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Improved α-amylase production by Aspergillus oryzae after a double deletion of genes involved in carbon catabolite repression

  • Applied genetics and molecular biotechnology
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Abstract

In filamentous fungi, the expression of secretory glycoside hydrolase encoding genes, such as those for amylases, cellulases, and xylanases, is generally repressed in the presence of glucose. CreA and CreB have been observed to be regulating factors for carbon catabolite repression. In this study, we generated single and double deletion creA and/or creB mutants in Aspergillus oryzae. The α-amylase activities of each strain were compared under various culture conditions. For the wild-type strain, mRNA levels of α-amylase were markedly decreased in the later stage of submerged culture under inducing conditions, whereas this reduced expression was not observed for single creA and double creA/creB deletion mutants. In addition, α-amylase activity of the wild-type strain was reduced in submerged culture containing high concentrations of inducing sugars, whereas all constructed mutants showed higher α-amylase activities. In particular, the α-amylase activity of the double deletion mutant in a medium containing 5 % starch was >10-fold higher than that of the wild-type strain under the same culture conditions. In solid-state cultures using wheat bran as a substrate, the α-amylase activities of single creA and double deletion mutants were >2-fold higher than that of the wild-type strain. These results suggested that deleting both creA and creB resulted in dramatic improvements in the production of secretory glycoside hydrolases in filamentous fungi.

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Acknowledgments

We thank Osamu Mizutani for kindly providing the ∆ligD::loxP pyrG − mutant strain. We thank Youhei Kudo and Tetsuya Hiramoto for ∆creA/pYES2 plasmid construction. This study was supported by the Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN).

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Authors and Affiliations

  1. Laboratory of Bioindustrial Genomics, Department of Bioindustrial Informatics and Genomics, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai, 981-8555, Japan

    Sakurako Ichinose, Mizuki Tanaka, Takahiro Shintani & Katsuya Gomi

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  1. Sakurako Ichinose
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  2. Mizuki Tanaka
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  3. Takahiro Shintani
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  4. Katsuya Gomi
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Correspondence to Katsuya Gomi.

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Sakurako Ichinose and Mizuki Tanaka contributed equally to this article.

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Ichinose, S., Tanaka, M., Shintani, T. et al. Improved α-amylase production by Aspergillus oryzae after a double deletion of genes involved in carbon catabolite repression. Appl Microbiol Biotechnol 98, 335–343 (2014). https://doi.org/10.1007/s00253-013-5353-4

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  • DOI: https://doi.org/10.1007/s00253-013-5353-4

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