To investigate the distribution of the mevalonate pathway within Actinobacteria, we compared a neighbor-joining phylogenetic tree using amino acid sequences of hmgr (Fig. 1a) with a tree calculated from 16S rRNA gene sequences (Fig. 1b). The results showed no specific corelation between the HMGR and the 16S rRNA gene trees. Members of the genus Streptomyces grouped as diverse clades in the HMGR tree and strains belonging to other genera (Actinoplanes, Nocardia, Mycobacterium, and Micromonospora) formed monophyletic clades supported by high bootstrap values (Fig. 1a). In previous studies, it has been reported that this lack of

corelation Ion Channel Ligand Library between established organismal phylogeny and HMGR trees may be attributed to the events of lateral gene transfer (Gophna et al., 2005). Therefore, our

results also indicate that the presence of the mevalonate pathway may not be related to organismal phylogeny based on 16S rRNA gene sequences. The strains possessing hmgr were examined for the production of isoprenoids. Culture extracts of these strains Selleckchem Nutlin 3a were analyzed by LC-MS analysis, and their chemical structures were determined by nuclear magnetic resonance spectral, high-resolution electronspray ionization mass spectral, and UV spectroscopic data. Interestingly, a total of five compounds, including novel compounds JBIR-46, -47, and -48, were detected from the cultures of four strains (Table 3). Strain Sp080513GE-23 (Streptomyces sp.) produced a known isoprenoid, fumaquinone (Fig. 2; Charan et al., 2005), which may be synthesized via the Astemizole mevalonate pathway. SpC080624GE-05 (Micromonospora sp.) produced squalene as a primary metabolite, but squalene has been reported to be synthesized via the MEP pathway in Streptomyces (Fontana et al., 2001). Furthermore, SpC080624SC-11 (Streptomyces setonensis) and SpA080624GE-02 (S. setonensis) produced the three novel isoprenoid compounds JBIR-46, -47, and -48 (Fig. 2), which consist of phenazine chromophores. The structures

of these compounds were determined on the basis of the detailed studies of molecular formulae, UV spectra, and 1H and 13C nuclear magnetic resonance spectra. These detailed structure elucidations will be reported elsewhere. In six strains possessing hmgr, we confirmed that three strains produced isoprenoids as secondary metabolites. Unfortunately, the remaining three strains (Sp080513SC-18, Se080624GE-07, and SpC080624GE-05) did not produce isoprenoids via the mevalonate pathway. This may be due to improper culture conditions, such as the use of an unsuitable production medium. Therefore, we are currently attempting to culture these strains under optimal conditions for the production of corresponding compounds.