(Development of natural seasoning kokumi using domestic agricultural products)

(Development of natural seasoning kokumi using domestic agricultural products)

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Natural or artificial flavor Reaction flavor 단백가수분해물, 효모추출물등 ( 펩타이드, 아미노산류맛 ) kokumi 기본맛 ( 단맛, 신맛, 짠맛, 쓴맛, 감칠맛 ) - 28 -

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종류 생산 ( 톤 / 년 ) 종류 생산 ( 톤 / 년 ) 양파엑기스 1,500 양배추엑기스 700 당근엑기스 900 마늘엑기스 500-30 -

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우리밀 제형 수세 Amylase Protease 추출 분말 - 34 -

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β - 37 -

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α - 39 -

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No Name Protease activity(u/ml) 1 SEM-11 64 2 SEM-27 68 3 SEM-54 65 4 SEM-61 69 5 SEM-87 59 6 SEM-93 77 7 SEM-130 73 8 SEM-133 89 9 SEM-139(Y균 ) 70 10 SEM-140 41 11 SEM-156 41 12 SEM-158 43 13 SEM-162 57 14 SEM-166 50 15 SEM-176 43 16 SEM-177 46 Fungi Bacteria - 43 -

Enzyme activity Selected strain Aspergillus sp. SP1 Bacillus sp. SP1 Protease (U/ml) 154(U/ml) 118(U/ml) - 44 -

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A B - 50 -

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우리밀 분쇄 수세 Amylase Protease 추출 분말 - 53 -

M wheat 10 30 60 120 180 M min 107,000 81,000 203,646 116,134 92,266 48,700 50,400 33,800 27,000 37,043 28,920 20,700 20,081 M wheat 5h 10h 15h 20h M 107,000 81,000 48,700 33,800 27,000 203,646 116,134 92,266 50,400 37,043 28,920 20,700 20,081 6,936-54 -

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마늘, 양파 탈피 분쇄 열추출 여과 농축 분말 - 56 -

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100 30 80 60 40 25 20 15 10 Protoplast format 20 0 0 0 20 40 60 80 100 120 140 Westase treatment time(min) 5 Protoplast regera - 58 -

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Xn Independent Level variables -α -1 0 +1 +α X1 glucose 33.18 40.00 50.00 60.00 66.82 X2 yeast 29.09 32.50 37.50 42.50 45.91 extract X3 ammonium phosphate 4.95 7.00 10.00 13.00 15.05 α - 60 -

α α Xn Independent Level variables -α -1 0 +1 +α X1 glucose 53.18 60.00 70.00 80.00 86.82 X2 yeast 15.68 22.50 32.50 42.50 49.32 extract X3 ammonium phosphate 1.95 4.00 7.00 10.00 12.05 α - 61 -

α α - 62 -

Factor Name Level Low level High Level A glucose 80 60 80 B yeast extract 37.5 22.5 42.5 C Ammonium phosphate 4 4 10 Response Prediction SE Mean 95% CI low 95% CI high RNA content 4.152 0.073 3.979 4.325 Culture time(h) 0 4 10 16 24 Glutathione (ug/ml) 8 12 36 68 52-63 -

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1.8 6 1.6 1.4 5 1.2 4 OD600 1.0 0.8 0.6 0.4 0.2 0.0 0 5 10 15 20 3 2 1 0 RNA(mg/ml), ph Culture time(h) - 67 -

1 2 3 Inlet temp(. outlet temp( rpm 170 70 18,000 170 80 18,000 170 90 18,000 175 70 18,000 175 80 18,000 175 90 18,000 180 70 18,000 180 80 18,000 180 90 18,000-68 -

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μ μ μ - 72 -

μ μ - 73 -

μ μ μ - 74 -

μ μ - 75 -

μ - 76 -

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μ μ μ - 78 -

μ μ - 79 -

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μ μ - 81 -

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μ μ μ - 83 -

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RI A) Compounds P.A (%) ID B) Aldehydes and Ketones 829 Furfural 0.66 MS/RI 953 Benzaldehyde 3.52 MS/RI 1035 Benzeneacetaldehyde 23.02 MS/RI 1462 2,6-Di(t-butyl)-4-hydroxy-4-methyl-2,5-cyclohexadien-1-one 1.68 MS 1481 5-Methyl-2-phenyl-2-hexenal 17.60 MS/RI 1499 2,3,6,7-Tetrahydrocyclopent[b]azepine-8[1H]-one 1.18 MS Cyclic compounds 1625 [1-Butylheptyl]-benzene 0.36 MS 1868 Cyclohexadecane 2.44 MS Furans 855 2-Furanmethanol 3.76 MS/RI S-Containing compounds 903 Methional 0.67 MS/RI 963 Dimethyl trisulfide 1.05 MS/RI 1195 1,4-Dimethyl tetrasulfide 0.60 MS/RI 1401 3-Phenylthiophene 1.08 MS N-Containing compounds 990 2-Ethyl-6-methylpyrazine 0.82 MS/RI 1235 2-Isoamyl-6-methylpyrazine 0.59 MS 1282 Indole 6.88 MS/RI 1449 3-Phenylpyridine 1.25 MS 1902 Hexahydro-3-[2-methylpropyl]-pyrrolo[1,2-a]pyrazine-1,4-dione 14.30 MS Aliphatic and Terpene Hydrocarbons - 85 -

958 2,6,6-Trimethyl-bicyclo[3,1,1]hept-2-ene 1.52 MS/RI 986 β-myrcene 2.30 MS/RI 999 l-limonene 7.78 MS/RI 1289 Tridecane 0.61 MS/RI 1490 Pentadecane 0.47 MS/RI 1589 Hexadecane 1.32 MS/RI 1689 Heptadecane 0.73 MS/RI 1782 1-Octadecene 1.23 MS/RI 1987 Eicosane 0.89 MS/RI 1889 Nonadecane 0.70 MS/RI Miscellaneous compounds 1302 2-Methoxy-4-vinylphenol 0.46 MS/RI 1510 Tributyl phosphate 0.51 MS - 86 -

RI A) Compounds P.A (%) ID B) Aldehydes and Ketones 954 Benzaldehyde 5.97 MS/RI 1036 Benzeneacetaldehyde 19.84 MS/RI 1146 3-Phenylpropenal 1.70 MS/RI 1464 2,6-Di-[t-butyl]-4-hydroxy-4-methyl-2,5-cyclohexadien-1-one 1.36 MS Cyclic compounds 865 1,3-Dimethylbenzene 4.54 MS/RI 926 2-Cyclohexen-1-one 1.32 MS/RI 973 1-Ethyl-2-methylbenzene 0.21 MS/RI 1057 Acetophenone 2.33 MS/RI 1723 [1-Butyloctyl]benzene 0.41 MS/RI 1718 [1-pentylheptyl]benzene 0.33 MS 1795 [1-Methylundecyl]-benzene 0.40 MS Furans 855 2-Furanmethanol 8.59 MS/RI 947 Dihydro-5-methyl-2[3H]-furanone 2.91 MS/RI S-Containing compounds 903 Methional 0.49 MS/RI N-Containing compounds 752 1H-Pyrrole 1.51 MS/RI 990 2-Ethyl-5-methylpyrazine 0.56 MS/RI 1306 3-Butyl-2,5-dimethylpyrazine 0.37 MS/RI 1676 2,4-Di-t-butyl-6-nitro-phenol 0.85 MS Aliphatic and Terpene Hydrocarbons 1022 l-limonene 25.37 MS/RI 1094 Undecane 1.05 MS/RI 1190 Dodecane 0.94 MS/RI 1292 Tridecane 0.81 MS/RI 1391 Tetradecane 0.63 MS/RI 1689 Heptadecane 0.40 MS/RI 1782 1-Octadecene 0.62 MS/RI 1789 Octadecane 0.66 MS 1889 Nonadecane 0.50 MS/RI 1987 9-Methyl-nonadecane 0.34 MS Miscellaneous 986 7-Methyl-3-methylene-1,7-octadiene 11.47 MS/RI 1856 Bis[2-methylpropyl]ester-1,2-benzenedicarboxylic acid 0.39 MS 1952 Hexadecanoic acid 3.15 MS/RI - 87 -

RI A) Compounds P.A(%) ID B) Aldehydes and Ketones 1026 Benzaldehyde 33.34 MS/RI 1082 Nonanal 5.13 MS/RI 1085 Benzeneacetaldehyde 8.04 MS/RI 1038 1-Phenylethanone 9.07 MS 1173 Decanal 1.85 MS/RI 1278 3,4-Dimethyl-benzaldehyde 2.57 MS 1391 3,5-Dimethyl-2-octanone 1.22 MS 1554 5-Methyl-2-phenyl-2-hexenal 8.56 MS 1590 2-Methylundecanal 4.02 MS 1829 3,5-Di-tert-butyl-4-hydroxybenzaldehyde 5.37 MS N-Containing compounds 951 2,5-Dimethylpyrazine 0.48 MS/RI Aliphatic and Terpene Hydrocarbons 1249 Tetradecane 5.10 Miscellaneous compounds <700 Acetic acid 0.53 MS 1372 Methylundecanoate 2.16 MS/RI 1839 Hexadecanoic acid 10.74 MS/RI 2026 9-Octadecanoic acid 1.83 MS - 88 -

RI A) Possible Compounds Aroma description ID B) <700 unknown soy sauce SN 829 Furfural nutty, sweet MS/RI/SN 847 Isopropylbutanoate pungent RI/SN 867 2-Hexenal fatty, rancid RI/SN 855 2-Furfural almond, smoky, caramel-like MS/RI/SN 903 Methional cooked potato MS/RI/SN 921 Acetyl pyrroline nutty, roasted RI/SN 953 Benzaldehyde bitter almond, roasted MS/RI/SN 962 Dimethyl trisulfide pungent, sulfury MS/RI/SN 986 β-myrcene pungent, vinegar MS/RI/SN 999 l-limonene floral(lilac), grass MS/RI/SN 1035 Benzeneacetaldehyde sweet MS/RI/SN 1086 Dimethylethylpyrazine roasted RI/SN 1124 Methylcyclopentapyrazine roasted RI/SN 1230 Myrtenal spicy RI/SN 1302 2-Methoxy-4-vinylphenol spicy, nutty MS/RI/SN 1380 unknown spicy RI/SN 1452 α-humulene woody RI/SN 1465 Dodecyl alcohol oily MS/RI/SN 1481 5-Methyl-2-phenyl-2-hexenal roasted, cocoa MS/RI/SN 1490 Pentadecane alkane MS/RI/SN 1589 Hexadecane alkane MS/RI/SN 1645 β-caryophyllene alcohol earth RI/SN 1689 Heptadecane alkane MS/RI/SN 1714 unknown nutty SN 1738 unknown nutty SN 1782 1-Octadecene alkane MS/RI/SN - 89 -

RI A) Possible compound Aroma description ID B) 671 Thiophene garlic, pungent RI/SN 696 Methyl thiocyanate sulfury RI/SN 872 2-Furanmethanol oily, burnt MS/RI/SN 904 2,6-Dimethylpyrazine chocholate, roasted, nutty MS/RI/SN 914 Dimethyl sulfone smoky RI/SN 924 2-Cyclohexenone oily, pesticide MS/RI/SN 963 Benzaldehyde bitter almond, smoky, nutty MS/RI/SN 1084 Dimethylethylpyrazine nutty RI/SN 1102 unknown scorched rice, candy, nutty SN 1159 Diethylmethylpyrazine smoky, baked RI/SN 1181 (E,Z)-2,3-Nonadienal pungent, rubber RI/SN 1245 Tridecane alkane MS/RI/SN 1408 Methyl sulfide spicy, sulfury, garlic RI/SN 1757 unknown pungent, pine SN α α - 90 -

KOJI-AJI BS-100 Alanine 32.22 41.78 Glycine 74.84 128.31 α-aminobutylic acid 1.012 0 Valine 20.36 10.83 Leucine 53.96 26.28 Isoleucine 23.4 6.42 Threonine 31.76 8.86 Serine 52.96 12.31 Proline 198.3 61.67 Aspartic acid 27.87 20.21 Methionine 14.49 6.62 Glutamic acid 179.8 54.96 Phenylalanine 75.85 15.68 Ornithine 15.48 2.55 Lysine 8.086 31.95 Histidine 38.59 3.94 Tyrosine 28.3 1.01 Tryptophan 9.693 19.37 Cystine 3.144 3.46-91 -

(Relatve peak area) 200 150 100 50 ALA GLY ABA VAL LEU ILE THR SER PRO 0 ASP MET HYP GLU PHE ORN LYS HIS HLY TYR PHP TRP C-C KOJI-AJI BS-100 200000 150000 ASP GLU 100000 50000 0 0 50 100 150 200 ASP y = 1058.5x - 1240.8 R 2 = 0.9769 GLU y = 812.18x - 3366.8 R 2 = 0.9914 KOJI-AJI BS-100 aspartic acid 21.48 20.34 glutamic acid 142.22 55.03-92 -

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5'-CMP 5'-UMP 5'-GMP 5'-IMP KOJI-AJI 110.56 127.26 152.59 201.67 BS-100 0 6.21 164.92 198.83-95 -

Mohr 법표준용액결과 Ä ûá À 8 6 4 2 0 y = 17.59x - 0.015 R² = 0.999 계열 1 선형 ( 계열 1) -2 0 0.1 0.2 0.3 0.4 0.5 NaCl(g) 적정치 염도 KOJI-AJI 1.53 8.80% BS-100 3.77 21.49% - 96 -

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이미, 이취 부드러운맛강도 전체적인기호도 5.5 5 4.5 4 3.5 3 2.5 2 감칠맛강도 맛의지속성 대조군 RK 밀분해추출물 + Yeast Ext. 밀단백추출물 + 아로마일드 마늘양파의풍미 치킨풍미 - 98 -

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RI A) Possible compounds P.A(%) ID B) Alcohols 802 4-Methyl-3-hexanol 0.059 MS 1067 Thenylalcohol 1.554 MS 1107 Benzeneethanol 0.818 MS 1178 2-(2-Butoxyethoxy)-ethanol 0.858 MS 1495 2,6-Bis(1,1-dimethylethyl)-4-methyl-phenol 0.059 MS 2077 1-Octadecanol 0.058 MS S-containing compounds 863 2-Methyl-3-furanthiol 7.354 MS/RI 909 Methional 8.841 MS/RI 974 3-(Methylthio)-1-propanol 0.240 MS 986 Dihydro-2-methyl-3(2h)-thiophenone 0.788 MS 1013 2-Acetylthiazole 3.235 MS/RI 1111 5-Methyl-2-thiophenecarboxaldehyde 0.142 MS 1114 2-Formyl-5-methylthiophene 2.174 MS 1162 2-Methyl-3-(methyldithio)furan 1.147 MS 1273 4-Methyl-5-thiazoleethanol 17.199 MS 1405 2-Phenylthiophene 0.106 MS 1519 Bis(2-methyl-3-furyl)disulfide 8.245 MS N-containing compounds 1007 1H-Pyrrole-2-carboxaldehyde 0.214 MS 1059 1-(1H-Pyrrol-2-yl)-ethanone 0.979 MS 1091 N-(1,1-Dimethylethyl)-2-propenamide 0.252 MS - 103 -

1097 Butanal, dimethylhydrazone 0.248 MS 1127 N-(3-Methylbutyl)acetamide 0.087 MS 1201 Quinazoline 0.091 MS 1379 (Hydroxyimino)cyclohexane 1.340 MS Pyrazines 823 Methylpyrazine 0.439 MS/RI 996 Trimethylpyrazine 1.923 MS/RI 1071 3-Ethyl-2,5-dimethylpyrazine 0.207 MS 1079 Tetramethylpyrazine 0.681 MS/RI 1304 2,5-Dimethyl-3-n-butylpyrazine 0.176 MS 1377 1,4-Dimethylpyrrolo(1,2-a)pyrazine 0.115 MS 1570 2,5-Dimethyl-3-n-butylpyrazine 0.145 MS Furans 851 2-Furanmethanol 0.460 MS/RI 949 2-Furancarboxylic acid 0.932 MS Acids 1468 4-Hydroxy-3,5-dimethoxy-benzoic acid 0.074 MS 1955 n-hexadecanoic acid 1.478 MS/RI >2100 9,12-Octadecadienoic acid 0.212 MS/RI >2100 (z)-9-octadecenoic acid 0.104 MS/RI Aldehyde and Ketone 958 Benzaldehyde 0.716 MS/RI 1146 4-Heptanone 0.079 MS 1185 4-Fluoro-3-hydroxybenzaldehyde 0.228 MS 1260 α-ethylidenbenzeneacetaldehyde 0.092 MS 1386 3-Methoxy-4-hydroxybenzaldehyde 0.227 MS Aliphatic hydrocarbons 831 1,2-Nonadiene 3.362 MS 1024 Dotriacontane 0.065 MS 1192 Dodecane 0.072 MS/RI 1393 Tetradecane 0.108 MS/RI 1592 Hexadecane 0.180 MS/RI 1655 2-Methylhexadecane 0.070 MS 1691 Heptadecane 0.436 MS/RI 1758 2-Methylheptadecane 0.187 MS 1796 Octadecane 0.542 MS/RI 1898 Nonadecane 0.233 MS/RI Ester 1197 β-nonalactone 0.070 MS 1341 2,2,4-Trimethyl-1,3-pentanediol diisobutyrate 1.548 MS 1365 Butyl butyrate 1.287 MS 1987 Ethyl hexadecanoate 0.753 MS/RI >2100 Ethyl linoleate 0.880 MS - 104 -

RI A) Possible compounds P.A(%) ID B) Alcohols 1178 2-(2-Butoxyethoxy)-ethanol 0.258 MS S-containing compounds 864 2-Methyl-3-furanthiol 10.157 MS/RI 910 Methional 6.897 MS/RI 949 2,3-Dimethylthiophene 1.666 MS 986 Dihydro-2-methyl-3(2h)-thiophenone 1.918 MS 1013 2-Acetylthiazole 2.689 MS/RI 1068 2-Thiophenemethanol 2.467 MS 1091 5-Acetylthiazole 0.220 MS 1098 3-Thienylmethanol 0.141 MS 1114 2-Formyl-5-methylthiophene 1.344 MS 1162 2-Methyl-3-(methyldithio)furan 1.066 MS 1164 1,2-Dithian-4-one 0.460 MS 1224 2-Isopropyl-4,5-dimethylthiazole 0.278 MS 1230 2,5-Thiophenedicarboxaldehyde 0.572 MS 1234 2-Methyl-3-(methylthio)furan 0.142 MS 1278 4-Methyl-5-thiazoleethanol 25.051 MS 1405 3-Phenyl-thiophene 0.151 MS 1518 Bis(2-methyl-3-furyl)disulfide 5.953 MS 1610 1-Isothiocyanato-heptane 0.241 MS 1712 2-Methyl-3-furanthiol 0.713 MS N-containing compounds 1535 3-Methyl-quinoline 0.112 MS 1567 2-Ethyl-quinoline 0.078 MS Pyrazines 822 Methylpyrazine 0.389 MS/RI 996 Trimethylpyrazine 3.630 MS/RI 1078 3-Ethyl-2,5-dimethylpyrazine 0.640 MS 1189 2-(2-Methylpropyl)-3,6-dimethylpyrazine 0.087 MS 1294 2,5-Dimethyl-3-(3-methylbutyl)pyrazine 0.852 MS 1318 2-Butyl-3,5-dimethylpyrazine 0.088 MS 1570 2,5-Dimethyl-3-n-butylpyrazine 0.205 MS Furans 851 2-Furanmethanol 0.157 MS/RI Acids - 105 -

1957 Hexadecanoic acid 1.428 MS/RI >2100 9-Octadecenoic acid 0.119 MS Aldehyde and Ketone 958 Benzaldehyde 0.789 MS/RI 1041 Benzeneacetaldehyde 8.469 MS 1099 Nonanal 0.163 MS/RI 1748 3,5-Di-tert-butyl-4-hydroxybenzaldehyde 0.159 MS Ester 1365 3-Hydroxy-2,4,4-trimethylpentyl 2-methylpropanoate 0.948 MS 1988 Ethyl hexadecanoate 1.048 MS/RI 2016 Isopropyl palmitate 0.106 MS >2100 Ethyl linoleate 1.329 MS Aliphatic hydrocarbons 973 1-Ethyl-2-methyl-benzene 0.218 MS 1094 Undecane 0.138 MS/RI 1592 Hexadecane 0.144 MS/RI 1691 Heptadecane 0.237 MS/RI 1766 3-Methyl-heptadecane 0.111 MS 1796 Octadecane 0.352 MS/RI 1878 Cyclohexadecane 0.136 MS/RI 1899 Nonadecane 1.116 MS/RI 2077 1-Octadecene 0.116 MS Miscellaneous compounds 1104 Maltol 1.441 MS 1138 2,3-Dihydro-3,5-dihydroxy-6-methyl-4h-pyran-4-one 1.219 MS 1185 Methylphloroglucinol 0.547 MS 1341 2-Methyl-propanoic acid 0.435 MS RI A) Possible compounds P.A(%) ID B) Alcohols 1024 2-Ethyl-1-hexanol 0.208 MS 1177 2-(2-Butoxyethoxy)-ethanol 0.378 MS S-containing compounds 865 2-Methyl-3-furanthiol 25.577 MS/RI 908 Methional 8.708 MS/RI 986 Dihydro-2-methyl-3(2H)-thiophenone 0.746 MS 1012 2-Acetylthiazole 1.661 MS/RI 1065 2-Thienylmethanol 1.887 MS - 106 -

1097 3-Thienylmethanol 0.1 MS 1110 2-Formyl-5-methylthiophene 0.237 MS 1113 2-Formyl-3-methylthiophene 1.974 MS 1161 2-Methyl-3-(methyldithio)furan 0.96 MS 1169 2-Butyl-thiophene 0.117 MS 1230 Dimethylformylthiophene 0.418 MS 1271 4-Methyl-5-thiazoleethanol 14.289 MS 1405 3-Phenyl-thiophene 0.189 MS 1517 Bis(2-methyl-3-furyl)disulfide 2.141 MS N-containing compounds 992 N,N-Dimethyl-4-pyridinamine 0.093 MS Pyrazines 824 Methylpyrazine 0.523 MS/RI 995 Trimethylpyrazine 1.64 MS 1072 2-Ethyl-3,5-dimethylpyrazine 0.227 MS 1185 1-Methyl-3,4-dihydro-pyrrolo(1,2-a)pyrazine 0.15 MS 1293 2-Butyl-3,5-dimethyl-pyrazine 0.213 MS Furans 852 2-Furanmethanol 0.77 MS/RI 948 2-Furancarboxylic acid 1.171 MS Acids 1951 Hexadecanoic acid 0.082 MS/RI Aldehyde 958 Benzaldehyde 0.738 MS/RI Ester 1341 2,2,4-Trimethylpentanediol-1,3-diisobutyrate 0.557 MS 1987 Ethyl hexadecanoate 0.236 MS/RI >2000 Ethyl linoleate 0.179 MS >2000 Ethyl oleate 0.142 MS Aliphatic hydrocarbons 830 1,2-Nonadiene 3.537 MS 858 Ethylbenzene 2.343 MS/RI 1095 Undecane 0.134 MS/RI 1592 Hexadecane 0.121 MS/RI 1691 Heptadecane 0.343 MS/RI 1765 3-Methyl-heptadecane 0.117 MS 1796 Octadecane 0.326 MS/RI 1898 Nonadecane 0.35 MS/RI Miscellaneous compounds 803 2,2-Dimethyl-3-pentanol 0.136 MS 890 1,4-Dimethylbenzene 1.316 MS/RI 973 2-Acetylcyclopentanone 0.254 MS 1135 2,3-Dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one 0.945 MS - 107 -

RI A) Possible compounds P.A(%) ID B) Alcohols 1178 2-(2-Butoxyethoxy)-ethanol 0.669 MS 1213 2-Naphthol 0.056 MS 1757 2-(Octadecyloxy)-ethanol 0.206 MS S-containing compounds 864 2-Methyl-3-furanthiol 6.745 MS/RI 910 Methional 8.84 MS/RI 986 Dihydro-2-methyl-3(2H)-thiophenone 0.698 MS 1013 2-Acetylthiazole 2.202 MS/RI 1116 2-Formyl-3-methylthiophene 5.897 MS 1162 2-Methyl-3-(methyldithio)furan 1.511 MS 1165 1,2-Dithian-4-one 0.203 MS 1190 3-Ethyl-2-formylthiophene 0.161 MS 1230 2,5-Diformylthiophene 0.609 MS 1270 4-Methyl-5-thiazoleethanol 4.527 MS 1405 3-Phenyl-thiophene 0.056 MS 1519 Bis(2-methyl-3-furyl)disulfide 9.639 MS N-containing compounds 1026 1-(2-Pyridinyl)-ethanone 0.341 MS 1068 1-(1H-Pyrrol-2-yl)-ethanone 6.318 MS 1080 1,7-Dihydro-6H-purin-6-one 0.844 MS 1089 Butanal, ethylhydrazone 0.063 MS 1201 Quinazoline 0.125 MS 1293 2-Methyl-quinoxaline 0.475 MS 1411 1-(3-Methylphenyl)-1H-imidazole 0.032 MS 1450 N-(4-Chlorophenyl)-acetamide 2.688 MS 1874 N-Ethylpiperazine 0.038 MS Pyrazines 823 Methylpyrazine 0.179 MS/RI 996 Trimethylpyrazine 2.394 MS/RI 1305 2,5-Dimethyl-3-(3-methylbutyl)-pyrazine 0.206 MS 1570 2-Butyl-3,5-dimethylpyrazine 0.117 MS Furans 852 2-Furanmethanol 1.106 MS/RI 949 2-Furancarboxylic acid 1.337 MS - 108 -

Acids 1357 Octadecanoic acid 0.036 MS 1552 Dodecanoic acid 0.093 MS/RI 1957 Hexadecanoic acid 1.92 MS/RI >2100 9,12-Octadecadienoic acid 0.191 MS/RI >2100 9-Octadecenoic acid 0.245 MS/RI Aldehyde and Ketone 959 Benzaldehyde 0.589 MS/RI 1041 Benzeneacetaldehyde 10.76 MS 1100 Nonanal 0.209 MS/RI 1197 Decanal 0.077 MS/RI 1387 4-Hydroxy-3-methoxy-benzaldehyde 0.129 MS 1401 Dodecanal 0.053 MS/RI 1476 5-Methyl-2-phenyl-2-hexenal 0.132 MS/RI 2007 2,2-Dimethyl-3-octanone 0.145 MS Ester 1341 2,2,4-Trimethylpentanediol-1,3-diisobutyrate 0.998 MS 1365 Hexyl isobutanoate 1.349 MS 1988 Ethyl palmitate 0.673 MS/RI >2100 Ethyl linoleate 1.121 MS Aliphatic hydrocarbons 1095 Undecane 0.23 MS/RI 1192 Dodecane 0.101 MS/RI 1393 Tetradecane 0.131 MS/RI 1467 (E)-2-Tetradecene 0.082 MS/RI 1592 Hexadecane 0.168 MS/RI 1655 2-Methyl-hexadecane 0.056 MS 1691 Heptadecane 0.411 MS/RI 1723 2,3-Dimethyl-bicyclo(2.2.1)hept-2-ene 0.278 MS 1796 Octadecane 0.517 MS/RI 1898 Nonadecane 0.236 MS/RI 2077 1-Nonadecene 0.045 MS Miscellaneous compounds 890 1,4-Dimethylbenzene 0.094 MS/RI 1106 Maltol 2.086 MS 1138 2,3-Dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one 1.14 MS 1302 2-Methoxy-4-vinylphenol 0.208 MS/RI 1398 2,3,6-Trimethyl-phenol 0.082 MS - 109 -

RI A) Possible compounds P.A(%) ID B) Alcohols 1024 1-Dodecanol 0.137 MS 1106 Benzeneethanol 0.791 MS 1120 3,7,11,15-Tetramethyl-2-hexadecen-1-ol 0.099 MS 1177 2-(2-Butoxyethoxy)-ethanol 0.708 MS S-containing compounds 864 2-Methyl-3-furanthiol 7.051 MS/RI 909 Methional 8.238 MS/RI 965 Dimethyl trisulfide 0.059 MS 974 3-(Methylthio)-1-propanol 0.227 MS 986 Dihydro-2-methyl-3(2H)-thiophenone 0.48 MS 1013 2-Acetylthiazole 2.995 MS/RI 1065 2-Thiophenemethanol 1.992 MS 1083 2-Ethyl-5-methylthiophene 0.041 MS 1097 3-Thiophenemethanol 0.173 MS 1111 2-Formyl-5-methylthiophene 0.363 MS 1114 2-Formyl-3-methylthiophene 4.316 MS 1162 2-Methyl-3-(methyldithio)furan 1.11 MS 1189 3-Ethyl-2-formylthiophene 0.095 MS 1269 4-Methyl-5-thiazoleethanol 8.726 MS 1405 1-Naphthalenethiol 0.064 MS 1517 Bis(2-methyl-3-furyl)disulfide 3.777 MS N-containing compounds 843 3-Methyl-1H-pyrrole 0.07 MS 1060 1-(1H-Pyrrol-2-yl)-ethanone 2.867 MS Pyrazines 825 Methylpyrazine 0.682 MS/RI 993 2-Ethyl-6-methyl-pyrazine 0.261 MS 996 Trimethylpyrazine 1.571 MS/RI 1071 2-Ethyl-3,5-dimethyl-Pyrazine 0.228 MS 1304 2,5-Dimethyl-3-(3-methylbutyl)-pyrazine 0.08 MS 1376 1,4-Dimethylpyrrolo(1,2-a)pyrazine 0.092 MS Furans 810 Dihydro-2-methyl-3(2H)-furanone 0.12 MS 852 2-Furanmethanol 0.508 MS/RI 949 2-Furancarboxylic acid 1.64 MS 1151 2-Methyl-3-(methylthio)furan 0.06 MS 1213 3-Phenylfuran 0.08 MS/RI Acids 1549 Dodecanoic acid 0.077 MS/RI - 110 -

1954 Hexadecanoic acid 1.618 MS/RI >2100 9,12-Octadecadienoic acid 0.111 MS/RI Aldehyde and Ketone 929 2-Cyclohexen-1-one 0.14 MS/RI 958 Benzaldehyde 0.685 MS/RI 1006 1H-Pyrrole-2-carboxaldehyde 0.207 MS 1035 2-Fluoro-benzaldehyde 0.063 MS 1040 Benzeneacetaldehyde 11.558 MS 1185 4-Fluoro-3-hydroxy-benzaldehyde 0.245 MS 1230 2,5-Thiophenedicarboxaldehyde 0.628 MS Ester 803 Methyl 3-butenoate 0.089 MS 1341 2,2,4-Trimethylpentanediol-1,3-diisobutyrate 1.354 MS 1365 Butyl butanoate 1.442 MS 1987 Ethyl palmitate 1.063 MS/RI >2100 Ethyl linoleate 1.484 MS Aliphatic hydrocarbons 1095 Undecane 0.136 MS/RI 1192 Dodecane 0.085 MS/RI 1393 Tetradecane 0.072 MS/RI 1556 2-Methylpentadecane 0.066 MS 1592 Hexadecane 0.261 MS/RI 1655 2-Methylhexadecane 0.142 MS/RI 1691 Heptadecane 0.76 MS/RI 1757 2-Methylheptadecane 0.318 MS/RI 1766 3-Methylheptadecane 0.288 MS/RI 1796 Octadecane 1.011 MS/RI 1898 Nonadecane 1.17 MS/RI 2077 1-Nonadecene 0.069 MS Miscellaneous compounds 944 Chlorobenzene 0.099 MS 1134 2,3-Dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one 0.086 MS 1145 (z)-sabinene hydrate 0.061 MS 1299 2,6-Dimethylfluorobenzene 0.037 MS 1301 2-Methoxy-4-vinylphenol 0.478 MS/RI 1331 2-Chloroethoxybenzene 0.079 MS 1453 2,6-Bis(1,1-dimethylethyl)-2,5-cyclohexadiene-1,4-dione 0.064 MS 1494 2,6-Bis(1,1-dimethylethyl)-4-methylphenol 0.281 MS 1497 2,4-Bis(1,1-dimethylethyl)-phenol 0.521 MS 1502 Tributylphsophate 0.097 MS - 111 -

RI A) Possible Compounds Aroma description ID B) 846 Methylpyrazine nutty MS/RI/SN 872 unknown sulfury SN 917 Methional cooked potato MS/RI/SN 936 Benzaldehyde bitter almond, roasted MS/RI/SN 1088 Tetramethylpyrazine peanuts MS/RI/SN 1104 Benzeneethanol rose-like+sweet MS/RI/SN 1138 5-Methyl-2-thiophenecarboxaldehyde roasted peanuts MS/RI/SN 1220 3-Pyridinecarboxylic acid, methyl ester nutty MS/RI/SN 1267 α-ethylidenebenzeneacetaldehyde potato chips MS/RI/SN 1295 unknown smoky+nutty SN 1305 unknown roasted nuts SN 1362 unknown vinegar SN 1381 4-Hydroxy-3-methoxybenzaldehyde sweet MS/RI/SN 1693 Heptadecane alkane MS/RI/SN 1820 Octadecane alkane MS/RI/SN 1979 unknown nutty SN 1999 unknown nutty SN - 112 -

- 113 -

α - 114 -

- 115 -

- 116 -

0.14 0.12 0.1 0.08 0.06 0.04 0.02 0 1 2 3 AVR GLU 전 GLU 후 μ α - 117 -

- 118 -

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μ μ - 143 -

- 144 -

α α - 145 -

α - 146 -

μ - 147 -

α α μ μ μ μ μ - 148 -

μ μ μ - 149 -

- 150 -

γ γ γ - 151 -

시약 장단점 - 152 -

- 153 -

- 154 -

Absorbance at 305nm 0.5 0.4 0.3 0.2 0.1 y = 0.0021x + 0.0164 R 2 = 0.9914 0.0 0 50 100 150 200 Glutathione concentration (um) - 155 -

25,000,000 20,000,000 y = 4E+06x + 520070 R 2 = 0.9977 Area 15,000,000 10,000,000 5,000,000 0 0 1 2 3 4 5 Glutathione concentration (mm) 5 mm 4 mm 3 mm 2 mm 1 mm - 156 -

1,000 12% T, 1% C 15% T, 1% C 16.5% T, 3% C 20% T, 2.6% C Molecular mass (kda) 100 10 1 0.0 0.2 0.4 0.6 0.8 1.0 Relative mobility Protein (kda) 116 97 66 55 45 36 1 2 3 4 5 6 Protein (kda) 26.6 17.0 14.2 6.5 3.5-157 -

- 158 -

mau 2000 AJI mau 600 BS-100 1400 500 400 1000 300 600 200 200 0 0 20 40 60 80 Time (min) 100 0 0 20 40 60 80 Time (min) - 159 -

- 160 -

- 161 -

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- 163 -

Sample 1) WP A1 A2 A4 A8 A12 A24 F1 F2 F4 F8 F12 F24 P1 P2 P4 P8 P12 P24 Asp 0.02 0.02 0.03 0.03 0.03 0.04 0.00 0.06 0.08 0.10 0.09 0.04 0.00 0.00 0.03 0.00 0.03 0.00 0.00 Thr 0.09 0.22 0.27 0.35 0.52 0.58 0.63 11.53 2.07 2.77 3.50 3.69 3.20 0.12 0.26 0.37 0.55 0.68 0.48 Ser 0.02 0.08 0.10 0.14 0.19 0.22 0.08 0.33 0.49 0.64 0.89 0.86 0.38 0.01 0.10 0.16 0.23 0.24 0.10 Glu 0.21 0.19 0.20 0.20 0.24 0.25 0.41 0.22 0.26 0.29 0.30 0.32 0.56 0.00 0.20 0.21 0.17 0.15 0.24 Pro 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.10 0.53 0.00 0.00 0.00 0.00 0.00 0.00 Gly 0.00 0.00 0.01 0.00 0.00 0.02 0.00 0.10 0.09 0.13 0.23 0.25 0.39 0.00 0.00 0.00 0.00 0.03 0.08 Ala 0.00 0.00 0.00 0.07 0.11 0.13 0.17 0.12 0.17 0.25 0.34 0.39 1.16 0.00 0.00 0.02 0.02 0.18 0.66 Cys 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.03 0.04 0.09 0.00 0.00 0.20 0.34 0.00 0.00 Val 0.25 0.31 0.17 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.48 0.61 0.88 0.00 0.00 0.00 0.00 0.00 0.00 Met 0.04 0.07 0.09 0.10 0.13 0.15 0.18 0.16 0.20 0.26 0.34 0.35 0.31 0.08 0.10 0.12 0.16 0.17 0.22 Ile 0.01 0.03 0.00 0.04 0.07 0.08 0.10 0.20 0.35 0.49 0.64 0.69 0.85 0.04 0.06 0.10 0.17 0.22 0.39 Leu 0.12 0.18 0.20 0.23 0.32 0.35 0.59 0.70 0.99 1.25 1.49 1.53 1.82 0.19 0.23 0.31 0.46 0.59 1.14 Tyr 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.45 0.49 0.71 0.00 0.00 0.00 0.00 0.05 0.08 Phe 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.42 0.51 0.95 0.00 0.00 0.00 0.00 0.00 0.45 His 0.61 0.52 0.67 0.62 0.57 0.71 0.63 0.59 0.62 0.59 0.60 0.64 0.57 0.69 0.70 0.67 0.65 0.65 0.63 Lys 0.01 0.03 0.04 0.05 0.08 0.09 0.08 0.18 0.22 0.26 0.29 0.30 0.25 0.00 0.00 0.00 0.00 0.11 0.10 NH 3 0.25 0.25 0.34 0.28 0.31 0.34 0.53 0.31 0.31 0.34 0.34 0.39 1.05 0.23 0.25 0.25 0.27 0.33 0.67 Arg 0.00 0.08 0.00 0.14 0.17 0.15 0.00 0.41 0.48 0.53 0.60 0.53 0.13 0.12 0.12 0.15 0.16 0.00 0.09 Total (%) 1.63 1.98 2.12 2.25 2.74 3.11 3.40 4.90 6.37 7.95 11.03 11.73 13.83 1.48 2.05 2.56 3.21 3.40 5.33-164 -

( 다 ) 효모변이에따른 GSH 추출물의 DPPH 라디컬소거능 - 165 -

(2) 가수분해시간에따른펩타이드패턴과고꾸미및생리활성탐색 ( 가 ) 효소종류 / 가수분해도에따른펩타이드패턴분석 1 효소종류 / 가수분해도에따른시료제조 2 Reversed phase HPLC chromatography - 166 -

- 167 -

- 168 -

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- 171 -

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- 173 -

다양한산소유래라디컬중 hydroxyl 라디컬은단백질, DNA, PUFA, nucleic acid와같은생체분자를공격하는유해성을보이므로 hydroxyl 라디컬을제거함으로써관련질병에대항하여인체를보호할수있다. 각각의가수분해물의 hydroxyl 라디컬소거능은 0.5-4.0 mg/ml 범위에서농도의존적인활성증가를보였으나, 효소종류 / 가수분해시간에따른활성차이는보이지않았다 (Fig. 73). 특히, 우수한고꾸미를보인 A24h의 hydroxyl radical 소거능의 IC50은 0.39 mg/ml로 alcalase에의해가수분해된 wheat germ 가수분해물 (IC50 0.12 mg/ml) 보다약 2배정도낮은결과를나타낸반면, chickpea 단백가수분해물 (IC50 약 1.5 mg/ml) 에비해서는약 4배높은결과를나타내었다. - 174 -

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μ - 180 -

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Comparison of volatile Maillard reaction products from tagatose and other reducing sugars with amino acids, In Hee Cho, Sarah Lee, Hae-Roung Jun, Hoe-Jin Roh, and Young-Suk Ki, Food Sci. Biotechnol. 19(2): 431-438 (2010) Preparation and characterization of taste-active compounds by enzymatic hydrolysis of wheat gluten, Food Research International ( 투고준비중, 2010년 5월 ) Umami Evaluation and Biological Activities of Wheat Protein Hydrolysates with Different Degree of Hydrolysis,14th World Congress of Food Science &Technology, (TS13-41) Oct 19-23, 2008, Sanghai, China (session time: Oct 21, Tuesday, Morning) Kokumi evaluation and biological activities of domestic and imported wheat protein hydrolysates produced by different proteases,2009 IFT Annual Meeting, (058-13) Jun 6-9, Anaheim, CA, USA (Session Time: Sunday Jun 7, 2009 3:00 PM - 5:00 PM) 미국식품과학회 (Institute of Food Technologists) - 192 -

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제 7 장 참고문헌 - 194 -

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