The novel hydroxylation reaction of nicotinic acid catalyzed by the cooperation of yeat and bacteria

各种吡啶衍生物可用作合成各种生理活性化合物的结构单元。例如，最近开发了一种新的强效杀虫剂，即烟碱类的吡虫啉。烟碱类杀虫剂因其活性强、毒性低而被广泛应用。然而，通过常规的化学合成方法合成6-氯吡啶基的结构单元并不容易。因此，需要开发用于合成结构单元的新工艺。为了实现这一目的，我们研究了酶促吡啶环的区域特异性羟基化反应，建立了3-氰基吡啶6位特异性羟基化的新生产工艺。其催化烟酸的2-位特异性羟基化。2-羟基烟酸也可用作合成生理活性化合物的结构单元。 关于我们 化合物.然而，通过使用烟酸作为碳源和/或氮源的常规富集培养，我们不能分离催化烟酸2-位特异性羟基化的微生物。所有烟酸降解微生物都催化烟酸的6位特异性羟基化。因此，我们尝试了使用6-甲基烟酸作为唯一碳源和/或氮源的富集培养。经过长时间的富集培养，发现混合菌具有6-甲基烟酸2-位羟基化活性。混合培养物还催化烟酸的2-位特异性羟基化。从混合培养液中分离到假丝酵母菌和鞘氨醇单胞菌两种微生物。每种菌株都不催化烟酸的羟基化。只有当它们共同培养时，才能发现烟酸的2位特异性羟基化，从而发现了假丝酵母和鞘氨醇单胞菌的协同作用。然而，协同羟基化反应的机理目前尚不清楚。少

Various pyridine derivatives are useful as the building block for the synthesis of various physiologically active compounds. For example, recently, the new powerful insecticide, imidachloprid belonging to nicotinly group, has been developed. These nicotinly insecticides are widely used due to its powerful activity and low toxicity. However, it was not easy to synthesize the building block of 6-chloropyridyl through the conventional chemical synthesis. Thus, the development of the new process for the synthesis of the building block was required. To accomplish this purpose, we investigated the enzymatic regio-specific hydroxylation reaction of pyridine ring and we established the new production process of the 6-position specific hydroxylation of 3-cyanopyridine.Next, we attempted to isolate the microorganisms. which catalyze the 2-position specific hydroxylation of nicotinic acid. 2-Hydroxynicotinic acid might be also useful as the building block for the synthesis of phisiological active … More compounds. However, through the conventional enrichment culture using nicotinic acid as carbon and/or nitrogen source, we could not isolate the microorganisms which catalyze the 2-position specific hydroxylation of nicotinic acid. All nicotinic acid-degrading microorganism catalyzes 6-position specific hydroxylation of nicotinic acid. Thus, we attempted the enrichment culture using 6-methylnicotinic acid as sole carbon and/or nitrogen source. After the prolong enrichment culture, we found the mixed culture exhibit the 2-position hydroxylating activity of 6-methylnicotinic acid. The mixed culture also catalyzed the 2-position specific hydroxylation of nicotinic acid. We found two kinds of microorganism, Candida yeast and Sphingomonas bacteria from the mixed culture broth. Each strain does not catalyze the hydroxylation of nicotinic acid. Only when they were cultivated together, 2-position specific hydroxylation of nicotinic acid was found. Thus, we found the cooperative function of Candida yeast and Sphingomonas bacteria. However, the mechanisms of cooperative hydroxylation are not clear now. Less