A Single Module Type I Polyketide Synthase Directs de Novo Macrolactone Biogenesis during Galbonolide Biosynthesis in Streptomyces galbus

作者：Kim Hyun Ju; Karki Suman; Kwon So Yeon; Park Si Hyung; Nahm Baek Hie; Kim Yeon Ki; Kwon Hyung Jin^*

来源：JOURNAL OF BIOLOGICAL CHEMISTRY, 2014, 289(50): 34557-34568.

DOI：10.1074/jbc.M114.602334

• Actinobacteria
• Bacterial Genetics
• Gene Knockout
• Natural Product Biosynthesis
• Polyketide
• Streptomyces galbus
• Galbonolide
• Highly Reducing Type I Polyketide Synthase

Background: Galbonolide (GAL) was proposed to be synthesized by a modular type I polyketide synthase (PKS) with GalA-E serving a supporting role. Results: GalA-C constitute a sole type I PKS that is involved in GAL biosynthesis in S. galbus. Conclusion: GalA-C catalyze the de novo formation of GAL macrolactone. Significance: GalA-C constitute a novel iterative PKS that incorporates methylmalonate units with highly programmed -keto group modifications.
Galbonolide (GAL) A and B are antifungal macrolactone polyketides produced by Streptomyces galbus. During their polyketide chain assembly, GAL-A and -B incorporate methoxymalonate and methylmalonate, respectively, in the fourth chain extension step. The methoxymalonyl-acyl carrier protein biosynthesis locus (galG to K) is specifically involved in GAL-A biosynthesis, and this locus is neighbored by a gene cluster composed of galA-E. GalA-C constitute a single module, highly reducing type I polyketide synthase (PKS). GalD and GalE are cytochrome P450 and Rieske domain protein, respectively. Gene knock-out experiments verified that galB, -C, and -D are essential for GAL biosynthesis. A galD mutant accumulated a GAL-C that lacked two hydroxyl groups and a double bond when compared with GAL-B. A [U-C-13]propionate feeding experiment indicated that no rare precursor other than methoxymalonate was incorporated during GAL biogenesis. A search of the S. galbus genome for a modular type I PKS system, the type that was expected to direct GAL biosynthesis, resulted in the identification of only one modular type I PKS gene cluster. Homology analysis indicated that this PKS gene cluster is the locus for vicenistatin biosynthesis. This cluster was previously reported in Streptomyces halstedii. A gene deletion of the vinP2 ortholog clearly demonstrated that this modular type I PKS system is not involved in GAL biosynthesis. Therefore, we propose that GalA-C direct macrolactone polyketide formation for GAL. Our studies provide a glimpse into a novel biochemical strategy used for polyketide synthesis; that is, the iterative assembly of propionates with highly programmed -keto group modifications.

• 出版日期2014-12-12