Genome Assembly and Genetic Traits of the Pleuromutilin-Producer Clitopilus passeckerianus DSM1602

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Thomas Schafhauser - , University of Tübingen (Author)
  • Daniel Wibberg - , Bielefeld University, Jülich Research Centre (Author)
  • Antonia Binder - , Chair of General Microbiology, University of Tübingen (Author)
  • Christian Rückert - , Bielefeld University (Author)
  • Tobias Busche - , Bielefeld University (Author)
  • Wolfgang Wohlleben - , University of Tübingen, German Center for Infection Research (Author)
  • Jörn Kalinowski - , Bielefeld University (Author)

Abstract

The gilled mushroom Clitopilus passeckerianus (Entolomataceae, Agaricales, Basidiomycota) is well known to produce the terpenoid pleuromutilin, which is the biotechnological basis for medically important antibiotics such as lefamulin and retapamulin. Their unique mode of action and good tolerance entails an increasing demand of pleuromutilin-derived antibiotics in veterinary and human health care. Surprisingly, despite their pharmaceutical importance, no genome sequence is available of any pleuromutilin-producing fungus. Here, we present the high-quality draft genome sequence of the pleuromutilin-producer C. passeckerianus DSM1602 including functional genome annotation. More precisely, we employed a hybrid assembly strategy combining Illumina sequencing and Nanopore sequencing to assemble the mitochondrial genome as well as the nuclear genome. In accordance with the dikaryotic state of the fungus, the nuclear genome has a diploid character. Interestingly, the mitochondrial genome appears duplicated. Bioinformatic analysis revealed a versatile secondary metabolism with an emphasis on terpenoid biosynthetic enzymes in C. passeckerianus and also in related strains. Two alleles of biosynthetic gene clusters for pleuromutilin were found in the genome of C. passeckerianus. The pleuromutilin genes were reassembled with yeast-specific elements for heterologous expression in Saccharomyces cerevisiae. Our work lays the foundation for metabolic strain engineering towards higher yields of the valuable compound pleuromutilin.

Details

Original languageEnglish
Article number862
JournalJournal of Fungi
Volume8
Issue number8
Publication statusPublished - Aug 2022
Peer-reviewedYes

Keywords

Sustainable Development Goals

Keywords

  • Agaricales, biosynthetic gene cluster, Clitopilus, genome assembly, heteroplasmic mycelium, Illumina sequencing, mitochondrial genome, Nanopore sequencing, pleuromutilin, terpene synthase