Landry J
Biochimie. 1979;61(4):549-58. doi: 10.1016/s0300-9084(79)80211-4.
A laboratory procedure for isolation and purification of zein from grains of 4 varieties of Maize was described. The preparations were characterized by their physicochemical properties. Upon polyacrylamide gel electrophoresis in sodium dodecyl sulphate (SDS), native zein (from INRA 260 hybrid) was resolved into 2 major classes with average molecular weights of 45,000 and 22,000. After reduction with mercaptoethanol zein contained only two subunits of 22,000 and 24,000 daltons. Upon starch gel electrophoresis in 6 M urea at pH 3.5, native zein exhibited five major or medium intensity bands and several minor ones. The latter, under reducing conditions, disappeared to reinforce the major bands or to yield some new minor bands. Amino acid analysis revealed a very low content of lysine. The NH2-terminal amino acids were determined to be threonine and phenylalanine with a preponderance of the former. Zeins isolated from the varieties studied appeared tohave the same NH2-terminal residues and similar amino acid compositions with an arginine/histidine ratio ranging from 1.1 to 1.2. They differed in relative importance of components, detected by electrophoresis in the presence of SDS or urea. Changes in zein characteristics with the grain genotype allow one to conclude that the components of molecular weights of 22,000 and 24,000 consist of several subunits differing in charge and amino acid content.
描述了一种从4个玉米品种的谷粒中分离和纯化玉米醇溶蛋白的实验室方法。对制备物的理化性质进行了表征。在十二烷基硫酸钠(SDS)存在下进行聚丙烯酰胺凝胶电泳时,天然玉米醇溶蛋白(来自法国国家农业研究院260杂交种)可分离为两大类,平均分子量分别为45,000和22,000。用巯基乙醇还原后,玉米醇溶蛋白仅含有两个分子量分别为22,000和24,000道尔顿的亚基。在pH 3.5的6 M尿素中进行淀粉凝胶电泳时,天然玉米醇溶蛋白呈现出五条主要或中等强度的条带以及几条次要条带。在还原条件下,后者消失,从而增强了主要条带或产生了一些新的次要条带。氨基酸分析显示赖氨酸含量极低。测定其NH2末端氨基酸为苏氨酸和苯丙氨酸,前者占优势。从所研究的品种中分离出的玉米醇溶蛋白似乎具有相同的NH2末端残基和相似的氨基酸组成,精氨酸/组氨酸比值在1.1至1.2之间。它们在SDS或尿素存在下通过电泳检测的组分相对重要性方面存在差异。玉米醇溶蛋白特性随谷粒基因型的变化使人们可以得出结论,分子量为22,000和24,000的组分由几个电荷和氨基酸含量不同的亚基组成。