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Induction of Oxidative Stress in Sirtuin Gene-Disrupted Ashbya gossypii Mutants Overproducing Riboflavin

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Abstract

AgHST1 and AgHST3 genes encode sirtuins that are NAD+-dependent protein deacetylases. According to previous reports, their disruption leads to the overproduction of riboflavin in Ashbya gossypii. In this study, we investigated the potential causes of riboflavin overproduction in the AgHST1Δ and AgHST3Δ mutant strains of A. gossypii. The generation of reactive oxygen species was increasd in the mutants compared to in WT. Additionally, membrane potential was lower in the mutants than in WT. The NAD+/NADH ratio in AgHST1Δ mutant strain was lower than that in WT; however, the NAD+/NADH ratio in AgHST3Δ was slightly higher than that in WT. AgHST1Δ mutant strain was more sensitive to high temperatures and hydroxyurea treatment than WT or AgHST3Δ. Expression of the AgGLR1 gene, encoding glutathione reductase, was substantially decreased in AgHST1Δ and AgHST3Δ mutant strains. The addition of N-acetyl-L-cysteine, an antioxidant, suppressed the riboflavin production in the mutants, indicating that it was induced by oxidative stress. Therefore, high oxidative stress resulting from the disruption of sirtuin genes induces riboflavin overproduction in AgHST1Δ and AgHST3Δ mutant strains. This study established that oxidative stress is an important trigger for riboflavin overproduction in sirtuin gene-disrupted mutant strains of A. gossypii and helped to elucidate the mechanism of riboflavin production in A. gossypii.

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All data and materials analyzed in this study are shown in this published article.

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Acknowledgements

We would like to thank Kohei Kurabayashi (Department of Applied Life Science, Faculty of Agriculture, Shizuoka University) to help our response to reviewers with additional experiments.

Funding

This study was supported by JSPS KAKENHI (Grant-in-Aid for Scientific Research (C); Grant Number JP21K05390).

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Authors and Affiliations

  1. Molecular and Biological Function Research Core, Research Institute of Green Science and Technology, Shizuoka University, Ohya 836, Suruga-ku, Shizuoka, Japan

    Tatsuya Kato & Enoch Y. Park

  2. Department of Agriculture, Graduate School of Integrated Science and Technology, Shizuoka University, Ohya 836, Suruga-ku, Shizuoka, Japan

    Tatsuya Kato, Junya Azegami, Mai Kano & Enoch Y. Park

  3. Department of Applied Life Science, Faculty of Agriculture, Shizuoka University, Ohya 836, Suruga-ku, Shizuoka, Japan

    Tatsuya Kato & Enoch Y. Park

  4. Institute of Bioproduct Development (IBD), Universiti Teknologi Malaysia (UTM), 81310 UTM, Johor Bahru, Malaysia

    Hesham A. El Enshasy

  5. City of Scientific Research and Technology Applications, New Borg Al Arab, Alexandria, Egypt

    Hesham A. El Enshasy

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  1. Tatsuya Kato
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  2. Junya Azegami
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Contributions

Study conception and design were performed by TK, HAEE, and EYP. Material preparation, data collection, and analysis were performed by JA, MK, HAEE, and EYP. The first draft of the manuscript was written by TK, HAEE and EYP. All authors read and approved the final manuscript.

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Correspondence to Tatsuya Kato.

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Kato, T., Azegami, J., Kano, M. et al. Induction of Oxidative Stress in Sirtuin Gene-Disrupted Ashbya gossypii Mutants Overproducing Riboflavin. Mol Biotechnol 66, 1144–1153 (2024). https://doi.org/10.1007/s12033-023-01012-6

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