Physicochemical properties of native adzuki bean (Vigna angularis) 7S globulin and the molecular cloning of its cDNA isoforms
Physicochemical properties of native adzuki bean (Vigna angularis) 7S globulin and the molecular cloning of its cDNA isoforms
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DOI：
10.1021/jf063205l
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发表时间：
2007-05-02
影响因子：
6.1
通讯作者：
Utsumi, Shigeru
中科院分区：
文献类型：
--
作者：
Fukuda, Takako;Prak, Krisna;Utsumi, Shigeru
7S球蛋白（vicilin）是红小豆[Vigna angularis]中主要的种子贮藏蛋白，通过硫酸铵分级分离、凝胶过滤柱层析和阴离子交换柱层析得到两个组分。在SDS-PAGE上，两种组分都给出了两条主要带和一些次要带，但两种组分之间的次要带组成存在差异。对三种样品的热稳定性、溶解性、表面疏水性和乳化性进行了分析。虽然样品之间的溶解性和乳化能力没有差异，但热稳定性和表面疏水性不同。这些差异可能是由于亚基组成的差异。通过逆转录-聚合酶链反应（RT-PCR），使用根据确定的主要条带的N-末端序列设计的引物克隆cDNA。在豆科植物中，与绿豆8 S α球蛋白（7S球蛋白）同源性最高，其次是大豆β-伴大豆球蛋白（7S球蛋白）β亚基。红小豆7S球蛋白亚型比绿豆8 S α球蛋白和大豆β-伴大豆球蛋白β亚基含有更多的蛋氨酸和色氨酸。此外，高甘露糖型聚糖连接到两个或一个N-糖基化位点的红小豆7S球蛋白。
7S globulin (vicilin), the major seed storage protein in adzuki bean [Vigna angularis], was purified by ammonium sulfate fractionation, gel filtration column chromatography, and anion-exchange column chromatography that resulted in two fractions. On SDS-PAGE, both fractions gave two major and some minor bands, but there was a difference in the minor band compositions between the two fractions. Thermal stability, solubility, surface hydrophobicity, and emulsifying ability of these three samples were analyzed. Although there was no difference in solubility and emulsifying ability among the samples, thermal stability and surface hydrophobicity were different. These differences might be due to the differences in subunit compositions. cDNAs were cloned by reverse transcription-polymerase chain reaction (RT-PCR) using primers designed on the basis of the determined N-terminal sequences of the major bands. We obtained three isoforms of cDNAs, which had highest homology with the mung bean 8S alpha globulin (7S globulin), and then soybean beta-conglycinin (7S globulin) beta subunit among legume plants. Adzuki bean 7S globulin isoforms contain more methionine and tryptophan than mung bean 8S alpha globulin and soybean beta-conglycinin beta subunit. In addition, high mannose types of glycans were attached to two or one N-glycosylation sites of adzuki bean 7S globulins.