中国农业科学 ›› 2021, Vol. 54 ›› Issue (23): 5110-5124.doi: 10.3864/j.issn.0578-1752.2021.23.015
侯成立1(),黄彩燕1,郑晓春1,刘维华2,杨奇2(),张德权1()
收稿日期:
2021-05-17
接受日期:
2021-08-10
出版日期:
2021-12-01
发布日期:
2021-12-06
通讯作者:
杨奇,张德权
作者简介:
侯成立,E-mail: 基金资助:
HOU ChengLi1(),HUANG CaiYan1,ZHENG XiaoChun1,LIU WeiHua2,YANG Qi2(),ZHANG DeQuan1()
Received:
2021-05-17
Accepted:
2021-08-10
Online:
2021-12-01
Published:
2021-12-06
Contact:
Qi YANG,DeQuan ZHANG
摘要:
【目的】探究宰后不同时间滩羊肉抗氧化活性的变化,并从游离氨基酸、蛋白质组角度阐释其机制,为生鲜羊肉的品质保持提供数据支持。【方法】选取6只6月龄健康舍饲滩羊公羊((18.30±1.41)kg),在宰后0.5、3、6、12和48 h取背最长肌,测定铁离子还原/抗氧化能力(ferric ion reducing antioxidant power,FRAP)、2, 2-联氮基双-(3-乙基苯并噻唑啉-6-磺酸)二铵盐(2, 2-azinobis-(3- ethylbenzothiazoline-6-sulphonate),ABTS)自由基清除能力、2, 2-二苯代苦味酰基苯肼(2, 2-diphenyl-1-picrylhydrazyl,DPPH)自由基清除能力、氧自由基吸收能力(oxygen-radical absorbance capacity,ORAC)、N, N-二甲基-对苯二胺(N, N-dimethyl-p-phenylendiamine,DMPD)总抗氧化能力、游离氨基酸含量和蛋白质组变化。【结果】宰后48 h内,滩羊肉中FRAP总抗氧化能力呈先下降后上升趋势,ABTS自由基清除能力呈先升高后趋于稳定趋势,DPPH、DMPD自由基清除能力和ORAC整体均呈现上升趋势;宰后48 h滩羊肉中FRAP总抗氧化能力、ABTS、DPPH、DMPD自由基清除能力和ORAC显著高于宰后0.5 h和3 h水平(P<0.05);宰后不同时间点滩羊肉中游离氨基酸总含量差异不显著(P>0.05),但胱氨酸、亮氨酸、酪氨酸、异亮氨酸、苯丙氨酸、精氨酸的含量在不同时间点存在显著性差异(P<0.05),且总体呈现增加的趋势。与FRAP呈显著正相关关系的游离氨基酸有7个,包括邻磷酸丝氨酸(r=0.489,P<0.01)、亮氨酸(r=0.566,P<0.01)、酪氨酸(r=0.596,P<0.01)、异亮氨酸(r=0.374,P<0.05)、苯丙氨酸(r=0.499,P<0.01)、赖氨酸(r=0.375,P<0.05)和精氨酸(r=0.376,P<0.05);其中,亮氨酸、酪氨酸、苯丙氨酸、精氨酸与FRAP、ABTS、DPPH、ORAC、DMPD均呈现显著正相关关系(P<0.05)。蛋白组学鉴定筛选出宰后不同时间滩羊肉中差异蛋白20个,与FRAP显著相关的差异蛋白有12个(P<0.05),钙蛋白酶抑制蛋白和未知蛋白2与FRAP、ABTS、DPPH、ORAC、DMPD均呈现显著负相关关系(P<0.05),而糖原蛋白1和蛋白激酶结构域蛋白与FRAP、ABTS、DPPH、ORAC、DMPD均呈现显著正相关关系(P<0.05)。【结论】滩羊肉抗氧化能力的升高与宰后肌肉中亮氨酸、酪氨酸、苯丙氨酸、精氨酸等游离氨基酸的释放有关。宰后初期肌肉中钙蛋白酶抑制蛋白、未知蛋白2等蛋白表达量的下降和糖原蛋白1、蛋白激酶结构域蛋白等蛋白表达量的上升与宰后滩羊肉抗氧化能力的变化显著相关。
侯成立,黄彩燕,郑晓春,刘维华,杨奇,张德权. 宰后不同时间滩羊肉抗氧化活性的变化及可能机制[J]. 中国农业科学, 2021, 54(23): 5110-5124.
HOU ChengLi,HUANG CaiYan,ZHENG XiaoChun,LIU WeiHua,YANG Qi,ZHANG DeQuan. Changes of Antioxidant Activity and Its Possible Mechanism in Tan Sheep Meat in Different Postmortem Time[J]. Scientia Agricultura Sinica, 2021, 54(23): 5110-5124.
表1
宰后不同时间滩羊肉中游离氨基酸含量"
氨基酸含量 Amino acid (mg/100 g fresh meat) | 宰后时间Postmortem time (h) | P值 P value | ||||
---|---|---|---|---|---|---|
0.5 | 3 | 6 | 12 | 48 | ||
邻磷酸丝氨酸 o-Phospho-serine | 4.16±0.29b | 4.14±0.31b | 4.19±0.64b | 4.45±0.41ab | 4.85±0.65a | 0.08 |
牛磺酸 Taurine | 79.10±41.24a | 92.20±32.03a | 80.19±28.93a | 73.62±27.64a | 70.26±36.18a | 0.83 |
邻苯二乙醇胺 o-Phsophoethanolamine | 3.25±0.68a | 3.18±0.34a | 2.95±0.39a | 2.80±0.42a | 3.09±0.71a | 0.60 |
天冬氨酸 Aspartic acid | 0.76±1.19a | 0.50±1.23a | 0.05±0.13a | 0.00±0.00a | 0.04±0.09a | 0.33 |
苏氨酸 Threonine | 4.33±1.53a | 5.03±1.19a | 4.74±1.69a | 5.05±1.33a | 5.22±1.24a | 0.82 |
丝氨酸 Serine | 6.63±0.47a | 6.68±0.71a | 6.76±0.63a | 6.92±1.18a | 6.78±0.72a | 0.97 |
谷氨酸 Glutamic acid | 120.10±15.06a | 111.63±19.4a | 108.09±22.41a | 111.07±25.38a | 108.38±24.75a | 0.87 |
甘氨酸 Glycine | 20.13±3.15a | 21.22±2.50a | 20.69±2.42a | 21.40±1.18a | 20.88±1.92a | 0.90 |
丙氨酸 Alanine | 49.53±6.12a | 51.46±4.48a | 51.76±4.34a | 52.07±2.27a | 51.04±2.23a | 0.85 |
缬氨酸 Valine | 1.39±0.85a | 0.92±0.44a | 0.97±1.31a | 1.30±0.82a | 1.37±0.28a | 0.77 |
蛋氨酸 Methionine | 0.04±0.02a | 0.03±0.01a | 0.04±0.02a | 0.05±0.03a | 0.05±0.05a | 0.84 |
胱氨酸 Cystine | 1.04±0.10b | 1.17±0.10ab | 1.14±0.14b | 1.09±0.17b | 1.31±0.16a | 0.02 |
亮氨酸 Leucine | 1.81±0.30c | 2.13±0.17bc | 2.20±0.13b | 2.27±0.19b | 2.66±0.46a | 0.00 |
酪氨酸 Tyrosine | 4.97±0.67c | 5.78±0.75bc | 6.17±0.66b | 6.42±0.72b | 7.55±1.05a | 0.00 |
异亮氨酸 Isoleucine | 1.99±0.29b | 2.40±0.43ab | 2.43±0.37ab | 2.47±0.31ab | 2.79±0.53a | 0.03 |
苯丙氨酸 Phenylalanine | 2.00±0.40c | 2.33±0.31bc | 2.41±0.27bc | 2.49±0.32ab | 2.89±0.43a | 0.00 |
γ-氨基丁酸 γ-Amino-n-butyric acid | 0.90±0.16a | 0.85±0.17a | 0.88±0.14a | 0.86±0.10a | 0.80±0.07a | 0.78 |
氨 Ammonia | 18.83±4.17a | 20.57±2.66a | 20.99±2.15a | 21.08±1.71a | 21.75±1.94a | 0.42 |
鸟氨酸 Ornithine | 1.70±0.53a | 1.38±0.34bc | 1.27±0.34bc | 1.10±0.12b | 1.49±0.42bc | 0.10 |
赖氨酸 Lysine | 2.56±0.91b | 3.12±0.58ab | 3.21±0.66ab | 3.31±0.57ab | 3.79±0.89a | 0.10 |
组氨酸 Histidine | 2.57±0.36a | 2.80±0.73a | 2.84±0.92a | 3.14±0.69a | 3.32±0.70a | 0.40 |
鹅肌肽 Anserine | 197.54±25.69a | 200.95±24.56a | 208.89±18.77a | 213.93±18.28a | 202.07±28.81a | 0.76 |
肌肽 Carnosine | 499.15±27.17a | 530.58±65.12a | 533.56±85.88a | 555.97±34.04a | 528.04±65.02a | 0.60 |
精氨酸 Arginine | 6.03±1.02c | 7.03±0.81bc | 7.53±1.41ab | 7.13±1.05ab | 8.51±0.63a | 0.01 |
脯氨酸 Proline | 5.63±0.89ab | 4.88±1.62b | 5.29±1.07ab | 5.20±0.62ab | 6.32±0.89a | 0.15 |
合计 Total | 1086.05±62.03a | 1100.05±78.03a | 1081.83±70.54a | 1100.43±15.42a | 1085.16±57.00a | 0.97 |
表2
滩羊肉中游离氨基酸含量与抗氧化活性的相关性"
氨基酸名称 Amino acid name | 相关系数 Correlation coefficients | ||||
---|---|---|---|---|---|
FRAP | ABTS | DPPH | ORAC | DMPD | |
邻磷酸丝氨酸 o-Phospho-serine | 0.480** | 0.208 | 0.420* | 0.417* | 0.417* |
牛磺酸 Taurine | -0.226 | -0.091 | -0.188 | -0.227 | -0.235 |
邻苯二乙醇胺 o-Phsophoethanolamine | -0.106 | -0.236 | -0.222 | -0.132 | -0.181 |
天冬氨酸 Aspartic acid | -0.252 | -0.393* | -0.378* | -0.246 | -0.268 |
苏氨酸 Threonine | 0.128 | 0.213 | 0.135 | 0.192 | 0.241 |
丝氨酸 Serine | 0.114 | 0.175 | 0.120 | 0.143 | 0.137 |
谷氨酸 Glutamic acid | -0.034 | -0.129 | -0.162 | -0.117 | -0.110 |
甘氨酸 Glycine | 0.063 | 0.083 | 0.062 | -0.040 | 0.086 |
丙氨酸 Alanine | 0.077 | 0.182 | 0.113 | 0.076 | 0.098 |
缬氨酸 Valine | 0.168 | -0.109 | 0.111 | 0.208 | 0.082 |
蛋氨酸 Methionine | 0.270 | -0.022 | 0.168 | 0.247 | 0.080 |
胱氨酸 Cystine | 0.313 | 0.227 | 0.296 | 0.226 | 0.312 |
亮氨酸 Leucine | 0.566** | 0.487** | 0.597** | 0.471** | 0.629** |
酪氨酸 Tyrosine | 0.596** | 0.493** | 0.656** | 0.532** | 0.679** |
异亮氨酸 Isoleucine | 0.374* | 0.391* | 0.456* | 0.324 | 0.425* |
苯丙氨酸 Phenylalanine | 0.499** | 0.405* | 0.545** | 0.445* | 0.569** |
γ-氨基丁酸 γ-Amino-n-butyric acid | -0.234 | -0.171 | -0.144 | -0.193 | -0.153 |
氨 Ammonia | 0.235 | 0.371* | 0.289 | 0.230 | 0.258 |
鸟氨酸 Ornithine | -0.010 | -0.402* | -0.274 | -0.117 | -0.091 |
赖氨酸 Lysine | 0.375* | 0.265 | 0.410* | 0.271 | 0.391* |
组氨酸 Histidine | 0.333 | 0.252 | 0.337 | 0.300 | 0.372* |
鹅肌肽 Anserine | 0.075 | 0.269 | 0.157 | 0.073 | 0.095 |
肌肽 Carnosine | 0.040 | 0.370* | 0.187 | 0.110 | 0.170 |
精氨酸 Arginine | 0.376* | 0.486** | 0.493** | 0.384* | 0.536** |
脯氨酸 Proline | 0.324 | -0.078 | 0.212 | 0.285 | 0.370* |
合计 Total | -0.089 | -0.014 | -0.016 | -0.116 | -0.078 |
表3
滩羊肉中差异蛋白丰度与抗氧化活性的相关性"
登记号 Accession | 蛋白名称 Protein name | 相关系数 Correlation coefficient | ||||
---|---|---|---|---|---|---|
FRAP | ABTS | DPPH | ORAC | DMPD | ||
C3V6M4 | 钙蛋白酶抑制蛋白 Calpastatin | -0.556* | -0.587* | -0.638** | -0.573* | -0.589* |
W5NRQ5 | 载脂蛋白B mRNA编辑酶催化亚基2 Apolipoprotein B mRNA editing enzyme catalytic subunit 2 | -0.432 | -0.544* | -0.584* | -0.409 | -0.599** |
W5NS65 | H15结构域蛋白 H15 domain-containing protein | 0.471* | -0.155 | 0.142 | 0.396 | 0.375 |
W5NTS5 | 辅酶Q8A Coenzyme Q8A | -0.223 | -0.372 | -0.448 | -0.289 | -0.359 |
W5NUA0 | 脯氨酸/精氨酸丰富端亮氨酸丰富重复蛋白 Proline and arginine rich end leucine rich repeat protein | 0.052 | 0.541* | 0.422 | 0.126 | 0.169 |
W5P5C5 | 糖原蛋白1 Glycogenin 1 | 0.642** | 0.655** | 0.800** | 0.637** | 0.818** |
W5P8Z2 | 亲联蛋白1 Junctophilin 1 | -0.786** | -0.467 | -0.632** | -0.817** | -0.630** |
W5P9K6 | 蛋白酶体26S亚基ATP合酶 Proteasome 26S subunit, ATPase 3 | -0.738** | -0.384 | -0.558* | -0.722** | -0.651** |
W5PA76 | 亮氨酸拉链和CTNNBIP1结构域 Leucine zipper and CTNNBIP1 domain containing | -0.465* | -0.296 | -0.527 | -0.487* | -0.572* |
W5PCH1 | 胞膜窖相关蛋白4 Caveolae associated protein 4 | -0.442 | 0.168 | -0.100 | -0.333 | -0.305 |
W5PFV9 | 伴肌动蛋白 Nebulin | 0.627** | 0.253 | 0.539* | 0.587* | 0.596** |
W5PHT7 | 肽基脯氨酰异构酶 Peptidylprolyl isomerase | 0.562* | -0.027 | 0.256 | 0.508* | 0.416 |
W5PID4 | 未知蛋白1 Uncharacterized protein 1 | 0.025 | -0.502* | -0.319 | -0.056 | -0.151 |
W5PP64 | 未知蛋白2 Uncharacterized protein 2 | -0.557* | -0.674** | -0.674** | -0.627** | -0.673** |
W5PQJ1 | 真核细胞翻译起始因子4H Eukaryotic translation initiation factor 4H | -0.521* | -0.056 | -0.376 | -0.505* | -0.443 |
W5Q466 | 电压门控钙离子通道辅助亚基α2δ-1 Calcium voltage-gated channel auxiliary subunit alpha 2 delta 1 | 0.183 | 0.742** | 0.555* | 0.284 | 0.334 |
W5Q922 | 泛素羧基末端水解酶 Ubiquitin carboxyl-terminal hydrolase | 0.447 | -0.040 | 0.230 | 0.403 | 0.465 |
W5QG38 | 蛋白激酶结构域蛋白 Protein kinase domain-containing protein | 0.611** | 0.489* | 0.584* | 0.607** | 0.539* |
W5QGI4 | 苯丙氨酰-tRNA合成酶β Phenylalanyl-tRNA synthetase subunit beta | 0.261 | 0.634** | 0.499* | 0.456 | 0.406 |
W5QGM7 | H1组蛋白家族成员0 H1 histone family member 0 | 0.716** | 0.058 | 0.454 | 0.574* | 0.626** |
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