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[6个高粱品种(双色高粱L. Moench)籽粒醇溶蛋白的提取与特性分析]

[Extraction and characterization of grain prolamins of 6 sorghum Sorghum bicolor L. Moench cultivars].

作者信息

Ortíz de Bertorelli L

机构信息

Instituto de Química y Tecnología, Facultad de Agronomía (UCV), Maracay, Venezuela.

出版信息

Arch Latinoam Nutr. 1992 Mar;42(1):46-51.

PMID:1308645
Abstract

The purpose of this work was to isolate and characterize prolamines from grains of sorghum, by their solubility and molecular properties. One variety, Zaraza-1 (SV-V51), and five hybrids (Dekalb-55, Dekalb-64, Wac-5005, Pioneer-815B and Savanna-5) were used. The prolamines, called kafirins, were extracted with 70% isopropanol at 60 degrees C, and then fractionated by two methods: 95% ethanol and gel filtration with Sephadex G-200. The molecular weight was determined by electrophoresis in a discontinuous polyacrylamide gel with sodium dodecyl sulfate (PAGE-SDS). The alpha kafirin (soluble fraction in 95% ethanol), and 7.13% of the grain's total protein, and the beta kafirin represents 21.64% of kafirin (the insoluble fraction), represents 78.36% of the kafirin and 25.39% of total protein. Three fractions were obtained from the column fractionation, two important ones (A and C), and one (B) in a lesser amount. The proportion of these fractions varied between cultivars, so that in SV-V51, D-64 and Wac-5005, A is predominant, and in high tannin sorghum, C is the predominant one. Kafirins reduced with 2-mercaptoethanol were separated in four subunits with molecular weights of: 15,000, 21,300, 24,300 and 51,500 daltons. Unreduced kafirin showed a larger number of subunits (six) whose molecular weights were: 15,000, 24,300, 48,500, 69,000, 80,000 and 93,500 daltons. Reduced and kafirin showed the same number of subunits as those of nonfractioned kafirin reduced with 2-mercaptoethanol.

摘要

这项工作的目的是通过高粱籽粒醇溶蛋白的溶解性和分子特性对其进行分离和表征。使用了一个品种Zaraza - 1(SV - V51)和五个杂交种(Dekalb - 55、Dekalb - 64、Wac - 5005、Pioneer - 815B和Savanna - 5)。将被称为高粱醇溶蛋白的醇溶蛋白在60℃下用70%异丙醇提取,然后通过两种方法进行分级分离:95%乙醇和用葡聚糖凝胶G - 200进行凝胶过滤。分子量通过在含有十二烷基硫酸钠的不连续聚丙烯酰胺凝胶中进行电泳(PAGE - SDS)来测定。α-高粱醇溶蛋白(在95%乙醇中的可溶部分)占籽粒总蛋白的7.13%,β-高粱醇溶蛋白占高粱醇溶蛋白的21.64%(不溶部分),占高粱醇溶蛋白的78.36%和总蛋白的25.39%。从柱分级分离中获得了三个级分,两个重要级分(A和C),一个含量较少的级分(B)。这些级分的比例在不同品种间有所变化,因此在SV - V51、D - 64和Wac - 5005中,A占主导,而在高单宁高粱中,C占主导。用2 - 巯基乙醇还原后的高粱醇溶蛋白被分离为四个亚基,分子量分别为:15,000、21,300、24,300和51,500道尔顿。未还原的高粱醇溶蛋白显示出更多的亚基(六个),其分子量分别为:15,000、24,300、48,500、69,000、80,000和93,500道尔顿。还原后的高粱醇溶蛋白显示出与用2 - 巯基乙醇还原的未分级高粱醇溶蛋白相同数量的亚基。

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