VTT Technical Research Centre of Finland Ltd., 02044 VTT, Espoo, Finland.
Orion Pharma, Orion Corporation, Orionintie 1, 02200, Espoo, Finland.
Appl Microbiol Biotechnol. 2016 Jul;100(14):6345-6359. doi: 10.1007/s00253-016-7517-5. Epub 2016 Apr 20.
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.
萜类化合物是一大类具有广泛生物活性和应用的天然产物，结构多样。由于这些化合物结构复杂，工业相关萜类化合物的化学合成既昂贵又耗时。在这里，我们研究了构巢曲霉作为单萜和二萜生产的异源宿主。之前，我们在构巢曲霉中鉴定了一个新的二萜基因簇，并表明该簇特异性转录因子（pbcR）的过表达导致了-ent-pimara-8(14),15-二烯（PD）的产生。我们进一步报道了构巢曲霉 PD 合酶基因（pbcA）的特征。在构巢曲霉中，pbcA 的过表达导致 PD 的产生，而 pbcA 的缺失则消除了 PD 的产生。 Fusarium fujikuroi 的 ent-贝壳杉烯合酶（cps/ks）和 Citrus unshiu 的γ-松油烯合酶的过表达分别导致了-ent-贝壳杉烯和γ-松油烯的产生。构巢曲霉是一种真菌模式生物，与其他工业相关的 Aspergillus 物种密切相关。构巢曲霉是许多次生代谢物的已知生产者，但它生产异源单萜和二萜化合物的能力尚未得到表征。在这里，我们表明构巢曲霉能够异源生产萜类化合物，因此具有作为工业相关化合物生产宿主的潜力。这里报道的遗传工程原理也可应用于其他 Aspergillus 属。
