Identification of a polyketide biosynthesis gene cluster by transcriptional regulator activation in Aspergillus terreus

Shen Tang; Ping Men; Wei Zhang; Hui Li; Zhimin Li; Xuenian Huang; Xuefeng Lu

doi:10.1016/j.fgb.2022.103690

Identification of a polyketide biosynthesis gene cluster by transcriptional regulator activation in Aspergillus terreus

Fungal Genet Biol. 2022 May:160:103690. doi: 10.1016/j.fgb.2022.103690. Epub 2022 Mar 26.

Authors

Shen Tang¹, Ping Men², Wei Zhang², Hui Li³, Zhimin Li⁴, Xuenian Huang⁵, Xuefeng Lu⁶

Affiliations

¹ 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.
² 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.
³ 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.
⁴ College of Bioscience and Bioengineering, Jiangxi Engineering Laboratory for the Development and Utilization of Agricultural Microbial Resources, Jiangxi Agricultural University, Nanchang 330045, China.
⁵ 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. Electronic address: huangxn@qibebt.ac.cn.
⁶ Shandong Provincial Key Laboratory of Synthetic Biology, China; Key Laboratory of Biofuels, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No. 189 Songling Road, Qingdao 266101, 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; Marine biology and Biotechnology Laboratory, Qingdao National Laboratory for Marine Science and Technology, Wenhai Rd 1, Aoshanwei, Qingdao, China. Electronic address: lvxf@qibebt.ac.cn.

PMID: 35351612
DOI: 10.1016/j.fgb.2022.103690

Abstract

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.

Keywords: Aspergillus terreus; Biosynthetic pathway; Naphthoquinone; Polyketide synthase; Silent gene cluster activation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aspergillus
Multigene Family
Naphthoquinones*
Polyketide Synthases / genetics
Polyketide Synthases / metabolism
Polyketides* / metabolism
Secondary Metabolism / genetics

Substances

Naphthoquinones
Polyketides
Polyketide Synthases

Supplementary concepts

Aspergillus terreus