中国农业科学 ›› 2020, Vol. 53 ›› Issue (8): 1627-1642.doi: 10.3864/j.issn.0578-1752.2020.08.012
赵文华1,2,王桂瑛1,2,荀文1,2,俞媛瑞1,2,葛长荣2(),廖国周2(
)
收稿日期:
2019-10-17
接受日期:
2020-02-01
出版日期:
2020-04-16
发布日期:
2020-04-29
通讯作者:
葛长荣,廖国周
作者简介:
赵文华,E-mail: 598363924@qq.com。
基金资助:
ZHAO WenHua1,2,WANG GuiYing1,2,XUN Wen1,2,YU YuanRui1,2,GE ChangRong2(),LIAO GuoZhou2(
)
Received:
2019-10-17
Accepted:
2020-02-01
Online:
2020-04-16
Published:
2020-04-29
Contact:
ChangRong GE,GuoZhou LIAO
摘要:
【目的】利用代谢组学结合多元统计学方法筛选热处理前后茶花鸡胸肌和腿肌中的生物标志物,旨在明确表征茶花鸡不同部位肌肉中特征风味的水溶性化合物,为建立优质地方鸡评价标准提供科学理论依据。【方法】以300日龄的云南茶花鸡为研究对象,采用液相色谱-四极杆静电场轨道阱-质谱法(LC-Q-Exactive-MS)检测热处理前后胸肌和腿肌中的水溶性化合物,基于数据库Metlin进行全谱鉴定,以2-氯苯丙氨酸浓度计算各物质相对含量,并对检测出的物质进行分析,利用正交偏最小二乘法(orthogonal partial least squares discrimination analysis,OPLS-DA)建立代谢物表达量与样本组之间的关系模型,来实现对样品类别的预测,通过S-plot图初步筛选具有代表性的生物标志物,观察置换200次检验图的直线斜率判定模型是否存在过拟合现象,并且通过计算变量投影重要度(variable important for the Projection,VIP)判别各代谢物表达模式下对各组样本分类判别的影响强度和解释能力,达到辅助筛选标志物的目的;通过相对含量结果计算滋味活性值(taste activity value,TAV),结合感官评定结果评价茶花鸡肌肉的特征风味。【结果】茶花鸡肌肉在质谱正离子和负离子模式下,分别检测出106种和43种生物代谢物,其中主要成分包括氨基酸类、维生素类、核苷类、有机酸类、脂肪酸类等,其中肌酸、肌肽、氨基酸和核苷酸为茶花鸡肌肉热处理前后存在的差异性水溶性化合物,可作为特征滋味贡献物质的生物标志物;TAV值结果中显示,胸肌中TAV值最大的是γ-氨基丁酸(TAV=23.00),其次是组氨酸(TAV=1.14);腿肌中TAV值最大的是γ-氨基丁酸(TAV=37.50),其次是甜菜碱(TAV=2.00);通过感官评定发现胸肌和腿肌整体滋味以甜鲜味为主。【结论】利用代谢组学分析得出茶花鸡特征滋味物质,并结合滋味理论值和感官评定值确定茶花鸡胸肌和腿肌的整体滋味以甜鲜肉香味为主,且腿肌优于胸肌。
赵文华,王桂瑛,荀文,俞媛瑞,葛长荣,廖国周. 基于代谢组学筛选表征茶花鸡肌肉中特征风味的水溶性化合物[J]. 中国农业科学, 2020, 53(8): 1627-1642.
ZHAO WenHua,WANG GuiYing,XUN Wen,YU YuanRui,GE ChangRong,LIAO GuoZhou. Selection of Water-Soluble Compounds by Characteristic Flavor in Chahua Chicken Muscles Based on Metabolomics[J]. Scientia Agricultura Sinica, 2020, 53(8): 1627-1642.
表2
正离子模式下肌肉样本代谢产物"
编号 No. | 代谢物 Metabolite | 编号 No. | 代谢物 Metabolite | 编号 No. | 代谢物 Metabolite |
---|---|---|---|---|---|
1 | 肌酸 Creatine | 501 | 氨基丁酸 gamma-Aminobutyric acid | 1156 | 黄嘌呤 Xanthine |
3 | L-鹅肌肽 L-Anserine | 517 | 泛酸 Pantothenic Acid | 1213 | α-亚麻酸 α-Linolenic Acid |
11 | 肌酐 Creatinine | 525 | L-异亮氨酸 L-Isoleucine | 1236 | 高香草基酸Homovanillic acid |
13 | 左旋肉碱 L-Carnitine | 546 | 磷酸胆碱Phosphocholine | 1259 | 鸟嘌呤 Guanine |
15 | 甜菜碱 Betaine | 553 | 5'-甲基硫腺苷 5'-Methylthioadenosine | 1296 | 聚氧乙烯辛烷基苯酚醚 9-OxoODE |
37 | 乙酰肉碱 Acetylcarnitine | 557 | 鞘氨醇 Sphingosine | 1308 | 烟酰胺 Niacinamide |
44 | N-[4-(乙酰基氨基)苯基]-3-氧代丁酰胺 Hydrouracil | 561 | 叔亮氨酸 Butyrylcarnitine | 1363 | 二氢鞘氨醇 Sphinganine |
65 | 亚油酸肉碱 Linoleylcarnitine | 563 | 甘油磷酸胆碱 Glycerophosphocholine | 1403 | 1-肉豆蔻酰-2-羟基-sn-甘油-3-磷酸胆碱LysoPC(14:0) |
88 | 1-甲基组氨酸 1-Methylhistidine | 579 | L-精氨酸 L-Arginine | 1431 | 丙氨酰 - 脯氨酸 Alanyl-Proline |
108 | 次黄嘌呤 Hypoxanthine | 591 | 1-十六烷酰-sn-甘油-3-磷酸乙醇胺 LysoPE (0:0/16:0) | 1446 | 鸟嘌呤核苷 Guanosine |
124 | L-氯化棕榈酰肉碱 L-Palmitoylcarnitine | 617 | 二十四碳六烯酸 tetracosahexaenoic acid | 1458 | 苯乙酸 Phenylacetic acid |
146 | L-苯丙氨酸 L-Phenylalanine | 618 | L-丝氨酸 L-Serine | 1472 | 硬脂酸酰胺 Stearamide |
148 | 1-十五烷氧基-2-羟基-sn-甘油-3-磷酸胆碱 LysoPC(15:0) | 625 | N-十四碳酰-L-丝氨酸 Decanoylcarnitine | 1513 | 腺苷酸 Adenosine monophosphate |
219 | 异亮脯氨酸 Isoleucylproline | 641 | 牛磺胆酸 Taurocholic acid | 1518 | 油酸 Oleic Acid |
228 | L-组氨酸 L-Histidine | 676 | 高肌肽 Homocarnosine | 1558 | N6-乙酰-L-赖氨酸 N6-Acetyl-L-lysine |
233 | 乙酰胆碱 Acetylcholine | 692 | 植物鞘氨醇 Phytosphingosine | 1594 | α-维生素E α-Tocopherol |
237 | N-乙酰-L-组氨酸 N-Acetyl-L-Histidine | 694 | DL-2-氨基正辛酸 DL-2-Aminooctanoic acid | 1607 | 亚油酸 Linoleic acid |
242 | 肌苷 Inosine | 712 | 苯丙酮酸 Phenylpyruvic acid | 1608 | 维生素 ARetinol |
247 | 胆碱 Choline | 733 | L-脯氨酰基-L-苯丙氨酸L-prolyl-L-phenylalanine | 1611 | 腺嘌呤Adenine |
255 | 3-(硬脂酰氧基)-4-(三甲基铵基)丁酸酯 DL-Stearoylcarnitine | 753 | 顺-13-二十二烯酰胺 13Z-Docosenamide | 1633 | 油酸酰胺 Oleamide |
282 | L-谷氨酰胺 L-Glutamine | 809 | 腺苷 Adenosine | 1717 | 1-磷酸鞘氨醇 Sphingosine 1-phosphate |
283 | 1-苯乙胺 1-Phenylethylamine | 829 | 2-苯基乙酰胺 2-Phenylacetamide | 1861 | 尿嘧啶 Uracil |
288 | 谷胱甘肽 Glutathione | 835 | 花生酸 Arachidonic Acid | 1877 | 谷酰氨基-色氨酸 Glutaminyl-Tryptophan |
302 | 十四左旋肉碱 Tetradecanoylcarnitine | 897 | L-谷氨酸 L-Glutamate | 1929 | 烟酸 Niacin |
303 | L-色氨酸 L-Tryptophan | 922 | 二十二酰胺 Docosanamide | 1944 | 十八酰胺乙醇 Stearoylethanolamide |
332 | 1-棕榈酰-2-羟基-sn-甘油-3-磷酸胆碱 LysoPC(16:0) | 927 | 组织胺 Histamine | 1991 | 脱氧胞苷 Deoxycytidine |
357 | β-丙氨酸 β-Alanine | 936 | 肌苷酸 IMP | 1994 | 苹果酸 Malic acid |
386 | 磺熊去氧胆酸 Tauroursodeoxycholic acid | 959 | L-2-氨乙基丝氨酸磷酸酯 L-2-Aminoethyl seryl phosphate | 2099 | 脱氧腺嘌呤核苷 Deoxyadenosine |
414 | 辛酰基肉碱 L-Octanoylcarnitine | 1005 | L-苏氨酸 L-Threonine | 2103 | 甘氨酰-L-亮氨酸 Glycyl-L-leucine |
427 | 焦谷氨酸 Pyroglutamic acid | 1017 | 吡哆胺 Pyridoxamine | 2104 | 尿刊酸 Urocanic acid |
446 | L-酪氨酸 L-Tyrosine | 1056 | 2-哌啶甲酸 DL-pipecolic acid | 2138 | 脯氨酸甜菜碱 Prolinebetaine |
459 | 牛磺酸 Taurine | 1067 | 乙酰双氢硫辛酸酰胺 S-Acetyldihydrolipoamide | 2143 | 尿酸 Uric acid |
465 | 棕榈酸酰胺 Palmitic amide | 1078 | 邻苯二甲酸二丁酯 Alpha-CEHC | 2203 | 亚油酰胺 Linoleamide |
467 | L-脯氨酸 L-Proline | 1092 | 3Α-羟基-7-氧代-5Β-胆烷酸 Nutriacholic acid | 2291 | 维生素A酸 Retinoic acid |
476 | 磷酸羟基丙酮酸Phosphohydroxypyruvic acid | 1114 | 蛋氨酸亚矾 Methionine sulfoxide | 2296 | 苯甲酸 Benzoic acid |
2036 | 5-L-谷氨酰-L-半胱氨酸 γ-Glu-Cys |
表3
负离子模式下肌肉样本代谢产物"
编号 No. | 代谢物 Metabolite | 编号 No. | 代谢物 Metabolite |
---|---|---|---|
356 | 尿苷 Uridine | 1102 | L-谷氨酰基5-磷酸 L-Glutamyl 5-phosphate |
499 | 十一烷二酸 Undecanedioic acid | 1083 | 吲哚乳酸 Indolelactic acid |
1762 | 血栓素 B3TXB3 | 1793 | 谷氨酰二肽 Glutamylphenylalanine |
258 | 苏糖酸 Threonic acid | 1019 | 谷氨酰丙氨酸 Glutamylalanine |
1454 | 四氢皮质醇 Tetrahydrocortisol | 318 | γ-谷氨酰谷氨酸 Gamma Glutamylglutamic acid |
533 | 四氢皮质甾酮 Tetrahydrocorticosterone | 1778 | 二十烷二酸 Eicosanedioic acid |
1323 | 硬脂肪酸 Stearic acid | 1343 | 脱氧尿苷一磷酸 dUMP |
875 | 鞘氨醇1-磷酸 Sphingosine 1-phosphate | 1814 | 十二烷二酸 Dodecanedioic acid |
139 | 景天庚酮糖 Sedoheptulose | 198 | D-葡萄糖-6-磷酸二钠 D-Glucose 6-phosphate |
276 | 脯氨酰羟脯氨酸 Prolylhydroxyproline | 482 | 脱氧胆酸 Deoxycholic acid |
1658 | 苯乙酮酸 Phenylglyoxylic acid | 1609 | D-生物素 D-Biotin |
1756 | 苯丁酰谷氨酰胺 Phenylbutyrylglutamine | 1811 | 皮质醇 Cortisol |
721 | 第三类前列腺素 PGE3 | 1514 | 顺-11-二十碳烯酸 cis-gondoic acid |
1495 | 前列腺素 PGE2 | 1366 | 顺式-11-二十碳烯酸 cis-9-palmitoleic acid |
1435 | 前列腺素 E1PGE1 | 454 | 胆酸 Cholic acid |
1661 | 棕榈酸 Palmitic acid | 1758 | 肾上腺酸 Adrenic Acid |
1375 | 十八烷二酸 Octadecanedioic acid | 769 | (5Z,7E,11Z,14Z)-9-羟基-5,7,11,14-二十碳四烯酸 9-HETE |
1705 | 肉豆蔻油酸 Myristoleic acid | 534 | (8R)-8-氢过氧基亚油酸 8R-HpODE |
1821 | 1-羟基-2-肉豆蔻酰-sn-甘油-3-磷酸乙醇胺 LysoPE(0:0/14:0) | 680 | L-4-羟基色氨酸 4-Hydroxy-L-threonine |
1109 | L-脯氨酰-L-苯丙氨酸 L-prolyl-L-phenylalanine | 1859 | 3-吲哚丙酸3-Indolepropionic acid |
467 | 1-(1Z十六碳烯酰基)-sn-甘油-3-磷酸 LPA(P-16:0e/0:0) | 1177 | 3-吲哚丁酸 3-Indolebutyric acid |
1106 | 2棕榈酰甘油-3-磷酸 LPA(0:0/16:0) |
表4
正离子模式下VIP得分表"
编号 No. | 胸肌Breast muscle | 编号 No. | 腿肌Thigh muscle | ||
---|---|---|---|---|---|
Var ID (Primary) | VIP pred | Var ID (Primary) | VIP pred | ||
1 | 肌酸 Creatine | 7.01967 | 1 | 肌酸 Creatine | 6.56079 |
2 | L-鹅肌肽 L-Anserine | 4.57094 | 2 | 肌酐 Creatinine | 3.23688 |
3 | 甜菜碱 Betaine | 2.84563 | 3 | 左旋肉碱 L-Carnitine | 3.22422 |
4 | 1-甲基组氨酸1-Methylhistidine | 1.83296 | 4 | 甜菜碱 Betaine | 3.20225 |
5 | N-[4-(乙酰基氨基)苯基]-3-氧代丁酰胺Hydrouracil | 1.64535 | 5 | L-鹅肌肽 L-Anserine | 3.00584 |
6 | 1-棕榈酰-2-羟基-sn-甘油-3-磷酸胆碱 LysoPC(16:0) | 1.48292 | 6 | 3-(硬脂酰氧基)-4-(三甲基铵基)丁酸酯DL-Stearoylcarnitine | 1.81764 |
7 | 肌苷 Inosine | 1.39204 | 7 | L-谷氨酰胺 L-Glutamine | 1.63342 |
8 | L-色氨酸 L-Tryptophan | 1.37077 | 8 | 次黄嘌呤 Hypoxanthine | 1.52174 |
9 | 5'-甲基硫腺苷5'-Methylthioadenosine | 1.23907 | 9 | N-乙酰-L-组氨酸N-Acetyl-L-Histidine | 1.44579 |
10 | N-乙酰-L-组氨酸N-Acetyl-L-Histidine | 1.22247 | 10 | L-组氨酸 L-Histidine | 1.31488 |
11 | 异亮脯氨酸 Isoleucylproline | 1.20120 | 11 | 乙酰肉碱Acetylcarnitine | 1.22599 |
12 | 肌酐 Creatinine | 1.11686 | 12 | 1-棕榈酰-2-羟基-sn-甘油-3-磷酸胆碱LysoPC(16:0) | 1.14697 |
表5
负离子模式下VIP得分表"
编号 No. | 胸肌Breast muscle | 编号 No. | 腿肌Thigh muscle | ||
---|---|---|---|---|---|
Var ID (Primary) | VIP pred | Var ID (Primary) | VIP pred | ||
1 | 苏糖酸 Threonic acid | 2.77134 | 1 | L-4-羟基色氨酸4-Hydroxy-L-threonine | 2.76987 |
2 | 景天庚酮糖 Sedoheptulose | 2.69728 | 2 | L-谷氨酰5-磷酸酯 L-Glutamyl 5-phosphate | 2.14000 |
3 | L-4-羟基色氨酸4-Hydroxy-L-threonine | 2.38874 | 3 | 景天庚酮糖 Sedoheptulose | 2.04383 |
4 | 尿苷 Uridine | 2.21336 | 4 | 苏糖酸 Threonic acid | 1.94052 |
5 | D-葡萄糖-6-磷酸二钠D-Glucose 6-phosphate | 1.51438 | 5 | 3-吲哚丁酸 3-Indolebutyric acid | 1.58979 |
6 | γ-谷氨酰谷氨酸 Gamma Glutamylglutamic acid | 1.31314 | 6 | D-葡萄糖 6-磷酸二钠D-Glucose 6-phosphate | 1.57744 |
7 | 1-(1Z十六碳烯酰基)-sn-甘油-3-磷酸LPA(P-16:0e/0:0) | 1.30657 | 7 | 前列腺素 PGE2 | 1.3055 |
8 | 吲哚乳酸 Indolelactic acid | 1.12537 | 8 | 脯氨酰羟脯氨酸Prolylhydroxyproline | 1.27561 |
9 | 脯氨酰羟脯氨酸Prolylhydroxyproline | 1.08437 | 9 | γ-谷氨酰谷氨酸 Gamma Glutamylglutamic acid | 1.19071 |
10 | 十一烷二酸 Undecanedioic acid | 1.01416 | 10 | 1-(1Z十六碳烯酰基)-sn-甘油-3-磷酸LPA(P-16:0e/0:0) | 1.01964 |
11 | 谷氨酰丙氨酸 Glutamylalanine | 1.00139 |
表6
茶花鸡肌肉中呈味物质和TAV值"
呈味 Taste | 名称 Name | 阈值[ Threshold (mmoL∙L-1) | 原料胸肉 Raw breast muscle | 熟胸肉 Cooked breast muscle | 原料腿肉 Raw thigh muscle | 熟腿肉 Cooked thigh muscle | ||||
---|---|---|---|---|---|---|---|---|---|---|
含量 (mmoL∙L-1) | TAV | 含量 (mmoL∙L-1) | TAV | 含量 (mmoL∙L-1) | TAV | 含量 (mmoL∙L-1) | TAV | |||
酸涩味Sour | 苹果酸Malic acid | 3.70 | 0.03±0.00a | 0.01 | 0.02±0.00b | 0.01 | 0.02±0.00b | 0.01 | 0.02±0.00b | 0.01 |
酸涩味Sour | γ-氨基丁酸 γ-Aminobutyric acid | 0.02 | 0.24±0.05c | 12.00 | 0.46±0.06b | 23.00 | 0.70±0.10a | 35.00 | 0.75±0.13a | 37.50 |
其他氨基酸OAA | 0.27±0.05c | 0.48±0.07b | 0.72±0.10a | 0.77±0.13a | ||||||
苦味Bitter | 色氨酸 L-Tryptophan | 5.00 | 1.50±0.30a | 0.30 | 1.18±0.19a | 0.24 | 1.17±0.15a | 0.23 | 0.32±0.03b | 0.06 |
苦味Bitter | 次黄嘌呤Hypoxanthine | 9.00 | 1.21±0.14b | 0.13 | 5.91±2.08a | 0.66 | 2.47±0.02b | 0.27 | 2.45±0.62b | 0.27 |
苦味Bitter | 异亮氨酸L-Isoleucine | 11.00 | 0.52±0.15b | 0.05 | 0.56±0.13b | 0.05 | 0.80±0.22a | 0.07 | 0.24±0.02c | 0.02 |
苦味Bitter | 酪氨酸L-Tyrosine | 5.00 | 0.89±0.05 | 0.18 | 0.69±0.10 | 0.14 | 0.71±0.10 | 0.14 | 0.94±0.27 | 0.19 |
苦味Bitter | 精氨酸L-Arginine | 2.87 | 0.38±0.11a | 0.13 | 0.21±0.03b | 0.07 | 0.43±0.06a | 0.15 | 0.39±0.11a | 0.14 |
苦味Bitter | 组氨酸L-Histidine | 3.22 | 3.03±0.02ab | 0.94 | 3.69±0.86a | 1.14 | 2.46±0.62bc | 0.76 | 1.82±0.54c | 0.56 |
苦味Bitter | 苯丙氨酸L-Phenylalanine | 3.03 | 0.32±0.07a | 0.11 | 0.43±0.08a | 0.14 | 0.36±0.10a | 0.12 | 0.11±0.06b | 0.04 |
苦味氨基酸BFA | 7.89±0.43b | 12.14±2.00a | 8.77±0.88b | 6.27±1.25b | ||||||
甜味Sweet | 脯氨酸L-Proline | 26.00 | 0.67±0.18b | 0.03 | 0.82±0.12b | 0.03 | 0.81±0.11b | 0.03 | 1.54±0.50a | 0.06 |
甜味Sweet | 丝氨酸L-Serine | 30.00 | 0.08±0.01b | 0.00 | 0.02±0.00c | 0.00 | 0.66±0.06a | 0.02 | 0.08±0.02b | 0.00 |
甜味Sweet | 苏氨酸L-Threonine | 40.00 | 0.12±0.03 | 0.00 | 0.11±0.03 | 0.00 | 0.11±0.01 | 0.00 | 0.12±0.05 | 0.00 |
甜味Sweet | D-葡萄糖6-磷酸 D-Glucose 6-phosphate | 48.00 | 9.21±2.29a | 0.19 | 9.42±1.96a | 0.20 | 1.65±0.19b | 0.03 | 1.59±0.52b | 0.03 |
甜味Sweet | 甜菜碱Betaine | 20.01 | 4.58±1.02c | 0.23 | 5.90±0.89c | 0.29 | 14.03±3.66b | 0.70 | 39.98±6.38a | 2.00 |
甜味氨基酸SFA | 14.74±2.1b | 16.36±1.37b | 17.34±3.81b | 43.30±7.22a | ||||||
鲜味Umami | 腺甘酸 Adenosine monophosphate | 4.00 | 0.01±0.00 | 0.00 | 0.01±0.00 | 0.00 | 0.01±0.00 | 0.00 | 0.01±0.00 | 0.00 |
鲜味Umami | 谷氨酸盐L-Glutamate | 3.40 | 0.03±0.01b | 0.01 | 0.07±0.01a | 0.02 | 0.08±0.02a | 0.02 | 0.04±0.02b | 0.01 |
鲜味Umami | 肌苷酸IMP | 0.68 | 0.11±0.02ab | 0.16 | 0.07±0.02b | 0.10 | 0.12±0.03a | 0.18 | 0.08±0.03ab | 0.12 |
鲜味氨基酸UFA | 0.14±0.02b | 0.15±0.02b | 0.22±0.05a | 0.13±0.05b |
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