Engineering Aspergillus nidulans for heterologous ent-kaurene and gamma-terpinene production.
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.
VTT Technical Research Centre of Finland Ltd., 02044 VTT, Espoo, Finland. kirsi.bromann@helsinki.fi.
Kirsi
K
Bromann
VTT Technical Research Centre of Finland Ltd., 02044 VTT, Espoo, Finland.
Mervi
M
Toivari
Orion Pharma, Orion Corporation, Orionintie 1, 02200, Espoo, Finland.
Kaarina
K
Viljanen
VTT Technical Research Centre of Finland Ltd., 02044 VTT, Espoo, Finland.
Laura
L
Ruohonen
VTT Technical Research Centre of Finland Ltd., 02044 VTT, Espoo, Finland.
Tiina
T
Nakari-Setälä
D019883:Alkyl and Aryl Transferases / Q000235:genetics* / Q000378:metabolism; D001233:Aspergillus nidulans / Q000201:enzymology / Q000235:genetics*; D002957:Citrus / Q000201:enzymology; D000081005:Cyclohexane Monoterpenes; D004271:DNA, Fungal / Q000235:genetics; D045786:Diterpenes, Kaurane / Q000096:biosynthesis*; D005670:Fusarium / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic; D005818:Genetic Engineering; D056426:Genetic Loci; D007218:Industrial Microbiology; D039821:Monoterpenes / Q000378:metabolism*; D005810:Multigene Family; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D000081005:Cyclohexane Monoterpenes; D004271:DNA, Fungal; D045786:Diterpenes, Kaurane; D039821:Monoterpenes; D010940:Plant Proteins; D014157:Transcription Factors; C018669:gamma-terpinene; D019883:Alkyl and Aryl Transferases; C105712:ent-kaurene synthetase B; C021459:kaurene synthetase
Aspergillus nidulans; Gamma-terpinene; Terpenoids; ent-kaurene; ent-pimara-8(14),15-diene