Identification of the novobiocin biosynthetic gene cluster of Streptomyces spheroides NCIB 11891.
The novobiocin biosynthetic gene cluster from Streptomyces spheroides NCIB 11891 was cloned by using homologous deoxynucleoside diphosphate (dNDP)-glucose 4,6-dehydratase gene fragments as probes. Double-stranded sequencing of 25.6 kb revealed the presence of 23 putative open reading frames (ORFs), including the gene for novobiocin resistance, gyrB(r), and at least 11 further ORFs to which a possible role in novobiocin biosynthesis could be assigned. An insertional inactivation experiment with a dNDP-glucose 4, 6-dehydratase fragment resulted in abolishment of novobiocin production, since biosynthesis of the deoxysugar moiety of novobiocin was blocked. Heterologous expression of a key enzyme of novobiocin biosynthesis, i.e., novobiocic acid synthetase, in Streptomyces lividans TK24 further confirmed the involvement of the analyzed genes in the biosynthesis of the antibiotic.
Steffensky
M
M
Pharmazeutische Biologie, Pharmazeutisches Institut, Eberhard-Karls-Universität Tübingen, 72076 Tübingen, Germany.
Mühlenweg
A
A
Wang
Z X
ZX
Li
S M
SM
Heide
L
L
D019732:Amide Synthases / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003360:Cosmids / Q000235:genetics; D004269:DNA, Bacterial; D015723:Gene Library; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D005810:Multigene Family; D016254:Mutagenesis, Insertional; D009675:Novobiocin / Q000096:biosynthesis* / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism
D004269:DNA, Bacterial; D009675:Novobiocin; D019732:Amide Synthases; C021884:novobiocic acid synthetase