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作物杂志,2020, 第5期: 98–102 doi: 10.16035/j.issn.1001-7283.2020.05.015

• 遗传育种·种质资源·生物技术 • 上一篇    下一篇

玉米胚乳中最大量蛋白体的提纯与体外消化

张丛卓1(), 邢丽君2, 李南2, 闫枫2, 田怀东2()   

  1. 1山西农业大学（山西省农业科学院）作物科学研究所,030031,山西太原
    2山西大学生命科学学院植物种质与作物遗传资源实验室,030006,山西太原
  • 收稿日期:2020-01-07 修回日期:2020-02-26 出版日期:2020-10-15 发布日期:2020-10-12
  • 通讯作者: 田怀东
  • 作者简介:张丛卓,主要从事玉米遗传育种研究,E-mail： 170740774@qq.com
  • 基金资助:
    山西省科技创新项目(2014101025);山西省科技基础条件平台建设项目(2011091020);山西省科技攻关项目(20090311002-5)

Purification and In-Vitro Digestion of Maximum PBs in Maize Endosperm

Zhang Congzhuo1(), Xing Lijun2, Li Nan2, Yan Feng2, Tian Huaidong2()   

  1. 1Institute of Crop Sciences, Shanxi Agricultural University (Shanxi Academy of Agricultural Sciences),Taiyuan 030031, Shanxi, China
    2Laboratory of Plant Germplasm and Genetic Resources of Crop,College of Life Science, Shanxi University, Taiyuan 030006, Shanxi, China
  • Received:2020-01-07 Revised:2020-02-26 Online:2020-10-15 Published:2020-10-12
  • Contact: Tian Huaidong

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摘要：

为了揭示玉米（Zea mays L.）种质资源醇溶蛋白营养价值的多样性,本研究优化了具有野生型醇溶蛋白性状的玉米自交系PC130胚乳中蛋白体（PBs）的提纯与体外消化环节,结果显示,基于优化后提纯方法的PBs提纯量比已报道方法的获得量提高了50倍以上;通过分析相关因素对最大量PBs提纯物消化的影响,确定0.5mg/mL胃蛋白酶用量与75min处理时间为胚乳PBs的最活跃体外消化条件。

关键词: 玉米, 最大量蛋白体, 提纯, 体外消化

Abstract:

In order to reveal the diversity of nutritional values of zeins in maize germplasm resources, we optimized the extraction and in-vitro digestion of protein bodies (PBs) from the maize inbred line PC130 with the wild type character of zeins. The results showed that the amount of PBs purified with the optimized purification method was more than 50 times higher than the amount based on the reported methods; 0.5mg/mL pepsin-dosage and 75min treatment-time were determined as the conditions for the most active in-vitro digestion of endosperm PBs, by analyzing the effects of the relative factors on digestion of the purified maximum-PBs-components.

Key words: Zea mays L., Maximum PBs, Purification, In-vitro digestion

图1

使用含不同浓度蔗糖的缓冲液提取的PBs组分的SDS-PAGE图像 M：标记;1~8：PBs组分;TP：全蛋白,下同

图2

使用含不同浓度蔗糖的缓冲液提取的PBs组分的含量变化 不同小写字母表示在0.05%水平上显著,下同

图3

最适蔗糖浓度下提取并经等量加水与不同时间静置析出的PBs组分的SDS-PAGE图像 1~5：PBs组分

图4

最适蔗糖浓度下提取并经等量加水与不同时间静置析出的PBs组分的含量变化

图5

最大量PBs组分经不同浓度胃蛋白酶体外消化后所得残留物的SDS-PAGE图像 1~6：PBs组分

图6

最大量PBs组分经不同浓度胃蛋白酶体外消化后消化率的变化

图7

最大量PBs组分经最适胃蛋白酶用量下不同时间体外消化后所得残留物的SDS-PAGE图像 1~8：PBs组分

图8

最大量PBs组分经最适胃蛋白酶用量下不同时间体外消化后消化率的变化

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