New Synthesis of Polypeptides by Enzyme Systems Cotrolled by Chemical and Physical Methods

化学和物理方法控制的酶系统合成多肽的新方法

• 批准号: 62470103
• 负责人: KUNUGI Shigeru
• 金额: $ 0.96万
• 依托单位: Fukui University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-62470103/
• 关键词: Enzyme; Polypeptides; Cotrol of Function; Chemical Modification; High Pressure; Organic Solvent; 高塩濃度; 高圧力; 高pH; プロテアーゼ; カルボキシペプチダーゼ; サーモライシン; ペプチド合成; 圧力依存性; 加水分解

Polypeptides have wide variety of functions, which primarily depends on the 1st order structure, amino acid sequence. For the ordered polymerization of amino acids utilization of biocatalysts is preferable but, in general, attempts of utilization of bio-specific catalysts such as enzymes are inherently limited by the natural character of the catalysts and often powerless for the preparation of completely new substances.In peptide synthesis usually two types of biocatalysis are used, namely reverse catalysis of proteases, in which variation of peptides applicable for one type of enzyme is limited, and in-vitro protein synthetic system using DNA-mRNA template, which is incapable to the synthesis of non-natural peptides.To overcome these points, this research project aimed to create new enzymatic system by using chemical and biochemical modification of the protein and also by applying highly extreme physical conditions on these enzyme catalytic reactions. That is; thermolysin and serine carboxypeptidases were modified and used for the protein condensation and the reactions were performed under high pressure, high salt concentration or high organic solvent content. Effects of pressure and organic solvent on misactivation by amino acyl-tRNA synthetase was also investigated.

多肽具有多种功能，这主要取决于一级结构、氨基酸序列。对于氨基酸的有序聚合，优选利用生物催化剂，但一般来说，利用生物特异性催化剂（例如酶）的尝试本质上受到催化剂的天然特性的限制，并且通常对于制备全新物质无能为力。在肽合成中，通常使用两种类型的生物催化，即蛋白酶的逆催化，其中 适用于一种酶的肽有限，并且使用DNA-mRNA模板的体外蛋白质合成系统无法合成非天然肽。为了克服这些问题，本研究项目旨在通过对蛋白质进行化学和生化修饰，并在这些酶催化反应上应用高度极端的物理条件来创建新的酶系统。那是;改良嗜热菌蛋白酶和丝氨酸羧肽酶用于蛋白质缩合，反应在高压、高盐浓度或高有机溶剂含量下进行。还研究了压力和有机溶剂对氨酰-tRNA 合成酶失活的影响。

项目成果

Shigeru Kunugi: "Effect of Acylation on Peptide Synthesis by Immobilized Thermolysin" Polymer Journal. 20. 945-947 (1988)

Shigeru Kunugi：“酰化对固定化嗜热菌肽合成的影响”聚合物杂志。

Shigeru Kunugi: "Mechanism of Salt Activation of Thermolysin" Biocatalysis. in press.

Shigeru Kunugi：“嗜热菌蛋白酶的盐激活机制”生物催化。

森川良雄;近藤俊弘;功刀滋;野村哲士: 福井大学工学部繊維・機能性材料研究施設報告. 25. (1987)

Yoshio Morikawa；Toshihiro Kondo；Tetsushi Nomura：福井大学工程学院纤维与功能材料研究设施报告。（1987）

功刀滋;森川良雄;野村哲士;北野博己: 福井大学工学部研究報告. 36. (1988)

Shigeru Koto；Yoshio Morikawa；Tetsushi Nomura；Hiroki Kitano：福井大学工程学院的研究报告。（1988）

Shigeru Kunugi: Bull.Chem.Soc,Japan. 62. 514-518 (1989)

Shigeru Kunugi：Bull.Chem.Soc，日本。