Biosynthesis of the terpene phenalinolactone in Streptomyces sp. Tü6071: analysis of the gene cluster and generation of derivatives.
Phenalinolactones are terpene glycosides with antibacterial activity. A striking structural feature is a highly oxidized gamma-butyrolactone of elusive biosynthetic origin. To investigate the genetic basis of the phenalinolactones biosynthesis, we cloned and sequenced the corresponding gene cluster from the producer strain Streptomyces sp. Tü6071. Spanning a 42 kbp region, 35 candidate genes could be assigned to putatively encode biosynthetic, regulatory, and resistance-conferring functions. Targeted gene inactivations were carried out to specifically manipulate the phenalinolactones pathway. The inactivation of a sugar methyltransferase gene and a cytochrome P450 monoxygenase gene led to the production of modified phenalinolactone derivatives. The inactivation of a Fe(II)/alpha-ketoglutarate-dependent dioxygenase gene disrupted the biosynthetic pathway within gamma-butyrolactone formation. The structure elucidation of the accumulating intermediate indicated that pyruvate is the biosynthetic precursor of the gamma butyrolactone moiety.
Dürr
Clemens
C
Albert-Ludwigs-Universität, Institut für Pharmazeutische Wissenschaften, Stefan-Meier-Strasse 19, 79104 Freiburg, Germany.
Schnell
Hans-Jörg
HJ
Luzhetskyy
Andriy
A
Murillo
Renato
R
Weber
Monika
M
Welzel
Katrin
K
Vente
Andreas
A
Bechthold
Andreas
A
D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D006027:Glycosides / Q000096:biosynthesis / Q000737:chemistry; D006601:Hexoses / Q000096:biosynthesis / Q000378:metabolism; D007783:Lactones / Q000737:chemistry / Q000378:metabolism; D008954:Models, Biological; D008969:Molecular Sequence Data; D005810:Multigene Family; D009154:Mutation; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism; D011758:Pyrroles / Q000378:metabolism; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism*; D013729:Terpenes / Q000737:chemistry / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D006027:Glycosides; D006601:Hexoses; D007783:Lactones; D011758:Pyrroles; D013729:Terpenes; C006487:amicetose; D010105:Oxygenases