Antibacterial diphenyl ether production induced by co-culture of Aspergillus nidulans and Aspergillus fumigatus

Appl Microbiol Biotechnol. 2022 Jun;106(11):4169-4185. doi: 10.1007/s00253-022-11964-5. Epub 2022 May 21.


Fungi are a rich source of secondary metabolites with potent biological activities. Co-culturing a fungus with another microorganism has drawn much attention as a practical method for stimulating fungal secondary metabolism. However, in most cases, the molecular mechanisms underlying the activation of secondary metabolite production in co-culture are poorly understood. To elucidate such a mechanism, in this study, we established a model fungal-fungal co-culture system, composed of Aspergillus nidulans and Aspergillus fumigatus. In the co-culture of A. nidulans and A. fumigatus, production of antibacterial diphenyl ethers was enhanced. Transcriptome analysis by RNA-sequencing showed that the co-culture activated expression of siderophore biosynthesis genes in A. fumigatus and two polyketide biosynthetic gene clusters (the ors and cic clusters) in A. nidulans. Gene disruption experiments revealed that the ors cluster is responsible for diphenyl ether production in the co-culture. Interestingly, the ors cluster was previously reported to be upregulated by co-culture of A. nidulans with the bacterium Streptomyces rapamycinicus; orsellinic acid was the main product of the cluster in that co-culture. In other words, the main product of the ors cluster was different in fungal-fungal and bacterial-fungal co-culture. The genes responsible for biosynthesis of the bacterial- and fungal-induced polyketides were deduced using a heterologous expression system in Aspergillus oryzae. The molecular genetic mechanisms that trigger biosynthesis of two different types of compounds in A. nidulans in response to the fungus and the bacterium were demonstrated, which provides an insight into complex secondary metabolic response of fungi to microorganisms. KEY POINTS: • Co-culture of two fungal species triggered antibiotic diphenyl ether production. • The co-culture affected expression levels of several genes for secondary metabolism. • Gene cluster essential for induction of the antibiotics production was determined.

Keywords: Aspergillus fumigatus; Aspergillus nidulans; Biosynthesis; Co-culture; Diphenyl ether; Secondary metabolism.

MeSH terms

  • Anti-Bacterial Agents / metabolism
  • Aspergillus fumigatus / genetics
  • Aspergillus nidulans* / genetics
  • Aspergillus nidulans* / metabolism
  • Coculture Techniques
  • Gene Expression Regulation, Fungal
  • Multigene Family
  • Phenyl Ethers / metabolism
  • Polyketides* / metabolism


  • Anti-Bacterial Agents
  • Phenyl Ethers
  • Polyketides
  • phenyl ether