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Establishing a new methodology for genome mining and biosynthesis of polyketides and peptides through yeast molecular genetics
- Source :
- Chembiochem : a European journal of chemical biology. 13(6)
- Publication Year :
- 2011
-
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.
- Subjects :
- Saccharomyces cerevisiae Proteins
Saccharomyces cerevisiae
Biochemistry
Genome
chemistry.chemical_compound
Polyketide
Biosynthesis
Nonribosomal peptide
Polyketide synthase
Peptide Synthases
Molecular Biology
Gene
chemistry.chemical_classification
biology
Molecular Structure
Organic Chemistry
biology.organism_classification
Yeast
Biosynthetic Pathways
chemistry
biology.protein
Molecular Medicine
Genome, Fungal
Genetic Engineering
Polyketide Synthases
Subjects
Details
- ISSN :
- 14397633
- Volume :
- 13
- Issue :
- 6
- Database :
- OpenAIRE
- Journal :
- Chembiochem : a European journal of chemical biology
- Accession number :
- edsair.doi.dedup.....b3ab1a935d4d4ff5620045fe92d2de17