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可可豆蛋白质组的纳升超高效液相色谱-电喷雾串联质谱分析。

Characterization of the Proteome of Theobroma cacao Beans by Nano-UHPLC-ESI MS/MS.

机构信息

Department of Chemistry, University of Reading, Reading, UK.

Mondelēz International, Reading Science Centre, Reading, UK.

出版信息

Proteomics. 2018 Feb;18(3-4). doi: 10.1002/pmic.201700339. Epub 2018 Feb 2.

DOI:10.1002/pmic.201700339
PMID:29280563
Abstract

Cocoa seed storage proteins play an important role in flavour development as aroma precursors are formed from their degradation during fermentation. Major proteins in the beans of Theobroma cacao are the storage proteins belonging to the vicilin and albumin classes. Although both these classes of proteins have been extensively characterized, there is still limited information on the expression and abundance of other proteins present in cocoa beans. This work is the first attempt to characterize the whole cocoa bean proteome by nano-UHPLC-ESI MS/MS analysis using tryptic digests of cocoa bean protein extracts. The results of this analysis show that >1000 proteins could be identified using a species-specific Theobroma cacao database. The majority of the identified proteins were involved with metabolism and energy. Additionally, a significant number of the identified proteins were linked to protein synthesis and processing. Several proteins were also involved with plant response to stress conditions and defence. Albumin and vicilin storage proteins showed the highest intensity values among all detected proteins, although only seven entries were identified as storage proteins. A comparison of MS/MS data searches carried out against larger non-specific databases confirmed that using a species-specific database can increase the number of identified proteins, and at the same time reduce the number of false positives. The results of this work will be useful in developing tools that can allow the comparison of the proteomic profile of cocoa beans from different genotypes and geographic origins. Data are available via ProteomeXchange with identifier PXD005586.

摘要

可可豆贮藏蛋白在风味发展中起着重要作用,因为在发酵过程中,它们的降解会形成香气前体。可可豆中的主要蛋白质是属于豆球蛋白和清蛋白类的贮藏蛋白。尽管这两类蛋白质都得到了广泛的研究,但关于可可豆中其他蛋白质的表达和丰度的信息仍然有限。本研究首次尝试通过使用可可豆蛋白提取物的胰蛋白酶消化物进行纳升超高效液相色谱-电喷雾串联质谱(nano-UHPLC-ESI MS/MS)分析来描绘整个可可豆蛋白质组。该分析的结果表明,使用种特异性可可豆数据库可以鉴定出 >1000 种蛋白质。鉴定出的大多数蛋白质与代谢和能量有关。此外,相当数量的鉴定出的蛋白质与蛋白质合成和加工有关。一些蛋白质还参与植物对胁迫条件和防御的反应。清蛋白和豆球蛋白贮藏蛋白在所有检测到的蛋白质中表现出最高的强度值,尽管仅鉴定出 7 种贮藏蛋白。对更大的非特异性数据库进行的 MS/MS 数据搜索比较证实,使用种特异性数据库可以增加鉴定出的蛋白质数量,同时减少假阳性的数量。这项工作的结果将有助于开发工具,允许比较来自不同基因型和地理起源的可可豆的蛋白质组图谱。数据可通过 ProteomeXchange 获得,标识符为 PXD005586。

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