claim 15, further comprising magnesium and calcium in a weight ratio of 1.25:1-3:1 magnesium:calcium.

20. A food product comprising the composition according to claim 15, wherein the food product is selected from the group consisting of a fruit juice beverage, a milk beverage, a whey beverage, a yogurt, an ayran, and a kefir.

21. A personal care product comprising the composition according to claim 15, wherein the care product is a soap, a shampoo, a hair treatment, a hair mask, a hair pack, a hair tonic, a conditioner, a bath essence, a body lotion, a body gel, a lotion for treating rosacea, a cream for treating acne, a facial mask or a facial cream.

22. The food product according to claim 20, wherein a total content of the free amino acids, the oligopeptides, and the polypeptides is from 2 wt % to 25 wt % of the food product.

23. The food product according to claim 20, wherein a total content of the free amino acids, the oligopeptides, and the polypeptides in the food product is from 5 to 50 grams.

Description

EXAMPLES

Example 1

(1) Whey protein concentrate WPC 80 with a dry matter content of 25% and a protein content of 80% in the dry matter was diluted 1:2 with water. The protein content of the solution thus produced was 10 wt %, based on the total weight of the solution produced.

(2) The pH of this solution was adjusted to 5.5 with citric acid.

(3) Two test preparations of 1 liter (100 g protein) each were produced. 2 g of an endopeptidase with an activity of about 1000 SAPU/g were added to each of the two preparations at the beginning and five more times in one-hour intervals. The total quantity of added endopeptidase was 12 g/100 g source protein in each case.

(4) The preparations were incubated at 50 C.

(5) After five hours, a 50 ml sample was taken from each preparation (samples I5 and II5comparison example). The preparations were further incubated at 50 C. overnight.

(6) The next morning, 2 g of an exopeptidase with about 900 CPGU were added to each of the two preparations five times in two-hour intervals. The total quantity of added exopeptidase was 10 g/100 g source protein in each case. The preparations were further incubated at 50 C.

(7) Another 50 ml sample was then taken from each preparation (samples I9 and II9), and all four samples were analyzed.

(8) To this end, the samples were homogenized. 0.5 g-2.5 g of the homogenized samples were weighed to the nearest 0.1 mg into a Falcon tube. 1 ml norleucine solution (0.1312 g/200 ml) were added as an internal standard (IS) and topped up to 25 ml with 3% 5-sulfosalicylic acid. The tubes were placed in a shaker and shaken vigorously for 20 minutes. They were then centrifuged at 5000 rpm for 5 minutes. The supernatant was filtered through a syringe filter (0.45 m). 0.5 ml of the filtrate and 0.5 ml sample buffer for chromatography were pipetted into a reaction vessel, which was then sealed. The solution thus produced was used for chromatographic analysis on the amino acid analyzer.

(9) In parallel, the amino acid composition of the fraction containing free amino acids and oligopeptides was determined. To this end, 0.5 g-2.5 g of the homogenized sample were again weighed to the nearest 0.1 mg into a Falcon tube. 2.5 ml norleucine solution (0.1312 g/200 ml) were pipetted in as an internal standard (IS) and topped up to 25 ml with 3% 5-sulfosalicylic acid. The tubes were shaken and centrifuged in the manner described above; the water-insoluble polypeptides and proteins contained in the precipitate were discarded. 1 ml of the supernatant was pipetted into a 50 ml glass digestion vessel, mixed with 9 ml 6N hydrochloric acid, and the vessels were tightly sealed with a screw cap. The vessels were placed in a heating block and the sample solution was digested for 8 h at 105 C. 2 ml of the digested solution were evaporated to dryness. The residue was taken up in 1 ml sample buffer for chromatography and transferred to a reaction vessel, which was then sealed. The solution thus produced was used for chromatographic analysis on the amino acid analyzer. In addition, the content of oligopeptides in the composition can be calculated by subtracting the content of free amino acids determined in step 1.

(10) Table 1 shows the composition of the four samples that were analyzed.

(11) TABLE-US-00001 TABLE 1 Sample composition Results free amino acids: Sample I 5 Sample II 5 Sample I 9 Sample II 9 mg/kg test preparation essential amino acids (proteinogenic amino acids): Threonine (Thr) 57 56 709 738 Valine (Val) 112 111 977 949 Methionine (Met) 75 85 135 142 Isoleucine (Iso) 49 44 344 361 Leucine (Leu) 947 853 >2800 >2800 Phenylalanine (Phe) 175 143 843 815 Lysine (Lys) 1279 1209 >2800 >2800 Tryptophan (Trp) not determined semi-essential amino acids (proteinogenic amino acids): Histidine (His) 74 68 384 384 Arginine (Arg) 264 247 795 791 non-essential amino acids (proteinogenic amino acids): Aspartic acid (Asp) 25 23 390 378 Serine (Ser) 60 57 395 404 Asparagine (Asn) 49 30 343 292 Glutamic acid (Glu) 308 248 1591 1588 Glycine (Gly) <10 <10 116 124 Alanine (Ala) 105 93 634 665 Cysteine (Cys) not determined Tyrosine (Tyr) 148 142 814 818 Proline (Pro) 29 40 104 125 Glutamine (Gln) not determined other determined substances Taurine (Tau) not not not not analyzable analyzable analyzable analyzable Citrulline (Cit) <10 <10 34 41 Cystine (Cys2) 186 189 341 348 Cystathionine 191 173 not not (Cystha) analyzable analyzable 3-methyl-histidine <10 <10 <10 <10 (3Mehis) 1-methyl-histidine <10 <10 <10 <10 (1Mehis) Hydroxylysine <10 <10 <10 <10 (Hylys) Ornithine (Orn) <10 <10 <10 <10 Hydroxyproline <10 <10 <10 <10 (Hypro) Results NPN extract (oligopeptides and free amino acids: Sample I 5 Sample II 5 Sample I 9 Sample II 9 mg/kg test preparation essential amino acids (proteinogenic amino acids): Threonine (Thr) 4813 4641 5223 5266 Valine (Val) 2583 2672 3069 3287 Methionine (Met) 791 154 533 223 Isoleucine (Iso) 2968 2840 3410 3701 Leucine (Leu) 5909 5696 6864 6944 Phenylalanine (Phe) 1590 1564 1890 1846 Lysine (Lys) 5788 5490 6544 6530 Tryptophan (Trp) not determined semi-essential amino acids (proteinogenic amino acids): Histidine (His) 1081 1004 1239 1190 Arginine (Arg) 1300 1185 1360 1285 non-essential amino acids (proteinogenic amino acids): Aspartic acid (Asp) 5459 5145 not 2004 analyzable Serine (Ser) 3344 2694 3394 3249 Asparagine (Asn) nn nn nn nn Glutamic acid (Glu) 11235 9516 9333 8550 Glycine (Gly) 941 873 1038 975 Alanine (Ala) 3279 3123 3225 3176 Cysteine (Cys) not determined Tyrosine (Tyr) 1122 860 1563 1420 Proline (Pro) 4722 4722 4644 4522 Glutamine (Gln) not determined other determined substances Taurine (Tau) not not not not analyzable analyzable analyzable analyzable Citrulline (Cit) nn nn nn nn Cystine (Cys2) 747 488 1374 1160 Cystathionine nn nn nn nn (Cystha) 3-methyl-histidine nn nn nn nn (3Mehis) 1-methyl-histidine nn nn nn nn (1Mehis) Hydroxylysine nn nn nn nn (Hylys) Ornithine (Orn) nn nn nn nn Hydroxyproline nn nn nn nn (Hypro)

(12) After hydrolysis was complete, about 50 to 60% oligopeptides and about 15% free amino acids, divided into about 10% essential amino acids and 5% non-essential amino acids, in each case based on the total weight of the source protein, in this case 100 g protein, were present in the preparations that were incubated with the endo- and exopeptidases.

(13) The pH, the lactose content, and taste of the four samples were also assayed. The results are summarized in Table 2.

(14) TABLE-US-00002 TABLE 2 Taste analysis of the generated hydrolysates: Sample Sample Sample Sample WPC I 5 II 5 I 9 II 9 80 1:2 pH 4.97 5.03 5.04 5.10 Taste not bitter not bitter not bitter not bitter Lactose 0.9 0.8 [g/100 g]

Example 2

(15) Whey protein concentrate WPC 80 with a dry matter content of 25% and a protein content of 80% in the dry matter was diluted 1:2 with water. The protein content of the solution thus produced was 9.98 wt %, based on the total weight of the solution produced.

(16) The pH of this solution was originally 6.32 and was then adjusted to 5.45 with citric acid.

(17) The preparation volume was 2 liters in each case. An endopeptidase with an activity of about 1000 SAPU/g and an exopeptidase with about 900 CPGU were added. Enzymatic hydrolysis took place at 50 C.

(18) Table 3 shows the hydrolysis scheme:

(19) TABLE-US-00003 TABLE 3 Hydrolysis scheme Day Time Preparation EZ 40 Preparation EZ 41 Day 1 7:05 am 2 g endopeptidase 2 g endopeptidase 9:05 am 2 g endopeptidase 2 g endopeptidase 2 g exopeptidase 2 g exopeptidase 11:10 am 2 g endopeptidase 2 g endopeptidase 2 g exopeptidase 2 g exopeptidase 2:10 pm End 4 g endopeptidase 4 g exopeptidase Day 2 9:00 am End Total quantity of 3 g 5 g endopeptidase [g/100 g source protein] Total quantity of 2 g 4 g exopeptidase [g/100 g source protein]

(20) Samples were then taken and analyzed in the manner described in Example 1.

(21) Table 4 shows the composition of the analyzed samples.

(22) TABLE-US-00004 TABLE 4 Sample composition Results NPN extract Results (oligopeptides free amino acids: and free amino acids: EZ40 EZ41 EZ40 EZ41 mg/kg test preparation essential amino acids (proteinogenic amino acids) Threonine (Thr) 143 434 3782 4482 Valine (Val) 166 513 2222 2694 Methionine (Met) 114 257 871 978 Isoleucine (Iso) 101 283 2458 2956 Leucine (Leu) 808 <2000* 4643 >5500* Phenylalanine (Phe) 143 354 1217 1486 Lysine (Lys) 1147 <2000* <4000* >4000* Tryptophan (Trp) not determined semi-essential amino acids (proteinogenic amino acids) Histidine (His) 124 253 727 949 Arginine (Arg) 256 536 956 1214 non-essential amino acids (proteinogenic amino acids) Aspartic acid (Asp) 46 163 4136 5076 Serine (Ser) 125 289 2557 2957 Asparagine (Asn) 61 138 <50 <50 Glutamic acid (Glu) 261 666 >8000* >8000* Glycine (Gly) 19 48 685 878 Alanine (Ala) 124 381 2637 >3000* Cysteine (Cys) not determined Tyrosine (Tyr) 193 515 1064 1470 Proline (Pro) 21 79 >4000* >4000* Glutamine (Gln) not determined other determined substances Taurine (Tau) 68 83 87 90 Citrulline (Cit) <10 <10 <50 <50 Cystine (Cys2) 103 261 399 516 Cystathionine <10 <10 <50 <50 (Cystha) 3-methyl-histidine <10 <10 <50 <50 (3Mehis) 1-methyl-histidine <10 <10 <50 <50 (1Mehis) Hydroxylysine <10 <10 <50 <50 (Hylys) Ornithine (Orn) <10 <10 <50 <50 Hydroxyproline <10 <10 <50 <50 (Hypro)

(23) The samples were also analyzed in terms of phase separation, pH, and taste. The results are summarized in Table 5.

(24) TABLE-US-00005 TABLE 5 Taste analysis of the generated hydrolysates: Sample EZ 40 Sample EZ 41 Phase separation no yes pH 4.50 4.47 Taste bitter bitter

Example 3

(25) Whey protein concentrate WPC 80 with a dry matter content of 25% and a protein content of 80% in the dry matter was used diluted with water or undiluted.

(26) Table 6 shows the composition of the test preparations with their pH values. A pH adjustment was not necessary.

(27) TABLE-US-00006 TABLE 6 Composition of the test preparations EZ 42 EZ 43 EZ 44 EZ 45 Protein content [%] 11.27 20.43 11.27 10.00 pH 5.33 5.34 5.33 5.60 Preparation volume [l] 2 2 2 1

(28) An endopeptidase with an activity of about 1000 SAPU/g and an exopeptidase with about 900 CPGU were added. Enzymatic hydrolysis took place at 50 C.

(29) Table 7 shows the hydrolysis scheme.

(30) TABLE-US-00007 TABLE 7 Hydrolysis scheme: Day Time EZ 42 EZ 43 EZ 44 1 10:00 am 2 g endop. 2 g endop. 10 g endop. 11:00 am 2 g endop. 2 g endop. 12:00 noon 2 g endop. 2 g endop. 1:00 pm 2 g endop. 2 g endop. 2:00 pm 2 g endop. 2 g endop. 4:00 pm End of approach no. 1 2 7:00 am End of approach no. 2 7:15 am 2 g exop. 2 g exop. 8 g exop. 8:15 am 2 g exop 2 g exop 9:15 am 2 g exop 2 g exop 10:15 am 2 g exop. 2 g exop 12:15 pm End of approach no. 3 Day Time EZ 45 1 7:15 am 2.5 g endop. 1:15 pm End of approach no. 1 2 8:45 am End of approach no. 2 7:30 am 2 g exop. 12:30 pm End of approach no. 3

(31) Samples were then taken and analyzed in the manner described in example 1.

(32) Table 8 shows the composition of the analyzed samples.

(33) TABLE-US-00008 TABLE 8 Sample composition Results free amino acids: EZ 42 EZ 43 6 h 21 h 26 h 6 h 21 h 26 h mg/kg test preparation essential amino acids (proteinogenic amino acids) Threonine (Thr) 85 267 671 71 263 992 Valine (Val) 60 205 631 96 229 818 Methionine (Met) 150 341 453 203 415 632 Isoleucine (Iso) 39 156 398 70 186 465 Leucine (Leu) 522 1351 2536 582 1407 3049 Phenylalanine (Phe) 126 335 709 165 354 886 Lysine (Lys) 1102 1731 2898 1598 2126 3723 Tryptophan (Trp) not determined not determined semi-essential amino acids (proteinogenic amino acids) Histidine (His) 68 164 303 74 150 400 Arginine (Arg) 230 408 421 220 448 359 non-essential amino acids (proteinogenic amino acids) Aspartic acid (Asp) 33 134 464 40 121 574 Serine (Ser) 69 169 368 70 170 613 Asparagine (Asn) nn 164 386 39 99 609 Glutamic acid (Glu) 141 614 1909 194 553 2305 Glycine (Gly) 26 63 133 30 67 195 Alanine (Ala) 85 239 554 112 248 751 Cysteine (Cys) not determined not determined Tyrosine (Tyr) 64 260 626 58 251 681 Proline (Pro) nn 76 150 nn 72 236 Glutamine (Glu) not determined not determined other determined substances Taurine (Tau) not not not not not not analyzable analyzable analyzable analyzable analyzable analyzable Citrulline (Cit) nn nn 156 nn nn 431 Cystine (Cys2) nn 62 156 nn nn 431 Cystathionine 89 151 68 115 145 63 (Cystha) 3-methyl-histidine nn nn nn nn nn nn (3Mehis) 1-methyl-histidine nn nn nn nn nn nn (1Mehis) Hydroxylysine nn nn nn nn nn nn (Hylys) Ornithine (Orn) nn 19 55 nn nn 96 Hydroxyproline nn nn nn nn nn nn (Hypro) Total 2889 6909 14039 3755 7353 17935 Results NPN-extract (oligopeptides and free amino acids): EZ 42 EZ 43 6 h 21 h 26 h 6 h 21 h 26 h mg/kg test preparation essential amino acids (proteinogenic amino acids) Threonine (Thr) 4151 5074 4963 5574 5355 1927 Valine (Val) 4016 3643 3687 5923 5821 5951 Methionine (Met) 423 768 755 1098 1073 307 Isoleucine (Iso) 3953 3864 4059 6293 6110 6356 Leucine (Leu) 8149 7809 7971 12565 12778 13637 Phenylalanine (Phe) 2316 2175 2261 3186 3285 3533 Lysine (Lys) 7261 6975 7228 10794 10943 12236 Tryptophan (Trp) not determined not determined semi-essential amino acids (proteinogenic amino acids) Histidine (His) 1536 1327 1544 2057 2103 2547 Arginine (Arg) 2308 1487 1321 2253 2599 1666 non-essential amino acids (proteinogenic amino acids) Aspartic acid (Asp) 7188 6941 7287 10038 10948 10159 Serine (Ser) 2344 3104 3039 3161 2787 798 Asparagine (Asn) nn nn nn nn nn nn Glutamic acid (Glu) 12268 12614 12217 15289 16814 7777 Glycine (Gly) 1763 1274 1386 1800 2119 2186 Alanine (Ala) 4958 4383 4159 7320 7204 7185 Cysteine (Cys) not determined not determined Tyrosine (Tyr) 938 1316 1611 1522 1463 269 Proline (Pro) 5506 5271 4993 8859 8653 9694 Glutamine (Glu) not determined not determined other determined substances Taurine (Tau) not not not not not not analyzable analyzable analyzable analyzable analyzable analyzable Citrulline (Cit) nn nn 130 nn nn nn Cystine (Cys2) 1898 1175 1617 2515 3922 3235 Cystathionine nn nn nn nn nn nn (Cystha) 3-methyl-histidine nn nn nn nn nn nn (3Mehis) 1-methyl-histidine nn nn nn nn nn nn (1Mehis) Hydroxylysine nn nn nn nn nn nn (Hylys) Ornithine (Orn) nn nn 59 nn nn 260 Hydroxyproline nn nn nn nn nn nn (Hypro) Total 70976 69191 70287 100247 103977 89723 Results free amino acids: EZ 44 EZ 45 6 h 21 h 26 h 6 h 25.5 h 30.5 h mg/kg test preparation essential amino acids (proteinogenic amino acids) Threonine (Thr) 76 463 695 35 385 517 Valine (Val) 49 417 612 11 364 471 Methionine (Met) 170 398 532 100 286 317 Isoleucine (Iso) 33 246 481 34 220 301 Leucine (Leu) 576 1798 2731 237 1142 1592 Phenylalanine (Phe) 146 498 757 93 347 479 Lysine (Lys) 1141 2192 2961 691 1350 1831 Tryptophan (Trp) not determined not determined semi-essential amino acids (proteinogenic amino acids) Histidine (His) 76 208 330 36 128 209 Arginine (Arg) 219 462 406 126 340 394 non-essential amino acids (proteinogenic amino acids) Aspartic acid (Asp) 48 251 232 28 93 156 Serine (Ser) 61 257 384 39 219 278 Asparagine (Asn) nn 220 356 nn 146 154 Glutamic acid (Glu) 156 1070 2099 118 924 1382 Glycine (Gly) 10 94 152 nn 63 101 Alanine (Ala) 93 369 606 66 291 431 Cysteine (Cys) not determined not determined Tyrosine (Tyr) 80 382 684 27 250 412 Proline (Pro) nn 94 173 nn nn 194 Glutamine (Glu) not determined not determined other determined substances Taurine (Tau) not not not not not not analyzable analyzable analyzable analyzable analyzable analyzable Citrulline (Cit) nn 58 105 nn nn 58 Cystine (Cys2) 27 59 228 nn 23 55 Cystathionine 48 89 136 31 48 nn (Cystha) 3-methyl-histidine nn nn nn nn nn nn (3Mehis) 1-methyl-histidine nn nn nn nn nn nn (1Mehis) Hydroxylysine nn nn nn nn nn nn (Hylys) Ornithine (Orn) nn 13 146 nn 29 54 Hydroxyproline nn nn nn nn nn 51 (Hypro) Total 3009 9638 14806 1672 6648 9437 Results NPN-extract (oligopeptides and free amino acids): EZ 44 EZ 45 6 h 21 h 26 h 6 h 25.5 h 30.5 h mg/kg test preparation essential amino acids (proteinogenic amino acids) Threonine (Thr) 5627 2394 6272 2782 4669 3677 Valine (Val) 3457 3624 3881 3236 3310 3361 Methionine (Met) 950 455 1300 444 714 510 Isoleucine (Iso) 3503 3849 4129 3443 3566 3598 Leucine (Leu) 7474 7984 8521 6684 6916 7196 Phenylalanine (Phe) 2091 1891 2503 1721 1955 1994 Lysine (Lys) 6759 7179 7806 5881 5975 6121 Tryptophan (Trp) not determined not determined semi-essential amino acids (proteinogenic amino acids) Histidine (His) 1247 1419 1695 1115 1229 1327 Arginine (Arg) 1567 1349 1336 1264 1444 1332 non-essential amino acids (proteinogenic amino acids) Aspartic acid (Asp) 6711 6333 8226 5793 6098 6405 Serine (Ser) 3763 1070 4374 1523 2960 2174 Asparagine (Asn) nn nn nn nn nn nn Glutamic acid (Glu) 13549 10107 14520 10546 11651 10480 Glycine (Gly) 1163 1324 1571 1010 1114 1208 Alanine (Ala) 4496 4273 4609 4061 3880 3918 Cysteine (Cys) not determined not determined Tyrosine (Tyr) 1379 545 1995 546 1322 1086 Proline (Pro) 5490 5579 5819 5146 5035 4910 Glutamine (Glu) not determined not determined other determined substances Taurine (Tau) not not not not not not analyzable analyzable analyzable analyzable analyzable analyzable Citrulline (Cit) nn nn nn nn nn nn Cystine (Cys2) 742 1765 1276 1207 1059 1583 Cystathionine 48 89 136 31 48 nn (Cystha) 3-methyl-histidine nn nn nn nn nn nn (3Mehis) 1-methyl-histidine nn nn nn nn nn nn (1Mehis) Hydroxylysine nn nn nn nn nn nn (Hylys) Ornithine (Orn) nn nn 139 nn nn 64 Hydroxyproline nn nn nn nn nn nn (Hypro) Total 69968 61140 79972 56402 62897 60944

(34) The samples were analyzed specifically in terms of their taste. The samples were not bitter.