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KMID : 0880220060440060649
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Journal of Microbiology
2006 Volume.44 No. 6 p.649 ~ p.654
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Cloning and Analysis of a Type II Polyketide Synthase Gene Cluster from Streptomyces toxytricini NRRL 15,443
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Yoo An-Na
Demirev Atanas V.
Lee Ji-Seon
Kim Sang-Dal
Nam Doo-Hyun
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Abstract
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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), β-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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KEYWORD
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Type II polyketide synthase, S. toxytricini, lipstatin, β-ketoacyl synthase, chain length factor, acyl carrier protein, thioesterase
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