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|Title:||Establishing a New Methodology for Genome Mining and Biosynthesis of Polyketides and Peptides through Yeast Molecular Genetics||Authors:||Ishiuchi, K.
Fungal putative gene cluster
Nonribosomal peptide synthetase
Yeast heterologous biosynthesis
|Issue Date:||Apr-2012||Citation:||Ishiuchi, K., Nakazawa, T., Ookuma, T., Sugimoto, S., Sato, M., Tsunematsu, Y., Ishikawa, N., Noguchi, H., Hotta, K., Moriya, H., Watanabe, K. (2012-04). Establishing a New Methodology for Genome Mining and Biosynthesis of Polyketides and Peptides through Yeast Molecular Genetics. ChemBioChem 13 (6) : 846-854. ScholarBank@NUS Repository. https://doi.org/10.1002/cbic.201100798||Abstract:||Fungal genome sequencing has revealed many genes coding for biosynthetic enzymes, including polyketide synthases and nonribosomal peptide synthetases. However, characterizing these enzymes and identifying the compounds they synthesize remains a challenge, whether the genes are expressed in their original hosts or in more tractable heterologous hosts, such as yeast. Here, we developed a streamlined method for isolating biosynthetic genes from fungal sources and producing bioactive molecules in an engineered Saccharomyces cerevisiae host strain. We used overlap extension PCR and yeast homologous recombination to clone desired fungal polyketide synthase or a nonribosomal peptide synthetase genes (5-20 kb) into a yeast expression vector quickly and efficiently. This approach was used successfully to clone five polyketide synthases and one nonribosomal peptide synthetase, from various fungal species. Subsequent detailed chemical characterizations of the resulting natural products identified six polyketide and two nonribosomal peptide products, one of which was a new compound. Our system should facilitate investigating uncharacterized fungal biosynthetic genes, identifying novel natural products, and rationally engineering biosynthetic pathways for the production of enzyme analogues possessing modified bioactivity. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.||Source Title:||ChemBioChem||URI:||http://scholarbank.nus.edu.sg/handle/10635/100603||ISSN:||14394227||DOI:||10.1002/cbic.201100798|
|Appears in Collections:||Staff Publications|
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