Production of 8'-halogenated and 8'-unsubstituted novobiocin derivatives in genetically engineered streptomyces coelicolor strains.
In the present study, we produced a hybrid antibiotic, carrying a chlorine atom instead of a methyl group at position 8 of the aminocoumarin moiety of novobiocin. This compound was not accessible by conventional gene inactivation/gene expression experiments due to difficulties in the genetic manipulation of the novobiocin producer Streptomyces spheroides. However, the desired compound was obtained after modification of the novobiocin biosynthetic gene cluster by using lambda-Red-mediated recombination in Escherichia coli, followed by integration of the resulting modified cosmid into the phiC31 attachment site of Streptomyces coelicolor and coexpression of the halogenase Clo-hal of clorobiocin biosynthesis. The halogenase BhaA, responsible for chlorination of tyrosyl moieties of the glycopeptide antibiotic balhimycin, was unable to functionally replace the halogenase Clo-hal, suggesting that the two enzymes have different substrate specificities.
Eustáquio
Alessandra S
AS
Pharmazeutische Biologie, Pharmazeutisches Institut, Eberhard-Karls-Universität Tübingen Auf der Morgenstelle 8, 72076 Tübingen, Germany.
Gust
Bertolt
B
Li
Shu-Ming
SM
Pelzer
Stefan
S
Wohlleben
Wolfgang
W
Chater
Keith F
KF
Heide
Lutz
L
D000900:Anti-Bacterial Agents / Q000096:biosynthesis; D020868:Gene Silencing; D005818:Genetic Engineering; D006219:Halogens / Q000378:metabolism*; D005810:Multigene Family; D009675:Novobiocin / Q000031:analogs & derivatives* / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D048372:Streptomyces coelicolor / Q000235:genetics* / Q000378:metabolism
D000900:Anti-Bacterial Agents; D006219:Halogens; C499828:novclobiocin 114; D009675:Novobiocin