Improved α-amylase production by dephosphorylation mutation of CreD, an arrestin-like protein required for glucose-induced endocytosis of maltose permease and carbon catabolite derepression in Aspergillus oryzae

Mizuki Tanaka, Tetsuya Hiramoto, Hinako Tada, Takahiro Shintani, Katsuya Gomi

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Aspergillus oryzae produces copious amount of amylolytic enzymes, and MalP, a major maltose permease, is required for the expression of amylase-encoding genes. The expression of these genes is strongly repressed by carbon catabolite repression (CCR) in the presence of glucose. MalP is transported from the plasma membrane to the vacuole by endocytosis, which requires the homolog of E6-AP carboxyl terminus ubiquitin ligase HulA, an ortholog of yeast Rsp5. In yeast, arrestin-like proteins mediate endocytosis as adaptors of Rsp5 and transporters. In the present study, we examined the involvement of CreD, an arrestin-like protein, in glucoseinduced MalP endocytosis and CCR of amylase-encoding genes. Deletion of creD inhibited the glucose-induced endocytosis of MalP, and CreD showed physical interaction with HulA. Phosphorylation of CreD was detected by Western blotting, and two serine residues were determined as the putative phosphorylation sites. However, the phosphorylation state of the serine residues did not regulate MalP endocytosis and its interaction with HulA. Although α-amylase production was significantly repressed by creD deletion, both phosphorylation and dephosphorylation mimics of CreD had a negligible effect on α-amylase activity. Interestingly, dephosphorylation of CreD was required for CCR relief of amylase genes that was triggered by disruption of the deubiquitinating enzyme-encoding gene creB. The α-amylase activity of the creB mutant was 1.6-fold higher than that of the wild type, and the dephosphorylation mimic of CreD further improved the α-amylase activity by 2.6-fold. These results indicate that a combination of the dephosphorylation mutation of CreD and creB disruption increased the production of amylolytic enzymes in A. oryzae.

Original languageEnglish
Article numbere00592-17
JournalApplied and environmental microbiology
Volume83
Issue number13
DOIs
Publication statusPublished - 2017

Keywords

  • Arrestin-like protein
  • Aspergillus oryzae
  • Carbon catabolite repression
  • Dephosphorylation
  • Endocytosis
  • Ubiquitin ligase
  • α-amylase production

ASJC Scopus subject areas

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

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