Nature Biotechnology
23, 1171 - 1176 (2005)
Published online: 14 August 2005; | doi:10.1038/nbt1128
Combinatorial polyketide biosynthesis by de novo design and rearrangement of modular polyketide synthase genesHugo G Menzella1, 3, Ralph Reid1, 3, John R Carney1, Sunil S Chandran1, Sarah J Reisinger1, Kedar G Patel1, David A Hopwood1, 2
& Daniel V Santi11
Kosan Biosciences, Inc., 3832 Bay Center Place, Hayward, California 94545, USA. 2
Permanent address: Department of Molecular Microbiology, John Innes Centre, Norwich NR4 7UH, UK. 3
These authors contributed equally to this study.
Correspondence should be addressed to Daniel V Santi santi@kosan.com Type I polyketide synthase (PKS) genes consist of modules  3−6 kb long, which encode the structures of 2-carbon units in polyketide products. Alteration or replacement of individual PKS modules can lead to the biosynthesis of 'unnatural' natural products but existing techniques for this are time consuming. Here we describe a generic approach to the design of synthetic PKS genes where facile cassette assembly and interchange of modules and domains are facilitated by a repeated set of flanking restriction sites. To test the feasibility of this approach, we synthesized 14 modules from eight PKS clusters and associated them in 154 bimodular combinations spanning over 1.5-million bp of novel PKS gene sequences. Nearly half the combinations successfully mediated the biosynthesis of a polyketide in Escherichia coli, and all individual modules participated in productive bimodular combinations. This work provides a truly combinatorial approach for the production of polyketides.
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