Synthetic biology approaches and combinatorial biosynthesis towards heterologous lipopeptide production† †Electronic supplementary information (ESI) available: Experimental procedures, design of artificial gene cluster, genetic manipulation, LC-MS analysis and structure elucidation. See DOI: 10.1039/c8sc02046a

Synthetic biology techniques coupled with heterologous secondary metabolite production offer opportunities for the discovery and optimisation of natural products. Synthetic biology techniques coupled with heterologous secondary metabolite production offer opportunities for the discovery and optimisa...

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Published in:Chemical science (Cambridge) 2018-08, Vol.9 (38), p.7510-7519
Main Authors: Yan, Fu, Burgard, Christian, Popoff, Alexander, Zaburannyi, Nestor, Zipf, Gregor, Maier, Josef, Bernauer, Hubert S., Wenzel, Silke C., Müller, Rolf
Format: Article
Language:English
Subjects:
Chemistry
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Summary:Synthetic biology techniques coupled with heterologous secondary metabolite production offer opportunities for the discovery and optimisation of natural products. Synthetic biology techniques coupled with heterologous secondary metabolite production offer opportunities for the discovery and optimisation of natural products. Here we developed a new assembly strategy based on type IIS endonucleases and elaborate synthetic DNA platforms, which could be used to seamlessly assemble and engineer biosynthetic gene clusters (BGCs). By applying this versatile tool, we designed and assembled more than thirty different artificial myxochromide BGCs, each around 30 kb in size, and established heterologous expression platforms using a derivative of Myxococcus xanthus DK1622 as a host. In addition to the five native types of myxochromides (A, B, C, D and S), novel lipopeptide structures were produced by combinatorial exchange of nonribosomal peptide synthetase (NRPS) encoding genes from different myxochromide BGCs. Inspired by the evolutionary diversification of the native myxochromide megasynthetases, the ancestral A-type NRPS was engineered by inactivation, deletion, or duplication of catalytic domains and successfully converted into functional B-, C- and D-type megasynthetases. The constructional design approach applied in this study enables combinatorial engineering of complex synthetic BGCs and has great potential for the exploitation of other natural product biosynthetic pathways.
ISSN:2041-6520
2041-6539
DOI:10.1039/c8sc02046a