葡萄籽中水溶性钙结合蛋白的分离和鉴定  

Preparation and identification of water-soluble calcium-binding protein from grape(Vitis vinifera L.) seeds

作　　者：吕晨艳[1] 赵广华[1,2] 

机构地区：[1]中国农业大学食品科学与营养工程学院,北京100083 [2]中国航天员科研训练中心航天营养与食品工程重点实验室,北京100094

出　　处：《农业工程学报》2015年第2期319-323,共5页Transactions of the Chinese Society of Agricultural Engineering

基　　金：国家自然科学基金资助项目(31271826)

摘　　要：为了开发植物源葡萄籽补钙制剂，该研究通过亚细胞定位试验表明，葡萄籽的胚乳中含有大量的钙元素。通过电泳分析发现，葡萄籽的水溶性蛋白包括2种主要成分，其中一种是11 S球蛋白（蛋白质B），也是最主要的钙结合蛋白，另一种是表观分子量为670 kDa的蛋白质A。在蛋白质组成相同的情况下，用传统的碱溶酸沉法来分离葡萄籽蛋白会导致大量的钙流失。但用30%～50%硫酸铵沉淀法得到的蛋白质得率是（22.5＆#177;0.02）g/kg，蛋白质中钙质量分数（3.47%）是碱溶酸沉法（1.11%）的3倍。该研究结果为食品工业中矿物结合蛋白质的分离纯化提供参考。Calcium is an essential nutrient required for critical biological functions such as nerve conduction, muscle contraction, mitosis, blood coagulation, and structural support of the skeleton.Dietary calcium intake is of general interest for human beings, particularly for infants and young children, when growth is accelerated. Milk and milk products as effective calcium supplements are generally accepted by human race with their high bioavailability. However, less consumption of milk in industrialized countries leads to inadequate calcium intake. Therefore, it is important to explore an alternate source for calcium supplement. On the other hand, dried grape seeds are likewise rich in lipids （22.07%）, carbohydrates （12.51%） and proteins （11.94%） （w/w） and grape seeds as by-product during juice production can be an alternative source of protein. Meanwhile, this study demonstrates that grape seeds are rich in calcium （（5.62±0.01） g/kg for embryonic cells and （6.32±0.01） g/kg for intact grape seeds）, which was identified by ICP-AES. The calcium was mainly distributed in the stroma of the amyloplasts and around the starch granules, which was observed under TEM （Transmission Electron Microscope）. Further study indicates that water-soluble protein from grape （Vitis viniferaL.） seeds （WSPG） contained two major components, one of which was 11S globulin-like protein mainly responsible for the binding of calcium in WSPG and the other was a novel protein （Protein A）. The calcium contents of protein isolate from each step were identified by ICP-AES as well. When a traditional alkali extraction and acid precipitation method was used for isolation of WSPG, many binding calcium ions were lost. It is worth noting that the protein composition of grape seed protein obtained by both 30%-50% （NH4）2SO4 sediment and the alkali extraction and acid precipitation method was nearly identical, which consisted of protein A and protein B at a ratio of 2 to 3, but the content of calcium in the gra

关 键 词：钙 蛋白 分离 葡萄籽 硫酸铵沉淀 

分 类 号：TS255.2[轻工技术与工程—农产品加工及贮藏工程]