Engineering Aspergillus nidulans for heterologous ent-kaurene and gamma-terpinene production
Engineering Aspergillus nidulans for heterologous ent-kaurene and gamma-terpinene production
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DOI：
10.1007/s00253-016-7517-5
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发表时间：
2016-07-01
影响因子：
5
通讯作者：
Nakari-Setala, Tiina
中科院分区：
文献类型：
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作者：
Bromann, Kirsi;Toivari, Mervi;Nakari-Setala, Tiina
萜烯是一种种类繁多的天然产物，具有广泛的生物活性和应用。由于这些化合物结构复杂，工业相关萜烯的化学生产可能既昂贵又耗时。在这里，我们研究了灰曲霉作为异源宿主生产单萜烯和二萜烯。之前，我们在a . nidulans中发现了一个新的二萜基因簇，并发现簇特异性转录因子（pbcR）的过表达导致了ent-pimara-8(14)，15-二烯（PD）的产生。我们报道了A. nidulans PD合成酶基因（pbcA）的进一步表征。在A. nidulans中，pbcA的过表达导致PD的产生，而pbcA的缺失则使PD的产生消失。Fusarium fujikuroi -kaurene合成酶（cps/ks）和Citrus unshiu -terpinene合成酶的过表达分别导致了-kaurene和-terpinene的产生。a . nidulans是一种真菌模式生物，是其他工业相关曲霉种的近亲。白莲属植物是许多次生代谢产物的已知生产者，但其产生异源单萜和二萜化合物的能力尚未被表征。在这里，我们证明了a . nidulans能够产生异源萜烯，因此具有作为工业相关化合物的生产宿主的潜力。本文报道的基因工程原理也可以应用于其他曲霉菌。
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.