通过转录调节因子激活在土曲霉中鉴定聚酮生物合成基因簇
Identification of a polyketide biosynthesis gene cluster by transcriptional regulator activation in Aspergillus terreus.
作者信息
Tang Shen, Men Ping, Zhang Wei, Li Hui, Li Zhimin, Huang Xuenian, Lu Xuefeng
机构信息
Shandong Provincial Key Laboratory of Synthetic Biology, China; Shandong Energy Institute, No. 189 Songling Road, Qingdao 266101, China; Qingdao New Energy Shandong Laboratory, No. 189 Songling Road, Qingdao 266101, China; College of Bioscience and Bioengineering, Jiangxi Engineering Laboratory for the Development and Utilization of Agricultural Microbial Resources, Jiangxi Agricultural University, Nanchang 330045, China.
Shandong Provincial Key Laboratory of Synthetic Biology, China; Shandong Energy Institute, No. 189 Songling Road, Qingdao 266101, China; Qingdao New Energy Shandong Laboratory, No. 189 Songling Road, Qingdao 266101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
出版信息
Fungal Genet Biol. 2022 May;160:103690. doi: 10.1016/j.fgb.2022.103690. Epub 2022 Mar 26.
In filamentous fungi, the secondary metabolism is environmentally sensitive. Most of the biosynthetic gene clusters of secondary metabolites are silent under laboratory conditions. In this study, a highly conserved naphthopyrone PKS ATEG_06206 was identified in the genome sequence of A. terreus MEFC01. This gene is silent under laboratory conditions. To study the function of this PKS, we activated the silent biosynthetic pathway by replacing the promoter of cluster-specific transcriptional factor ATEG_06205 in A. terreus MEFC01. With this strategy, we confirmed that the products of this cryptic PKS are naphthoquinones. These naphthoquinones are soluble pigments, which could be secreted into the agar medium and culture broth. For this reason, the colour of mycelium and conidia of the activated mutant was significantly darker than that of the parental strain. The gene cluster and biosynthetic pathway were further elucidated through the reverse genetics approach and enzymatic assay in vitro.
在丝状真菌中，次级代谢对环境敏感。大多数次级代谢产物的生物合成基因簇在实验室条件下是沉默的。在本研究中，在土曲霉MEFC01的基因组序列中鉴定出一个高度保守的萘并吡喃酮聚酮合酶ATEG_06206。该基因在实验室条件下是沉默的。为了研究该聚酮合酶的功能，我们通过替换土曲霉MEFC01中簇特异性转录因子ATEG_06205的启动子来激活沉默的生物合成途径。通过该策略，我们证实了这个隐秘聚酮合酶的产物是萘醌。这些萘醌是可溶性色素，可分泌到琼脂培养基和培养液中。因此，激活突变体的菌丝体和分生孢子的颜色明显比亲本菌株深。通过反向遗传学方法和体外酶促测定进一步阐明了基因簇和生物合成途径。
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