Simultaneous biosynthesis of putrebactin, avaroferrin and bisucaberin by Shewanella putrefaciens and characterisation of complexes with iron(III), molybdenum(VI) or chromium(V)

Cultures of Shewanella putrefaciens grown in medium containing 10 mM 1,4-diamino-2-butanone (DBO) as an inhibitor of ornithine decarboxylase and 10 mM 1,5-diaminopentane (cadaverine) showed the simultaneous biosynthesis of the macrocyclic dihydroxamic adds: putrebactin (pbH(2)), avaroferrin (avH(2)) and bisucaberin (bsH(2)). The level of DBO did not completely repress the production of endogenous 1,4-diaminobutane (putrescine) as the native diamine substrate of pbH(2). The relative concentration of pbH(2):avH(2):bsH(2) was 1:2:1, which correlated with the substrate selection of putrescine:cadaverine in a ratio of 1:1. The macrocycles were characterised using LC-MS as free ligands and as 1:1 complexes with Fe(III) of the form [Fe(pb)](+), [Fe(av)](+) or [Fe(bs)](+), with labile ancillary ligands in six-coordinate complexes displaced during ESI-MS acquisition; or with Mo(VI) of the form [Mo(O)(2)(pb)], [Mo(O)(2)(av)] or [Mo(O)(2)(bs)]. Chromium(V) complexes of the form [CrO(pb)](+) were detected from solutions of Cr(VI) and pbH(2) in DMF using X-band EPR spectroscopy. Supplementation of S. putrefaciens medium with DBO and 1,3-diaminopropane, 1,6-diaminohexane or 1,4-diamino-2(Z)-butene (Z-DBE) resulted only in the biosynthesis of pbH(2). The work has identified a native system for the simultaneous biosynthesis of a suite of three macrocyclic dihydroxamic acid siderophores and highlights both the utility of precursor-directed biosynthesis for expanding the structural diversity of siderophores, and the breadth of their coordination chemistry. (C) 2015 Elsevier Inc. All rights reserved.