Cloning and analysis of a type II polyketide synthase gene cluster from Streptomyces toxytricini NRRL 15,443 |
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Authors: | Yoo Anna Demirev Atanas V Lee Ji Seon Kim Sang Dal Nam Doo Hyun |
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Institution: | Faculty of Pharmacy, Yeungnam University, 214-1 Dae-dong, Kyongsan 712-749, Republic of Korea. |
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Abstract: | 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. |
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