The active conformation of avilamycin A is conferred by AviX12, a radical AdoMet enzyme.
The antibiotic avilamycin A is produced by Streptomyces viridochromogenes Tü57. Avilamycin belongs to the family of orthosomycins with a linear heptasaccharide chain linked to a terminal dichloroisoeverninic acid as aglycone. The gene cluster for avilamycin biosynthesis contains 54 open reading frames. Inactivation of one of these genes, namely aviX12, led to the formation of a novel avilamycin derivative named gavibamycin N1. The structure of the new metabolite was confirmed by mass spectrometry (MS) and NMR analysis. It harbors glucose as a component of the heptasaccharide chain instead of a mannose moiety in avilamycin A. Antibacterial activity tests against a spectrum of Gram-positive organisms showed that the new derivative possesses drastically decreased biological activity in comparison to avilamycin A. Thus, AviX12 seems to be implicated in converting avilamycin to its bioactive conformation by catalyzing an unusual epimerization reaction. Sequence comparisons grouped AviX12 in the radical S-adenosylmethionine protein family. AviX12 engineered with a His tag was overexpressed in Escherichia coli and purified by affinity chromatography. The iron sulfur cluster [Fe-S] present in radical AdoMet enzymes was detected in purified AviX12 by means of electron paramagnetic resonance spectroscopy.
Institut für Pharmazeutische Wissenschaften, Pharmazeutische Biologie und Biotechnologie, Albert-Ludwigs-Universität Freiburg, Stefan-Meier-Strasse 19, D-79104 Freiburg, Germany.
Raija
R
Boll
Carsten
C
Hofmann
Björn
B
Heitmann
Gerd
G
Hauser
Steffen
S
Glaser
Thorsten
T
Koslowski
Thorsten
T
Friedrich
Andreas
A
Bechthold
D002236:Carbohydrate Conformation; D004578:Electron Spin Resonance Spectroscopy; D005816:Genetic Complementation Test; D009682:Magnetic Resonance Spectroscopy; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry; D016366:Open Reading Frames; D011134:Polysaccharides / Q000737:chemistry; D018809:Proliferating Cell Nuclear Antigen / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D019998:Racemases and Epimerases / Q000737:chemistry*; D012436:S-Adenosylmethionine / Q000737:chemistry*
D009844:Oligosaccharides; D011134:Polysaccharides; D018809:Proliferating Cell Nuclear Antigen; D012436:S-Adenosylmethionine; D019998:Racemases and Epimerases; C436976:avilamycin A