大豆籽粒贮藏蛋白11S和7S组分提取分离方法的优化  

Optimization of extraction and isolation of 11S and 7S fractions from soybean storage protein

作　　者：刘春[1] 王红玲[1] 崔竹梅[1] 何小玲[2] 王显生[3] 麻浩[1] 

机构地区：[1]南京农业大学国家大豆改良中心 [2]南京农业大学农学院,南京市210095 [3]华南理工大学轻工与食品学院植物蛋白工程研究中心,广州市510640

出　　处：《中国油脂》2006年第2期31-36,共6页China Oils and Fats

基　　金：农业部"948"项目(M2001-207);科技部农业科技成果转化资金项目(02EFN216901241);江苏省"十五攻关"项目(Q200126)

摘　　要：为确定提取分离大豆贮藏蛋白11S和7S两种主要成分的最适方法，采用凯氏定氮和SDS-PAGE分析，从浸提液种类、提取液pH、浸提次数和温度、料液比、Tris-HCl浓度和还原剂种类等影响提取分离效果的因素着手，对Nagano法进一步优化。结果表明，浸提液采用pH8．5、含0．01mol／L亚硫酸氢钠的0．03～0．06mol／L Tris-HCl缓冲液系统，提取温度45℃，料液比1：15，重复浸提两次；分离过程中，在pH6．4沉淀离心分离出11S组分、调pH5．5沉淀离心分离出中间产物后。再调pH至4．8沉淀离心分离出7S组分。优化后的方法与Nagano法相比，可显著提高11S和7S组分的得率、蛋白含量和纯度。In order to determine the optimum conditions for extraction and isolation of 11S and 7S fractions from soybean storage protein, the factors affecting the extraction and isolation（the kinds of extraction solution, alkaline extraction pH,times of extraction and temperature, meal - to- water ratio, Tris- HCl concentration, the kinds of reducing agent） were investigated by Kjeldahl method and SDS - PAGE＇and further more Nagano method was modified. The results indicated that the defatted soybean meal was extracted with 0.03 -0.06 mol/L Tris- HC1 buffer （pH 8.5）containing 0.01 mol/L sodium bisulfite（SBS）at 45℃ for 1 hour.The extraction was repeated twice with meal - to - water ratio 1：15 .The 11S fraction was precipitated at pH 6.4 and isolated by centrifugation, and then the pH of the supernatant was adjusted to pH 5.5 to remove the insoluble intermediate fraction by centrifugation. After then, the supematant was adjusted to pH 4.8 to precipitate and isolate the 7S fraction by centrifugation. Under these conditions, the yield, protein content and purity of llS and 7S fractions could be increased significantly than that by Nagano method.

关 键 词：大豆 贮藏蛋白 11S和7S组分 Nagano法 优化 

分 类 号：TQ936[化学工程]