Identification and quantification of proteins at adsorption layer of emulsion stabilized by pea protein isolates
Identification and quantification of proteins at adsorption layer of emulsion stabilized by pea protein isolates
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豌豆分离蛋白稳定乳液吸附层蛋白质的鉴定和定量
DOI：
10.1016/j.colsurfb.2018.05.068
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Chen Yijie
中科院分区：
文献类型：
--
作者：
Xiong Ting;Ye Xiangdong;Su YanTing;Chen Xi;Sun Hui;Li Bin;Chen Yijie
本研究采用定量蛋白质组学方法对豌豆分离蛋白(PPI)稳定的水包油乳状液中的吸附蛋白质进行了分析。用一种优化的沉淀法沉淀吸附的蛋白质，并对沉淀物进行标记，然后用反相高效液相色谱-串联质谱仪(RPLC-ESI-MS/MS)进行蛋白质鉴定和定量。共鉴定出77个蛋白质，其中有49个蛋白质有显著差异。有25个上调蛋白(Fold Change>； 1)和24个下调蛋白(Fold Change<； 1)。根据蛋白质家族的分类比较了这些蛋白质的界面吸附能力。结果表明，各异构体在油水界面上均表现出较强的吸附能力。与维西林相比，阿罗西林的吸附能力相反。不同豆蛋白家族在油水界面上表现出明显不同的亲和力。与白蛋白-1相反，白蛋白-2优先吸附在界面上。氨基酸序列比对和亲水性分析表明，疏水和亲水氨基酸基团之间平衡良好的蛋白质具有较高的界面活性。相反，较长的亲水或疏水片段可能会对蛋白质界面活性产生不利影响。这项研究通过提供详细的界面层定量信息，对蛋白质的界面行为提供了深入的了解。
This study adopted the method of quantitative proteomics to analyze the adsorbed proteins in oil-in-water emulsions stabilized by pea protein isolate (PPI). Adsorbed proteins were precipitated by an optimized precipitation method and precipitates were labeled and subjected to a reversed-phase high performance liquid chromatography coupled to tandem mass spectrometry (RPLC-ESI–MS/MS) for protein identification and quantification. In total, 77 proteins were identified, of which 49 proteins with significant differences were observed. There were 25 upregulated proteins (fold change > 1) and 24 downregulated proteins (fold change < 1). The interfacial adsorption abilities of these proteins were compared according to the classification of protein families. The results showed that all isoforms of vicilins exhibited high adsorption abilities at the oil-water interface. Compared with vicilin, convicilin showed opposite adsorption capacity. Different legumin families showed significantly different affinities on the oil-water interface. In contrast to albumin-1, albumin-2 was preferentially adsorbed to the interface. The amino acid sequence alignment and hydropathy profile analysis of these proteins showed that the proteins well-balanced between hydrophobic and hydrophilic amino acid groups displayed high interfacial activity. In contrast, a long hydrophilic or hydrophobic fragment could adversely influence protein interfacial activity. This study provides an insight into the interfacial behaviors of proteins by supplying detailed quantitative information of interfacial layer.