流水槽养殖与传统池塘养殖的罗非鱼肌肉差异蛋白组学分析

Differential proteomic analysis of muscle of Oreochromis mossambicus between raceway system aquaculture and traditional aquaculture in ponds

目的利用蛋白组学技术对流水槽养殖与传统池塘养殖的罗非鱼肌肉的差异蛋白进行分析,探讨两种养殖模式罗非鱼肉质差异的分子机制。方法采用纳米液相色谱-串联质谱(Nano liquid chromatography-tandem mass spectrometry,Nano LC-MS/MS)技术手段,鉴定两种养殖模式罗非鱼鱼肉差异蛋白,并进行代谢通路的分析。结果在两种鱼肉中共鉴定到蛋白3684个,其中共检测到差异表达的蛋白为26个。与传统池塘养殖的鱼肉相比,流水槽养殖的鱼肉中磷酸化酶b激酶、硫氧还蛋白还原酶、腺苷酸琥珀酸酶、蛋白酶体抑制剂等蛋白的表达量显著提高;而胞质脂肪酸结合蛋白1/2、Ⅻ型胶原蛋白、线粒体导肽水解酶β等表达量则显著降低。这些差异蛋白与机体内蛋白质、脂肪的累积紧密相关,这可能也是本研究中观察到流水槽养殖罗非鱼鱼肉具有高蛋白、低脂肪的原因之一。生物学功能显示差异蛋白主要分布在转运活性方面,而差异蛋白功能通路的富集结果表明大多数差异蛋白与心肌的收缩、甲状腺激素合成、丙氨酸/天冬氨酸/谷氨酸代谢、细胞内吞作用有关。结论本研究得出的差异蛋白和关键代谢通路与鱼肉中蛋白质、脂肪含量密切相关,为阐明流水槽养殖模式下罗非鱼肉品质的形成机制奠定了基础,为后续贮藏、加工方法选择提供依据。

Objective To analyze the differentially abundant muscle proteins from Oreochromis mossambicus between raceway system aquaculture and traditional aquaculture in ponds by using proteomics,and explore the molecular mechanism of the difference of meat quality between 2 kinds of culture system.Methods Nano liquid chromatography-tandem mass spectrometry(Nano LC-MS/MS)was used to identify the differentially abundant proteins in 2 kinds of culture system,and subsequently the differentially abundant proteins were subjected to pathway analysis.Results A total of 3684 proteins were identified in the two kinds of fish muscle,of which 26 proteins were differentially expressed.Compared with fish meat from traditional pond culture,the expression levels of phosphorylase b kinase,thioredoxin reductase,adenylosuccinase,proteasome inhibitor and other proteins in fish meat cultured in raceway were significantly increased.The expression levels of cytoplasmic fatty acid binding protein 1/2,XII collagen and mitochondrial peptide hydrolaseβwere significantly decreased.These differential proteins were closely related to the accumulation of protein and fat in the body.This may also be one of the reasons why Oreochromis mossambicus fish cultured in raceway system had high protein and low fat.Biological function annotation showed that the differentially abundant proteins were mainly distributed in transport activity.The enrichment results of differentially abundant protein functional pathways showed that most of the differential proteins were related to myocardial contraction,thyroid hormone synthesis,alanine/aspartic acid/glutamic acid metabolism,endocytosis.Conclusion The differentially abundant proteins and key metabolic pathways derived from this study are closely related to the protein and fat contents in fish,which lays the foundation for elucidating the mechanism of Oreochromis mossambicus fish muscle quality formation cultured in raceway system,and provides a basis for subsequent storage and processing method selection.