宰后不同时间滩羊肉抗氧化活性的变化及可能机制

doi:10.3864/j.issn.0578-1752.2021.23.015

Abstract

Abstract:

【Objective】The aim of this study was to explore the changes of antioxidant activity of Tan sheep meat at different time points after slaughter, and to explain its mechanism from the perspective of free amino acids and proteome, so as to provide the data support for the quality maintenance of fresh mutton. 【Method】Six 6-month-old healthy stall-feeding male lambs (18.30±1.41 kg) were selected in this study. The longissimus dorsi of the lambs was collected at 0.5, 3, 6, 12 and 48 h after slaughter to determine the ferric ion reducing antioxidant power (FRAP), free radical scavenging capacity of 2,2-azinobis-(3-ethylbenzothiazoline-6-sulphonate) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH), oxygen-radical absorbance capacity (ORAC) of DPPH, total antioxidant capacity of N,N-dimethyl-p-phenylendiamine (DMPD), content of free amino acids and the changes of proteomic. 【Result】The results showed that within 48 h postmortem, the FRAP total antioxidant capacity decreased first and then increased. While the free radical scavenging capacity of ABTS increased first and then tended to be stable. The free radical scavenging capacity and ORAC of DPPH and DMPD all showed an upward trend. Total antioxidant capacity of FRAP, free radical scavenging capacity and ORAC of ABTS, DPPH, DMPD of Tan sheep meat at 48 h after slaughter were all significantly higher than those at 0.5 and 3 h after slaughter (P<0.05). No significant difference was found in total content of free amino acids (P>0.05), but the contents of Cys, Leu, Tyr, Ile, Phe and Arg in Tan sheep meat were significantly different among different time points after slaughter (P<0.05), and showed an increase trend. There were 7 free amino acids positively correlated with FRAR, including o-phosphoserine (r = 0.489, P<0.01), leucine (r=0.566, P<0.01), tyrosine (r=0.596, P<0.01), isoleucine (r=0.374, P<0.05), phenylalanine (r=0.499, P<0.01), lysine (r=0.375, P<0.05) and arginine (r=0.376, P<0.05). Among them, Leu, Tyr, Phe and Arg were significantly positively correlated with FRAP, ABTS, DPPH, ORAC and DMPD (P<0.05). Twenty differential proteins in Tan sheep meat at different time points after slaughter were screened and identified by proteomic. There were 12 differential proteins significantly correlated with FRAP (P<0.05). Calpain inhibitor protein and unknown protein 2 were significantly negatively correlated with FRAP, ABTS, DPPH, ORAC and DMPD (P<0.05). Glycogen protein 1 and protein kinase domain proteins were significantly positively correlated with FRAP, ABTS, DPPH, ORAC and DMPD (P<0.05). 【Conclusion】This research indicated that the total antioxidant capacity of Tan sheep meat increased within 48 h after slaughter, which was related to the release of free amino acids such as Leu, Tyr, Phe and Arg in the postmortem muscle. The decrease protein expression of calpain inhibitor protein and unknown protein 2, and the increased protein expression of glycogen protein 1 and protein kinase domain protein in muscle were significantly correlated with the changes of antioxidant capacity of Tan sheep meat after slaughter.

Key words: postmortem, antioxidant activity, free amino acids, proteome, lamb

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.

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Figures/Tables 8

Fig. 1

Fig. 2

Table 1

Free amino acid contents of Tan lamb in different postmortem time"

氨基酸含量 Amino acid (mg/100 g fresh meat)	宰后时间Postmortem time (h)					P值 P value
氨基酸含量 Amino acid (mg/100 g fresh meat)	0.5	3	6	12	48	P值 P value
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

Table 1

Table 2

Fig. 3

Fig. 4

Fig. 5

Table 3

Person’s correlation between differentially abundant proteins and antioxidant activity in Tan lamb"

登记号 Accession	蛋白名称 Protein name	相关系数 Correlation coefficient
登记号 Accession	蛋白名称 Protein name	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**

Table 3

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氨基酸名称 Amino acid name	相关系数 Correlation coefficients
氨基酸名称 Amino acid name	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

Changes of Antioxidant Activity and Its Possible Mechanism in Tan Sheep Meat in Different Postmortem Time

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