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天然タンパク質の加水分解による生理活性ペプチドの新しい抽出合成法

水解天然蛋白质提取合成生物活性肽的新方法

基本信息

批准号：
10650764
负责人：
HIRATA Akira
金额：
$ 2.56万
依托单位：
Waseda University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

项目摘要

We have aimed, in this study, (1)to control the selective production of desired peptides (e.g. angiotensin-converting enzyme (ACE)-inhibitory peptides) using the hydrolysis of corn proteins (zeins) catalyzed by proteases and (2) to develop the enzyme preparation catalytically active in organic media. The following results were obtained :1. We found that zein selectively partitioned to the upper phase, limited hydrolysates to both phases and peptides with low molecular weights selectively to the bottom phase when corn protein α-zein was hydrolyzed in organic polymer two-phase systems. The method proposed in this study can solve the problems of a protein hydrolysis in conventional homogeneous reaction media such as difficulties in the selective synthesis of products and the separation of products and an enzyme.2 We proposed the novel hybrid enzyme preparation catalytically active in organic media, i.e. the noncovalently-formed complex between an enzyme and polymers. It was found that the complex was catalytically active in organic media even when the molar ratio of the polymer/enzyme in the preparation stage was only a unity. The complex showed a high activity when hydrophobic buffers and amphiphilic polymers were used in its preparation stage. PEG4000-α-chymotrypsin complex prepared from a boric acid-succinic acid buffer solution showed ca. 8000-fold higher activity than native α-chymotrypsin in anhydrous isooctane. The method to modify enzymes proposed in this study was novel and had big ripple effects to biology. The complex was catalytically active also in organic two-phase systems and could be applied to the enzymatic hydrolysis of proteins.

在这项研究中，我们的目标是：(1)利用蛋白酶催化玉米蛋白（zeins）的水解来控制所需肽（如血管紧张素转换酶（ACE）抑制肽）的选择性生产；(2)开发在有机培养基中具有催化活性的酶制剂。得到了以下结果：1。研究发现，玉米蛋白α-玉米蛋白在有机聚合物两相体系中水解时，玉米蛋白选择性地向上相分离，将水解产物限制在两相，而低分子量肽选择性地向下相分离。本研究提出的方法解决了在常规均相反应介质中水解蛋白质时，产物的选择性合成困难、产物与酶分离困难等问题我们提出了一种在有机介质中具有催化活性的新型杂化酶制剂，即酶与聚合物之间形成的非共价络合物。在制备阶段，当聚合物/酶的摩尔比为一个单位时，该配合物在有机介质中仍具有催化活性。在制备阶段使用疏水缓冲液和两亲性聚合物时，该配合物表现出较高的活性。硼酸-琥珀酸缓冲液制备的PEG4000-α-胰凝乳酶复合物在无水异辛烷中的活性比天然α-胰凝乳酶高约8000倍。本研究提出的酶修饰方法新颖，在生物学上具有较大的连锁效应。该配合物在有机两相体系中也具有催化活性，可用于蛋白质的酶解。