Cloning and analysis of a type II polyketide synthase gene cluster from Streptomyces toxytricini NRRL 15,443.
A standard type II polyketide synthase (PKS) gene cluster was isolated while attempting to clone the biosynthetic gene for lipstatin from Streptomyces toxytricini NRRL 15,443. This result was observed using a Southern blot of a PstI-digested S. toxytricini chromosomal DNA library with a 444 bp amplified probe of a ketosynthase (KS) gene fragment. Four open reading frames [thioesterase (TE), beta-ketoacyl systhase (KAS), chain length factor (CLF), and acyl carrier protein (ACP)], were identified through the nucleotide sequence determination and analysis of a 4.5 kb cloned DNA fragment. In order to confirm the involvement of a cloned gene in lipstatin biosynthesis, a gene disruption experiment for the KS gene was performed. However, the resulting gene disruptant did not show any significant difference in lipstatin production when compared to wild-type S. toxytricini. This result suggests that lipstatin may not be synthesized by a type II PKS.
Yoo
Anna
A
Faculty of Pharmacy, Yeungnam University, 214-1 Dae-dong, Kyongsan 712-749, Republic of Korea.
Demirev
Atanas V
AV
Lee
Ji Seon
JS
Kim
Sang Dal
SD
Nam
Doo Hyun
DH
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D003470:Culture Media; D017353:Gene Deletion; D007783:Lactones / Q000737:chemistry / Q000378:metabolism; D005810:Multigene Family; D048630:Polyketide Synthases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development
D001426:Bacterial Proteins; D003470:Culture Media; D007783:Lactones; D048630:Polyketide Synthases; C054164:lipstatin