[논문]Metabolomic profiling of postmortem aged muscle in Japanese Brown beef cattle revealed an interbreed difference from Japanese Black beef

Susumu Muroya; Riko Nomura; Hirotaka Nagai; Koichi Ojima; Kazutsugu Matsukawa

doi:10.5713/ab.22.0202

[국내논문] Metabolomic profiling of postmortem aged muscle in Japanese Brown beef cattle revealed an interbreed difference from Japanese Black beef 원문보기

Animal bioscience, v.36 no.3, 2023년, pp.506 - 520

Susumu Muroya (Animal Products Research Division, NARO Institute of Livestock and Grassland Science) , Riko Nomura (Kochi Prefectural Seibu Animal Hygiene Service Center) , Hirotaka Nagai (Department of Agriculture and Marine Science, Kochi University) , Koichi Ojima (Animal Products Research Division, NARO Institute of Livestock and Grassland Science) , Kazutsugu Matsukawa (Department of Agriculture and Marine Science, Kochi University)

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Objective: Japanese Brown (JBR) cattle, especially the Kochi (Tosa) pedigree (JBRT), is a local breed of moderately marbled beef. Despite the increasing demand, the interbreed differences in muscle metabolites from the highly marbled Japanese Black (JBL) beef remain poorly understood. We aimed to determine flavor-related metabolites and postmortem metabolisms characteristic to JBRT beef in comparison with JBL beef. Methods: Lean portions of the longissimus thoracis (loin) muscle from four JBRT cattle were collected at 0, 1, and 14 d postmortem. The muscle metabolomic profiles were analyzed using capillary electrophoresis time-of-flight mass spectrometry. The difference in post-mortem metabolisms and aged muscle metabolites were analyzed by statistical and bioinformatic analyses between JBRT (n = 12) and JBL cattle (n = 6). Results: A total of 240 metabolite annotations were obtained from the detected signals of the JBRT muscle samples. Principal component analysis separated the beef samples into three different aging point groups. According to metabolite set enrichment analysis, post-mortem metabolic changes were associated with the metabolism of pyrimidine, nicotinate and nicotinamide, purine, pyruvate, thiamine, amino sugar, and fatty acid; citric acid cycle; and pentose phosphate pathway as well as various amino acids and mitochondrial fatty acid metabolism. The aged JBRT beef showed higher ultimate pH and lower lactate content than aged JBL beef, suggesting the lower glycolytic activity in postmortem JBRT muscle. JBRT beef was distinguished from JBL beef by significantly different compounds, including choline, amino acids, uridine monophosphate, inosine 5'-monophosphate, fructose 1,6-diphosphate, and betaine, suggesting interbreed differences in the accumulation of nucleotide monophosphate, glutathione metabolism, and phospholipid metabolism. Conclusion: Glycolysis, purine metabolism, fatty acid catabolism, and protein degradation were the most common pathways in beef during postmortem aging. The differentially expressed metabolites and the relevant metabolisms in JBRT beef may contribute to the development of a characteristic flavor.

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표/그림 (13)

그림 Figure 2. Results of principal component analysis (PCA, panel A) and analysis of variance (ANOVA, panel B) of metabolomic profiles during postmortem aging in Japanese Brown longissimus thoracis (LT) muscle. In panel A, the circle, square, and triangle symbols indicate LT samples taken at 0 (D0), 24 h (D1), and 168 h (D14) postmortem, respectively. Red (F), heifer; blue (M), steer.
그림 Figure 1. Postmortem pH decline in the longissimus thoracis (LT) muscle of Japanese Brown (black column) and Japanese Black (gray column) cattle. * and ** indicate significant difference between the breeds at p<0.05 and <0.01, respectively. Error bars indicate standard error of the mean.
표 Table 1. Loadings for PC1 and PC2 in the principal component analysis
그림 Figure 3. Heatmap result of hierarchical cluster analysis (HCA) of metabolomic changes during postmortem aging in the longissimus thoracis (LT) muscle of Japanese Brown cattle. The four cattle samples indicated under the graphic were allocated to each group at a specific time point (day 0, D0; day 1, D1; day 14, D14). The row displays the metabolite, and the column represents the sample. Metabolites with relatively low and high levels are displayed in light blue and brown, respectively. The brightness of each color corresponds to the magnitude of the difference when compared to the average value. F, heifer; M, steer.
그림 Figure 4. Postmortem changes in metabolite levels in the longissimus thoracis (LT) muscle of Japanese Brown cattle. The levels (vertical axis) are indicated in nmol/g. The horizontal axis indicates postmortem time (day 0, day 1, day 14).
그림 Figure 5. Result of metabolite set enrichment analysis for postmortem metabolite changes (day 0 [D0] versus day 14 [D14]) in the longissimus thoracis (LT) muscle of Japanese Brown cattle. Enrichment ratio is computed by hits/expected, where hits = observed hits; expected = expected hits.
그림 Figure 6. Heatmap result of hierarchical cluster analysis (HCA) of metabolomic changes during postmortem aging in the longissimus thoracis (LT) muscle of Japanese Black (JBL) and Japanese Brown (JBRT) cattle. The three JBL and four JBRT samples at a specific time point (day 0, D0; day 1, D1; day 14, D14) indicated under the heatmap were allocated to each breed group. The row displays the metabolite, and the column represents the sample. Metabolites with relatively low and high levels are displayed in light blue and brown respectively. The brightness of each color corresponds to the magnitude of the difference when compared to the average value.
그림 Figure 7. Postmortem changes in metabolite levels in the longissimus thoracis (LT) muscle of Japanese Black and Japanese Brown cattle. The levels (vertical axis) are indicated in nmol/g. The horizontal axis indicates postmortem time (day 0, day 1, day 14). Amino acids (AAs) are indicated by their three letter representations.
그림 Figure 8. Differences in metabolite levels between Japanese black (JBL) and Japanese Brown (JBRT) longissimus thoracis (LT) muscles at day 14 postmortem. (A) Volcano plot. Horizontal axis indicates log2 fold change (JBRT/JBL ratio) for each metabolite, and vertical axis indicates -log10(P-value). Red, increased in JBRT; blue, decreased in JBRT. (B) Relative levels of metabolites in JBL and JBRT beef samples. Vertical axis indicates relative level normalized by autoscaling.
그림 Figure 9. Results of principal component analysis (PCA) for metabolome profiles of postmortem Japanese Black (JBL, red) and Japanese Brown (JBRT, green) beef samples at day 14 postmortem.
그림 Figure 10. Heatmap result of hierarchical cluster analysis (HCA) of Japanese Black (JBL) and Japanese Brown (JBRT) beef samples at day 14 postmortem. The six JBL (red) and 12 JBRT (green) samples indicated under the heatmap were allocated to each breed group. The row displays the metabolite, and the column represents the sample. Metabolites with relatively low and high levels are displayed in light blue and brown, respectively. The brightness of each color corresponds to the magnitude of the difference when compared to the average value.
그림 Figure 11. Result of partial least squares-discriminant analysis (PLS-DA) for metabolomic difference between Japanese Black and Japanese Brown beef at day 14 postmortem. (A) Score plot of component-1 and -2. (B) Variable importance in projection (VIP) scores of top 30 metabolites.
그림 Figure 12. Result of metabolite set enrichment analysis for metabolomic difference between Japanese Black and Japanese Brown beef at day 14 postmortem. Enrichment ratio is computed by hits/Expected, where hits = observed hits; expected = expected hits.

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