Ascomycete Aspergillus oryzae is an efficient expression host for production of basidiomycete terpenes by using genomic DNA sequences

Shota Nagamine, Chengwei Liu, Jumpei Nishishita, Takuto Kozaki, Kaho Sogahata, Yoshiro Sato, Atsushi Minami, Taro Ozaki, Claudia Schmidt-Dannert, Jun ichi Maruyama, Hideaki Oikawa

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

Basidiomycete fungi are an attractive resource for biologically active natural products for use in pharmaceutically relevant compounds. Recently, genome projects on mushroom fungi have provided a great deal of biosynthetic gene cluster information. However, functional analyses of the gene clusters for natural products were largely unexplored because of the difficulty of cDNA preparation and lack of gene manipulation tools for basidiomycete fungi. To develop a versatile host for basidiomycete genes, we examined gene expression using genomic DNA sequences in the robust ascomycete host Aspergillus oryzae, which is frequently used for the production of metabolites from filamentous fungi. Exhaustive expression of 30 terpene synthase genes from the basidiomycetes Clitopilus pseudo-pinsitus and Stereum hirsutum showed two splicing patterns, i.e., completely spliced cDNAs giving terpenes (15 cases) and mostly spliced cDNAs, indicating that A. oryzae correctly spliced most introns at the predicted positions and lengths. The mostly spliced cDNAs were expressed after PCR-based removal of introns, resulting in the successful production of terpenes (14 cases). During this study, we observed relatively frequent mispredictions in the automated program. Hence, the complementary use of A. oryzae expression and automated prediction will be a powerful tool for genome mining.

Original languageEnglish (US)
Article numbere00409-19
JournalApplied and environmental microbiology
Volume85
Issue number15
DOIs
StatePublished - 2019

Bibliographical note

Funding Information:
This work was financially supported by Japan Society for the Promotion of Science grants-in-aid JP15H01835 to H.O., JP16H03277 and JP16H06446 to A.M., and JP18K14342 to C.L.

Publisher Copyright:
© 2019 American Society for Microbiology.

Keywords

  • Aspergillus oryzae
  • Basidiomycete fungi
  • Heterologous expression
  • Terpene synthase
  • Terpenes

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