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Improvement of food functions of soybean glycinin by protein engineering and transformation of modified glycine genes into tobacco.

通过蛋白质工程改善大豆甘氨酸的食品功能并将修饰的甘氨酸基因转化到烟草中。

基本信息

批准号：
02660140
负责人：
UTSUMI Shigeru
金额：
$ 1.41万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02660140/
关键词：
Soybean proteins Glycinin Protein engineering Transgenic plant Tobacco

项目摘要

Improvement in nutritional value and functional properties of plant proteins is attracting the attention for their potential in relieving the problem of both starvation and nutritional diseases. In order to establish. fundamental theory to develop novel soybean plants which produce improved proteins, improvement of food functions of soybean glycine by protein engineering and generation of transgenic tobacco plants which are transformed with modified glycine genes were attempted.1. The A_<1a>B_<1b> proglycinin cDNA was modified by insertion of synthetic DNAs encoding four continuous methionines to the cDNA regions corresponding to the variable domains and by deletion of each variable domain or disulfide bonds. The design strategy was based on the structural characteristics of glycine and the relationships between the structure and the functional properties of glycinin. The modified glycines expressed from the modified cDNAs in Escherichia coli were purified, and their abilities to form … More the correct conformation and their functional properties were investigated. Most of the modified glycinins having the abilities to form the correct conformation-exhibited better functional properties than the hative soybean glycinin.2. Investigation on the properties of the modified glycinins with the combination of the modifications described in 1 indicated that the modifications introduced into the glycinin do not induce wrong conformation at all. We can expect that those modified-combined glycinins may exhibit better functional properties than the modified glycinins.3. It was suggested that the yeast expression system of glycinin EDNA is useful for studying protein engineering of glycinin, because yeast can accumulate the modified glycinin containing a modification which disturbs the correct folding.4. The modified glycinin cDNAs encoding the proteins exhibiting high quality of food functions were transformed into tobacco cells using Ti plasmid system and transgenic tobacco plants were regenerated. The modified proteins were accumulated in some tissues of tobacco plants. Less

植物蛋白的营养价值和功能性质的改善因其在缓解饥饿和营养性疾病方面的潜力而引起人们的关注。才能确定。本论文的主要研究内容如下：1.探索了开发生产改良蛋白质的大豆新植株的基础理论，利用蛋白质工程技术改善大豆甘氨酸的食用功能，获得转基因烟草植株。A_&lt；1a&gt；B_&lt；1b&gt；1b&gt；1b&gt；编码四个连续蛋氨酸的合成DNA插入到与可变区对应的cDNA区，并通过删除每个可变区或二硫键来修饰A_&lt；1a&gt；1b&gt；设计策略是基于甘氨酸的结构特征和甘氨酸的结构与功能性质之间的关系。对修饰后的DNA在大肠杆菌中表达的修饰甘氨酸进行了纯化，并对其形成…的能力进行了研究进一步研究了该化合物的正确构象及其功能性质。大多数修饰的大豆球蛋白具有形成正确构象的能力，表现出比天然大豆球蛋白更好的功能特性。结合文献1中所述的修饰，对修饰的球蛋白的性质的研究表明，引入球蛋白的修饰根本不会导致错误构象。我们可以预期，这些修饰结合的球蛋白可能会表现出比修饰的球蛋白更好的功能性质。这表明酵母表达系统可以用于研究大豆球蛋白的蛋白质工程，因为酵母可以积累含有干扰正确折叠的修饰的球蛋白。利用Ti质粒系统将编码高质量食品功能蛋白的修饰后的Glycinin cDNAs转化烟草细胞，获得转基因烟草植株。修饰后的蛋白质在烟草植株的某些组织中积累。较少