|Budget Amount *help
¥3,300,000 (Direct Cost : ¥3,300,000)
Fiscal Year 1998 : ¥500,000 (Direct Cost : ¥500,000)
Fiscal Year 1997 : ¥2,800,000 (Direct Cost : ¥2,800,000)
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
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