Control of the Functionality by selective modification on protein surface with calixarene compounds

本年度以杯芳烃类化合物为催化剂，利用不溶于有机介质的细胞色素c，研究了芳烃类化合物的氧化聚合（酶催化聚合）。研究了邻苯二胺的氧化聚合反应，确定了反应速率与细胞色素c浓度、氧化化合物的加入量和一种有机溶剂有关。在适当的条件下反应数小时后，形成了分子量为几千的聚合物，但尚未形成较大的聚合物。其次，研究了蛋白质的转运，使某些蛋白质连续分离。在正向和反向提取蛋白质时，这两个速率都取决于一些因素，如萃取剂浓度、饲料和回收阶段的pH值、盐浓度和添加的酒精浓度。基于上述因素的优化体系实现了以杯[6]芳烃羧酸为载体的液膜提取存在于进料相的细胞色素c，并连续剥离至回收相。接下来，研究了二胺的提取，以阐明钙[6]芳烃羧酸对蛋白质的识别。首次合成了具有两个氨基的二肽化合物。对该化合物进行了提取。虽然杯芳烃化合物对二肽化合物具有较高的萃取性能，但建议用单分子杯芳烃化合物进行萃取，即不形成1:2的络合物。最后，制备了新的主体化合物，并对其提取性能进行了研究。该化合物在一个杯形芳烃分子中含有两个吡啶和两个羧基。发现它对银有选择性。还制备了带有烯丙基的杯状[4]芳烃羧酸，用于研究银的提取。

In this year, oxidative polymerization of aromatic compounds (polyrnerizatio with enzyme) has been investigated by using cytochrome c, which is aoluble to organic media with calixarene compounds as catalyst. Oxidative polymerization of o-phenylene diamine has been investigated and the reaction rate is confirmed to be dependent on cytchrome c concentration, added amount of oxidative compound and a kind of organic solvents. After some hours of the reaction under the adequate condition, polymer with a few thousand of molecular weight was formed, but larger polymer has not been formed.Second, transport of protein has been investigated to continuously separate of some proteins. At the forward and back extraction of proteins, both rates are dependent on some factors, such as extractant concentration, pH values of feed and recovery phases, salt concentration, and added alcohol concentration. The optimal system based on the above factors realize that cytchrome c exsisting in feed phase is extracted with liquid membrane containing calix[6]arene carboxyric acid as a carrier and continuously stripped to recovery phase.Next, the extraction of diamine has been investigated for the elucidation of protein recognition with calk[6]arene carboxylic acid. The dipeptide compound with two amino groups was firstly prepared. The extraction of the compound was carried out. Although calyix[6]arene compound showed high extraction property to the dipeptide compound, the guest compound is suggested to be extracted with a single molecule of calixarene compound which means not to form 1:2 complex.Finally, the novel host compound has been prepared to investigate the extraction property. The compound possesses two pyridil and two carboxyl groups in a single molecule of calix[4]arene. It was found to show silver selectivity. Calix[4]arene carboxylic acid with allyl groups has been also prepared to investigate silver extraction.