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An Efficient System for Heterologous Expression of Secondary Metabolite Genes in Aspergillus nidulans

Chiang, Yi-Ming
Oakley, C. Elizabeth
Ahuja, Manmeet
Entwistle, Ruth A.
Schultz, Aric
Chang, Shu-Lin
Wang, Clay C. C.
Oakley, Berl R.
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Abstract
Fungal secondary metabolites (SMs) are an important source of medically valuable compounds. Genome projects have revealed that fungi have many SM biosynthetic gene clusters that are not normally expressed. To access these potentially valuable, cryptic clusters, we have developed a heterologous expression system in Aspergillus nidulans. We have developed an efficient system for amplifying genes from a target fungus, placing them under control of a regulatable promoter, transferring them into A. nidulans and expressing them. We have validated this system by expressing non-reducing polyketide synthases of Aspergillus terreus and additional genes required for compound production and release. We have obtained compound production and release from six of these NR-PKSs and have identified the products. To demonstrate that the procedure allows transfer and expression of entire secondary metabolite biosynthetic pathways, we have expressed all the genes of a silent A. terreus cluster and demonstrate that it produces asperfuranone. Further, by expressing the genes of this pathway in various combinations, we have clarified the asperfuranone biosynthetic pathway. We have also developed procedures for deleting entire A. nidulans SM clusters. This allows us to remove clusters that might interfere with analyses of heterologously expressed genes and to eliminate unwanted toxins.
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This document is the Accepted Manuscript version of a Published Work that appeared in final form in the Journal of the American Chemical Society, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see http://doi.org/10.1021/ja401945a.
Date
2013-05-22
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American Chemical Society
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Chiang, Y.-M., Oakley, C. E., Ahuja, M., Entwistle, R., Schultz, A., Chang, S.-L., … Oakley, B. R. (2013). An Efficient System for Heterologous Expression of Secondary Metabolite Genes in Aspergillus nidulans. Journal of the American Chemical Society, 135(20), 7720–7731. http://doi.org/10.1021/ja401945a
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