Gentamicin C2b

Gentamicin C1a and the minor isomer C2b have been reported to have favorable properties in terms of antibacterial activity and toxicity compared to the commercial mixture from which they have previously been isolated by preparative high-performance liquid chromatography. We report straightforward syntheses of both compounds from readily available sisomicin by selective oxidation of the side chain in ring I, hydrogenation of the double bond in ring I to give the 5'-epi series, inversion of configuration at position 5' under thermodynamic conditions, and installation of the 6'-amino group by reductive amination.

Gentamicin C complex fromMicromonospora echinosporaremains a globally important antibiotic, and there is revived interest in the semisynthesis of analogs that might show improved therapeutic properties. The complex consists of five components differing in their methylation pattern at one or more sites in the molecule. We show here, using specific gene deletion and chemical complementation, that the gentamicin pathway up to the branch point is defined by the selectivity of the methyltransferases GenN, GenD1, and GenK. Unexpectedly, they comprise a methylation network in which early intermediates are ectopically modified. Using whole-genome sequence, we have also discovered the terminal 6'-N-methyltransfer required to produce gentamicin C2b from C1a or gentamicin C1 from C2, an example of an essential biosynthetic enzyme being located not in the biosynthetic gene cluster but far removed on the chromosome. These findings fully account for the methylation pattern in gentamicins and open the way to production of individual gentamicins by fermentation, as starting materials for semisynthesis.

A gentamicin C-complex preparation was separated preparatively into five components by Craig distribution. Gentamicins C1, C2, and C1a were the major components, whereas components C2a and C2b represented only 4% of the total C-complex mixture. Paper chromatographic analysis showed that the gentamicin C2b separated by Craig distribution was identical with a gentamicin isolated from Micromonospora purpurea var. JI-33 fermentation broth and identified as 6'-N-methylgentamicin C1a. Similarly, component C2a was identical with a previously separated gentamicin component tentatively identified as the 6'-C epimer of gentamicin C2.