Untargeted metabolomics analysis of metabolic differences of crayfish (Procambarus clarkii) meat with different diets
-
摘要: 采用液相色谱-质谱联用非靶向代谢组学方法，研究了两种日粮饲喂下克氏原螯虾 (Procambarus clarkii) 肌肉代谢物的差异及变化，为提升其养殖品质提供参考。通过样本主成分分析 (Principal components analysis, PCA)，并与基因组百科全书数据库 (Kyoto Encyclopedia of Genes and Genomes, KEGG) 进行比对，筛选出虾肉中的差异代谢物并分析原因。结果显示：通过代谢组学分析，在正离子和负离子模式下共筛选出27种显著差异代谢物；与常规饲料相比，发酵饲料实验组中腺苷酸基琥珀酸、丝氨酸、磷酯酰胆碱、奎尼酸、补骨脂素、磷酯酰丝氨酸、谷氨酸等显著增加；经KEGG通路分析，变化显著的前４条通路分别是组氨酸代谢通路、精氨酸-脯氨酸代谢通路、蛋白质消化与吸收代谢通路和氨酰-tRNA合成通路。研究结果初步表明，日粮在调节克氏原螯虾氨基酸代谢、蛋白质合成和辅助合成氨酰-tRNA酶类等方面起积极作用。
-
关键词:
- 克氏原螯虾 /
- 非靶向代谢组学 /
- 液相色谱-质谱联用技术 /
- 差异代谢通路
Abstract: To provide references for improving the quality of crayfish (Procambarus clarkii) meat, we studied the differences in muscle metabolites of crayfish fed with two diets by a non-targeted metabolomic method using liquid chromatography mass spectrometry. Different metabolites in meat were screened by principal components analysis (PCA), cluster analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The results show that 27 significantly different metabolites were screened under the positive and negative ion modes. Compared with the control group (General feed), the contents of adenylsuccinic acid, phosphatidylcholine, quinate, psoralen, phosphatidylserine and glutamic in the experimental group (Fermented feed) increased significantly. According to KEGG pathway analysis, the top four pathways with the highest concentration of metabolites were histidine metabolism pathway, arginine proline metabolism pathway, protein digestion and absorption metabolism pathway, as well as -tRNA synthesis pathway. The results indicate that diets play a positive role in regulating amino acid metabolism, protein synthesis and assisting in the synthesis of aminoacyl-tRNA enzymes in organisms.-
Key words:
- Procambarus clarkii /
- Untargeted metabolomics /
- LC-MS /
- Differential metabolic pathways
-
图 1 发酵饲料组和常规饲料组代谢物的 PCA 分析图
Figure 1. PCA plots of metabolites in fermented feed group and general feed group
图 2 发酵饲料组和常规饲料组代谢物的 OPLS-DA 分析图
Figure 2. OPLS-DA plots of metabolites in fermented diet group and general diet group
图 3 差异代谢物的OPLS-DA置换检验结果
Figure 3. OPLS-DA model replacement test results of differential metabolites
图 4 发酵饲料组对比常规饲料组的差异代谢物筛选
Figure 4. Differential metabolite screening volcano maps of fermented diet group (T) and general diet group (CK)
图 5 发酵饲料组和常规饲料组 的层次聚类分析热力图
Figure 5. Hierarchical cluster analysis heat maps of fermented diet group (T) and general diet group (CK)
图 6 发酵饲料组对常规饲料组的通路分析气泡图
Figure 6. Path analysis bubble chart of fermented diet group and general diet group
表 1 基础饲料组成及营养水平 (干物质基础)
Table 1. Composition and nutrient levels of basal diet (Dry matter basis) %
项目
Item发酵饲料
Fermented diet常规饲料
General diet原料 Ingredient 鱼粉 Fish meal 6 16 豆粕 Soybean meal 50 12 菜籽粕 Rapeseed meal 12 15 亚麻粕 Flaxseed meal 8 0 玉米淀粉 Corn starch 8 28 小麦粉 Wheat flour 8 19 鱼油 Fish oil 5 3 豆油 Soybean oil 0 4 磷酸二氢钙 Ca(H2PO4)2 1.4 1.4 维生素① Vitamin 0.5 0.5 矿物质① Mineral 1 1 防霉剂 Mould inhibitor 0.1 0.1 合计 Total 100 100 营养水平 Nutrient Level 粗蛋白 Crude protein 32.00 32.00 粗脂肪 Crude fat 6 6 灰分 Ash 12 18 粗纤维 Crude fiber 14 8 赖氨酸 Lys 1.5 1.5 水分 Moisture 18 10 注：① 维生素和矿物质均为预混料，维生素主要含 VC、VE、VB、VD等，其他成分及含量保密。 Note: ① Vitamins and minerals are premixes, and vitamins mainly contain VC, VE, VB, VD, etc.. Other components and contents are confidential. 
下载: 导出CSV
表 2 不同饲料对克氏原螯虾生长性能的影响
Table 2. Effect of different feeds on growth performance of crayfish
指标
Index发酵饲料组
Fermented diet (T)常规饲料组
General diet (CK)初始体质量 IBM/g 5.95±0.42a 6.15±0.51a 终末体质量 FBM/g 28.29±2.86a 28.44±2.69a 体质量增长率 WGR/% 375.46±9.21a 362.40±8.88a 特定生长率 SGR/(%·d−1) 2.59±0.12a 2.55±0.19a 注：同行数据不同小写字母表示差异显著 (P<0.05)。 Note: Values with different letters within the same row are significantly different (P<0.05).
下载: 导出CSV
表 3 发酵饲料组和常规饲料组的显著差异代谢物列表
Table 3. List of significant different metabolites of fermented diet group and general diet group
模式
Mode差异代谢物
Differential metabolite变量投影
重要度
VIP差异倍数
FC质荷比
m/z显著性分析
P value保留时间
RT/s正离子 Positive ion 肌酸苷 Creatinine 3.41 0.32 114.06 0.0007 161.13 正离子 Positive ion 肌酸 Creatine 15.31 0.24 132.08 0.006 334.67 正离子 Positive ion 甲基组胺 Methyl-histamine 3.58 2.83 126.10 0.011 380.87 正离子 Positive ion 苦马豆素 Swainsonine 1.17 0.20 156.10 0.012 352.39 正离子 Positive ion 组氨酸 L-Histidine 7.81 0.39 156.08 0.015 441.29 正离子 Positive ion 精氨酸-谷氨酸 Arg-Glu 2.08 2.16 304.16 0.016 474.58 正离子 Positive ion 4-氨基吡啶 4-aminopyridine 2.78 2.94 95.06 0.024 377.41 正离子 Positive ion 腺苷酸琥珀酸 Adenylosuccinic acid 2.00 3.21 464.08 0.024 483.46 正离子 Positive ion 精氨酸 Arginine 24.77 0.74 175.12 0.027 507.05 正离子 Positive ion 磷脂酰胆碱 Lpc 16:0 6.96 2.01 496.34 0.029 183.71 正离子 Positive ion 戊二醛 Glutaraldehyde 1.82 0.58 83.06 0.029 350.75 正离子 Positive ion 鞘氨醇磷酰胆碱
N-oleoyl-d-erythro-sphingosylphosphorylcholine3.68 1.82 729.59 0.030 168.18 正离子 Positive ion 1-乙基-1,3-二氢-2H-苯并咪唑-2-酮
2h-benzimidazol-2-one,1-ethyl-1,3-dihydro-1.52 3.13 163.09 0.048 61.25 正离子 Positive ion 组氨-丝氨酸 His-Ser 1.83 2.12 243.11 0.044 330.91 正离子 Positive ion 肌氨酸 Sarcosine 1.89 2.49 134.01 0.044 338.91 正离子 Positive ion 3'-岩藻糖基乳糖 3'-fucosyllactose 1.62 17.32 114.06 0.048 391.17 正离子 Positive ion 辅酶 I Coenzyme I 2.22 0.61 664.11 0.049 423.70 负离子 Negative ion 3,4-二氯酚 3,4-dichlorophenol 1.11 1.43 160.94 0.014 315.87 负离子 Negative ion 奎尼酸 Quinate 1.39 14.33 191.05 0.019 330.78 负离子 Negative ion 丁基磷酸 Butylphosphonic acid 1.36 2.38 275.09 0.023 448.38 负离子 Negative ion 苹果酸 Malate 6.21 1.59 133.01 0.028 394.31 负离子 Negative ion 谷氨酸 L-Glutamate 1.01 1.80 168.03 0.029 385.65 负离子 Negative ion N-乙酰蛋氨酸 N-acetyl-l-methionine 1.45 2.49 190.05 0.029 187.41 负离子 Negative ion 磷酸丝氨酸 Phosphoserine 1.71 2.15 369.02 0.042 436.39 负离子 Negative ion 补骨脂定 Psoralidin 1.03 2.15 335.08 0.043 384.21 负离子Negative ion 东革内酯 Eurycomalactone 3.80 5.22 347.17 0.047 26.15 负离子 Negative ion 亚麻木酚素 Secoisolariciresinol 1.95 5.63 361.18 0.049 25.53
下载: 导出CSV
-
[1] 于秀娟, 郝向举, 党子乔, 等. 中国小龙虾产业发展报告 (2022)[J]. 中国水产, 2022(6): 47-54. doi: 10.3969/j.issn.1002-6681.2022.6.zhongguosc202206017 [2] 陈实, 吴旭干, 杨丰, 等. 配合饲料和传统饵料养殖脊尾白虾气味品质的比较[J]. 食品工业科技, 2020, 41(1): 189-194, 200. [3] CHANDRAN J, BELLAD A, RAMARAJAN M G, et al. Applications of quantitative metabolomics to revolutionize early diagnosis of inborn errors of metabolism in India[J]. Anal Sci Adv, 2021, 2(11/12): 546-563. [4] DUBUIS S, BAENKE F, SCHERBICHLER N, et al. Metabotypes of breast cancer cell lines revealed by non-targeted metabolomics[J]. Metab Eng, 2017, 43(Pt B): 173-186. [5] PAN Y, GU H W, LV Y, et al. Untargeted metabolomic analysis of Chinese red wines for geographical origin traceability by UPLC-QTOF-MS coupled with chemometrics[J]. Food Chem, 2022, 394: 133473. doi: 10.1016/j.foodchem.2022.133473 [6] WU CH C, MA Y J, WANG D, et al. Integrated microbiology and metabolomics analysis reveal plastic mulch film residue affects soil microorganisms and their metabolic functions[J]. J Hazard Mater, 2022, 423(PB): 127258. [7] 李玮, 贾婧怡, 李龙, 等. 核磁共振代谢组学技术鉴别天然奶油与人造奶油[J]. 食品科学, 2017, 38(12): 278-285. [8] 高淑芳, 张金鹏, 施永海, 等. 基于LC-MS技术的海、淡水养殖刀鲚卵巢的代谢组学比较分析[J]. 南方水产科学, 2022, 18(3): 68-75. doi: 10.12131/20210185 [9] 张舒, 王长远, 冯玉超, 等. 气相色谱-质谱联用代谢组学技术分析不同产地稻米代谢物[J]. 食品科学, 2021, 42(8): 206-213. [10] 林艳萍, 司端运, 刘昌孝. 液相色谱和质谱联用技术结合化学计量学应用于代谢组学的研究进展[J]. 分析化学, 2007(10): 1535-1540. [11] 刘慧茹, 汪海洋, 王喆, 等. 基于代谢组学和斑马鱼模型探究西洋参抗疲劳的关键活性成分[J]. 药学学报, 2023, 58(4): 1024-1032. [12] 张彦坤, 杨兵坤, 李航宇, 等. 饥饿胁迫下剑尾鱼肝脏代谢组学研究[J]. 四川动物, 2021, 40(6): 611-621. doi: 10.11984/j.issn.1000-7083.20210058 [13] 童铃, 金毅, 徐坤华, 等. 3种鲣鱼背部肌肉的营养成分分析及评价[J]. 南方水产科学, 2014, 10(5): 51-59. doi: 10.3969/j.issn.2095-0780.2014.05.008 [14] 虞为, 杨育凯, 林黑着, 等. 牛磺酸对花鲈生长性能、消化酶活性、抗氧化能力及免疫指标的影响[J]. 南方水产科学, 2021, 17(2): 78-86. [15] 叶彬清, 陶宁萍, 王锡昌. 秋刀鱼肌肉营养成分分析及评价[J]. 营养学报, 2014, 36(4): 406-408. [16] 邵俊杰, 钟立强, 朱昱璇, 等. 配合饲料和冰鲜鱼对大口黑鲈生长和品质的影响[J]. 水产科学, 2023, 42(1): 81-88. doi: 10.16378/j.cnki.1003-1111.21065 [17] WEN D L, LIU Y, YU Q. Metabolomic approach to measuring quality of chilled chicken meat during storage[J]. Poultry Sci, 2020, 99(5): 2543-2554. doi: 10.1016/j.psj.2019.11.070 [18] XU X Y, YANG H, XU Z, et al. The comparison of largemouth bass (Micropterus salmoides) fed trash fish and formula feeds: growth, flesh quality and metabolomics[J]. Front Nutr, 2022, 9: 966248. doi: 10.3389/fnut.2022.966248 [19] ZHANG K K, AI Q H, MAI K S, et al. Effects of dietary hydroxyproline on growth performance, body composition, hydroxyproline and collagen concentrations in tissues in relation to prolyl 4-hydroxylaseα (I) gene expression of juvenile turbot (Scophthalmus maximus L. ) fed high plant protein diets[J]. Aquaculture, 2013, 404/405: 77-84. doi: 10.1016/j.aquaculture.2013.04.025 [20] CARRAGHER J F, MÜHLHAUSLER B S, GEIER MS, et al. Effect of dietary ALA on growth rate, feed conversion ratio, mortality rate and breast meat omega-3 LCPUFA content in broiler chickens[J]. Anim Prod Sci, 2016, 56(5): 815-823. doi: 10.1071/AN14743 [21] MURUGESAN A, HOLMSTEDT S, BROWN K C, et al. Design and synthesis of novel quinic acid derivatives: in vitro cytotoxicity and anticancer effect on glioblastoma[J]. Future Med Chem, 2020(21): 1891-1910. [22] 李静, 陈冬梅, 陈明星. 亚麻木酚素小鼠体内抗氧化活性研究[J]. 中国粮油学报, 2012, 27(1): 48-52. doi: 10.3969/j.issn.1003-0174.2012.01.011 [23] BENEDETTA C, LUISA D M, CARLA C, et al. Sphingolipids and the immune system[J]. Pharmacol Res, 2003, 47(5): 421-437. doi: 10.1016/S1043-6618(03)00051-3 [24] SHIU W L, HUANG K R, HUNG J C, et al. Knockdown of zebrafish YY1a can downregulate the phosphatidylserine (PS) receptor expression, leading to induce the abnormal brain and heart development[J]. J Biomed Sci, 2016, 23(1): 31-33. doi: 10.1186/s12929-016-0248-1 [25] RUBÉN D, MIRIAN P, MOHAMMED G, et al. A comprehensive review on lipid oxidation in meat and meat products[J]. Antioxidants, 2019, 8(10): 429. doi: 10.3390/antiox8100429 [26] ZHENG X S, JI H W, ZHANG D, et al. The identification of three phospholipid species roles on the aroma formation of hot-air-dried shrimp (Litopenaeus vannamei) by gas chromatography-ion mobility spectrometry and gas chromatography-mass spectrometry[J]. Food Res Int, 2022, 162(PB): 112191. [27] RAO Z T, WANG S Q, WANG J Q. Protective effects of psoralidin on IL-1β-induced chondrocyte apoptosis.[J]. Mol Med Rep, 2018, 17(2): 3418-3424. [28] 王晓艳, 李伟霞, 张辉, 等. 补骨脂及其主要成分对人正常肝细胞L02的损伤作用研究[J]. 中医研究, 2020, 33(4): 59-63. doi: 10.3969/j.issn.1001-6910.2020.04.24 [29] 刘刚, 冯美云, 张缨, 等. 补充苹果酸复合营养液对拳击运动员抗疲劳能力的影响[J]. 北京体育大学学报, 2009, 32(5): 47-49. [30] LI X Y, ZHENG S X, WU G Y. Nutrition and metabolism of glutamate and glutamine in fish[J]. Amino acids, 2020, 52(5): 671-691. doi: 10.1007/s00726-020-02851-2 [31] JUN J, LONG Y, LI J Y, et al. Glutamate attenuates lipopolysaccharide-induced oxidative damage and mRNA expression changes of tight junction and defensin proteins, inflammatory and apoptosis response signaling molecules in the intestine of fish[J]. Fish Shellfish Immum, 2017, 70: 473-484. doi: 10.1016/j.fsi.2017.09.035 [32] 王珂雯, 徐雷, 徐贞贞, 等. 基于液相色谱-四极杆飞行时间质谱方法分析冰鲜鸡肉代谢标志物[J]. 食品科学, 2021, 42(16): 293-303. doi: 10.7506/spkx1002-6630-20200827-377 [33] WANG Q C, XU Z, AI Q H. Arginine metabolism and its functions in growth, nutrient utilization, and immunonutrition of fish[J]. Anim Nutr, 2021, 7(3): 716-727. doi: 10.1016/j.aninu.2021.03.006 [34] CHENG Z Y, CHEN S Q, AN M L, et al. Effects of replacing fish meal with soybean meal, with or without dietary arginine, on growth performance, immune indices and intestinal morphology of grouper, Epinephelus malabaricus[J]. Aquac Res, 2018, 49(9): 2954-2964. doi: 10.1111/are.13754 [35] WANG B B, PAN X SH, JIA J, et al. Strategy and regulatory mechanisms of glutamate feeding to enhance astaxanthin yield in Xanthophyllomyces dendrorhous[J]. Enzyme Micro Tech, 2019, 125: 45-52. doi: 10.1016/j.enzmictec.2019.02.010 -
下载:
点击查看大图
图(6) / 表(3)
计量
- 文章访问数: 55
- HTML全文浏览量: 41
- PDF下载量: 3
- 被引次数: 0
粤公网安备 44010502001741号
