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Engineering Aspergillus nidulans for heterologous ent-kaurene and gamma-terpinene production
- Bromann, Kirsi, Toivari, Mervi, Viljanen, Kaarina, Ruohonen, Laura, Nakari-Setälä, Tiina
- Applied microbiology and biotechnology 2016 v.100 no.14 pp. 6345-6359
- Aspergillus nidulans, Citrus unshiu, Fusarium fujikuroi, bioactive properties, diterpenoids, engineering, fungi, gamma-terpinene, gene overexpression, genetic engineering, genetically modified organisms, monoterpenoids, multigene family, secondary metabolites, transcription factors
- Terpenes are a large and varied group of natural products with a wide array of bioactivities and applications. The chemical production of industrially relevant terpenes can be expensive and time-consuming due to the structural complexity of these compounds. Here, we studied Aspergillus nidulans as a heterologous host for monoterpene and diterpene production. Previously, we identified a novel diterpene gene cluster in A. nidulans and showed that overexpression of the cluster-specific transcription factor (pbcR) led to ent-pimara-8(14),15-diene (PD) production. We report further characterization of the A. nidulans PD synthase gene (pbcA). In A. nidulans, overexpression of pbcA resulted in PD production, while deletion of pbcA abolished PD production. Overexpression of Fusarium fujikuroi ent-kaurene synthase (cps/ks) and Citrus unshiu gamma-terpinene synthase resulted in ent-kaurene and gamma-terpinene production, respectively. A. nidulans is a fungal model organism and a close relative to other industrially relevant Aspergillus species. A. nidulans is a known producer of many secondary metabolites, but its ability to produce heterologous monoterpene and diterpene compounds has not been characterized. Here, we show that A. nidulans is capable of heterologous terpene production and thus has potential as a production host for industrially relevant compounds. The genetic engineering principles reported here could also be applied to other Aspergilli.