Menaquinone biosyntheses in microorganisms.

In prokaryotes, menaquinone (MK) is involved in an electron-transfer pathway. Its biosynthesis was established in the 1970s and 1980s with Escherichia coli. However, a bioinformatic analysis of whole genome sequences has suggested the presence of an alternative pathway. We investigated a novel pathway in a Streptomyces strain. The (13)C-labeling pattern of MK purified from a Streptomyces strain grown on [U-(13)C]glucose was quite different from that of E. coli. We searched for candidate genes participating in the pathway by in silico screening, and the involvement of these genes in the pathway was confirmed by gene-disruption experiments. We also employed mutagenesis to isolate auxotrophic mutants and used these mutants as hosts for shotgun cloning experiments. Metabolites that accumulated in the culture broth of the mutants were isolated and their structures were determined. Taken together, the results indicated an alternative pathway (futalosine (FL) pathway). Moreover, there were three possible routes in the early part of the FL pathway. FL was directly formed by MqnA in Thermus thermophilus and converted into dehypoxanthinyl FL (DHFL). In Acidothermus cellulolyticus, Streptomyces coelicolor, and Helicobacter pylori, aminodeoxyfutalosine (AFL) was formed by MqnA. In the case of the former two strains, AFL was converted to FL by deaminases then to DHFL. In contrast, AFL was directly converted to DHFL in H. pylori.