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Applied Microbiology and Biotechnology

, Volume 100, Issue 13, pp 5859–5868 | Cite as

The C2H2-type transcription factor, FlbC, is involved in the transcriptional regulation of Aspergillus oryzae glucoamylase and protease genes specifically expressed in solid-state culture

  • Mizuki Tanaka
  • Midori Yoshimura
  • Masahiro Ogawa
  • Yasuji Koyama
  • Takahiro Shintani
  • Katsuya GomiEmail author
Applied genetics and molecular biotechnology

Abstract

Aspergillus oryzae produces a large amount of secreted proteins in solid-state culture, and some proteins such as glucoamylase (GlaB) and acid protease (PepA) are specifically produced in solid-state culture, but rarely in submerged culture. From the disruption mutant library of A. oryzae transcriptional regulators, we successfully identified a disruption mutant showing an extremely low production level of GlaB but a normal level of α-amylase production. This strain was a disruption mutant of the C2H2-type transcription factor, FlbC, which is reported to be involved in the regulation of conidiospore development. Disruption mutants of other upstream regulators comprising a conidiation regulatory network had no apparent effect on GlaB production in solid-state culture. In addition to GlaB, the production of acid protease in solid-state culture was also markedly decreased by flbC disruption. Northern blot analyses revealed that transcripts of glaB and pepA were significantly decreased in the flbC disruption strain. These results suggested that FlbC is involved in the transcriptional regulation of genes specifically expressed under solid-state cultivation conditions, possibly independent of the conidiation regulatory network.

Keywords

Aspergillus oryzae Solid-state culture Transcription factor FlbC Glucoamylase Acid protease Gene expression regulation Conidiospore development 

Notes

Acknowledgments

We are grateful to Yoji Hata for kindly providing the anti-GlaB antibody and Yukihiro Nakamura for technical advice about the tyrosinase activity assay. We also thank Takeshi Akao for technical advice about total RNA extraction from A. oryzae cultured in solid-state culture and Osamu Mizutani for kindly providing the ∆ligD::loxP pyrG strain. This study was supported by the Program for Promotion of Basic and Applied Researches for Innovations in Bio-oriented Industry and the Science and Technology Research Promotion Program for Agriculture, Forestry, Fisheries and Food Industry.

Compliance with ethical standards

Ethical approval

This article does not contain any study with human participants or animals performed by any of the authors.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

253_2016_7419_MOESM1_ESM.pdf (372 kb)
ESM 1 (PDF 371 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  1. 1.Laboratory of Bioindustrial Genomics, Department of Bioindustrial Informatics and Genomics, Graduate School of Agricultural ScienceTohoku UniversitySendaiJapan
  2. 2.Noda Institute for Scientific ResearchNodaJapan

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