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Enhanced FAD Production in Eremothecium ashbyi with Statistically Optimized Oxidative Stress Modulators

Received: 12 December 2019    Accepted: 27 December 2019    Published: 31 January 2020
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Abstract

Riboflavin (vitamin B2) and flavin adenine dinucleotide (FAD) are valuable biological flavins mainly used as animal feed supplement and in pharmaceutical formulation. Eremothecium ashbyi is a natural overproducer of FAD along with riboflavin. Flavins overproduction was found to be strongly correlated with stress induction but stress response in E.ashbyi is completely unexplored. Hence, in the present study, flavins production in presence of four oxidative stress modulators (menadione, vitamin E, cAMP, H2O2) was investigated. Their levels were statistically optimized to maximize the flavins production. The regression model obtained from Central Composite Design indicated that maximum flavins production occurred when 1.09 µM menadione and 1.12 µM vitamin E were supplemented to 24-hour grown fungus. Upon supplementation, total flavins production was significantly increased by 1.34-fold and FAD production was increased by 2.18-fold over untreated control. Intracellular – reactive oxygen species (ROS) level was increased and reduced glutathione (GSH) and oxidized glutathione (GSSG) ratio was decreased indicating oxidative burst experienced by the cell. Furthermore, a concomitant increase in specific activity of glutathione reductase and FAD synthetase was observed. For the first time this study showed that among flavins, FAD was majorly increased along with FAD synthetase activity as a response to oxidative stress. These results indicated that the oxidative stress and flavins overproduction was centered on FAD regulation.

Published in International Journal of Microbiology and Biotechnology (Volume 5, Issue 1)
DOI 10.11648/j.ijmb.20200501.12
Page(s) 7-15
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

Eremothecium ashbyi, Riboflavin, FAD, Central Composite Design (CCD), Glutathione, Oxidative Stress

References
[1] Colibus L. and Mattevi A. (2006) New frontiers in structural flavoenzymology, Curr. Opin. Struct. Biol. 16: 722–728.
[2] Yatsyshyn V., Fedorovych D. and Sibirny A. (2014) Metabolic and bioprocess engineering of the yeast Candida famata for FAD production, J. Ind. Microbiol. Biotechnol. 41: 823–835.
[3] Gonzalez-Cabo P., Ros S. and Palau F. (2014) Flavin adenine dinucleotide rescues the phenotype of frataxin deficiency, PLoS One 5: 1–10.
[4] Huang Y., Liu S., Yen C., et. al. (2009) Thapsigargin and flavin adenine dinucleotide ex vivo treatment rescues trafficking-defective gp91phox in chronic granulomatous disease leukocytes, Free Radic. Biol. Med. 47: 932–940.
[5] Ashby S. and Nowell W. (1926) The Fungi of Stigmatomycosis, Ann. Bot. 40: 69–84.
[6] Kato T. and Park E. (2012) Riboflavin production by Ashbya gossypii, Biotechnol. Lett. 34: 611–618.
[7] Abbas C. and Sibirny A. (2011) Genetic Control of Biosynthesis and Transport of Riboflavin and Flavin Nucleotides and Construction of Robust Biotechnological Producers, Microbiol. Mol. Biol. Rev. 75: 321–360.
[8] Ozbas T. and Kutsal T. (1986) Comparative study of riboflavin production from two microorganisms: Eremothecium ashbyi and Ashbya gossypii, Enzyme Microb. Technol. 8: 593–596.
[9] Protchenko O., Boretsky B., Romanyuk T., et. al. (2000) Oversynthesis of riboflavin by yeast Pichia guilliermondii in response to oxidative stress, Ukr. Biokhimichnyi Zhurnal 72: 19–23.
[10] Schlösser T., Wiesenburg A., Gätgens C., et. al. (2007) Growth stress triggers riboflavin overproduction in Ashbya gossypii, Appl. Microbiol. Biotechnol. 76: 569–578.
[11] Kavitha S. and Chandra T. S. (2009) Effect of vitamin E and menadione supplementation on riboflavin production and stress parameters in Ashbya gossypii, Process Biochem. 44: 934–938.
[12] Stahmann K., Arst H., Althofer H. et. al. (2001) Riboflavin, overproduced during sporulation of Ashbya gossypii, protects its hyaline spores against ultraviolet light, Environ. Microbiol. 3: 545–550.
[13] Kavitha S. and Chandra T. S. (2014) Oxidative Stress Protection and Glutathione Metabolism in Response to Hydrogen Peroxide and Menadione in Riboflavinogenic Fungus Ashbya gossypii, Appl. Biochem. Biotechnol. 174: 2307–2325.
[14] Walther A. and Wendland J. (2012) Yap1-dependent oxidative stress response provides a link to riboflavin production in Ashbya gossypii, Fungal Genet. Biol. 49: 697–707.
[15] Ashoori M. and Saedisomeolia A. (2014) Riboflavin (vitamin B2) and oxidative stress: a review, Br. J. Nutr. 111: 1985–1991.
[16] Patel M. and Chnadra T. S. (2019) Metabolic engineering of Ashbya gossypii for enhanced FAD production through promoter replacement of FMN1 gene, Enzyme Microb. Technol. 133.
[17] Ayar-Kayali H., Ozer N. and Tarhan L. (2002) Intracellular superoxide dismutase, catalase, and glutathione peroxidase activities and membrane lipid peroxide levels in Fusarium acuminatum upon environmental changes in a defined medium, Arch. Biochem. Biophys. 400: 265–272.
[18] Tietze F. (1969) Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: Applications to mammalian blood and other tissues, Anal. Biochem. 27: 502–522.
[19] Mavis R. and Stellwagen E. (1968) Purification and Subunit of Glutathione Reductase from Bakers Yeast, J. Biol. Chem. 243: 809–814.
[20] Rastogi R., Singh S., Hader D., et. al. (2010) Detection of reactive oxygen species (ROS) by the oxidant-sensing probe 2’,7’-dichlorodihydrofluorescein diacetate in the cyanobacterium Anabaena variabilis PCC 7937, Biochem. Biophys. Res. Commun. 397: 603–607.
[21] Orth A., Sfarra A., Pell E., et. al. (1993) Assessing the Involvement of Free Radicals in Fungicide Toxicity Using α-Tocopherol Analogs, Pestic. Biochem. Physiol. 47: 134–141.
[22] Wong-Ekkabut J., Xu Z., Triampo W., et. al. (2007) Effect of lipid peroxidation on the properties of lipid bilayers: a molecular dynamics study, Biophys. J. 93: 4225–36.
[23] Grant C. (2001) Role of the glutathione/glutaredoxin and thioredoxin systems in yeast growth and response to stress conditions, Mol. Microbiol. 39: 533–541.
[24] Tsukihara K., Minoura K. and Izumiya M. (1960) Studies on the industrial production of flavin-adenine dinucleotide. Preparation of mycelium of Eremothecium ashbyi as a raw material containing flavin-adenine dinucleotide, J. Vitaminol. (Kyoto). 6: 68–76.
[25] Lim S., Choi J. and Park E. (2001) Microbial Production of Riboflavin Using Riboflavin Overproducers Ashbya gossypii, Bacillus subtilis, and Candida famata: An Overview, Biotechnol. Bioprocess Eng. 6: 75–88.
[26] Grant C., Collinson L., Roe J. et. al. (1996) Yeast glutathione reductase is required for protection against oxidative stress and is a target gene for yAP-1 transcriptional regulation, Mol. Microbiol. 21: 171–179.
[27] Wojtaszek P. (1997) Oxidative burst: an early plant response to pathogen infection, Biochem. J. 322: 681–92.
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  • APA Style

    Manan Vipulbhai Patel, Chandra Sainathan. (2020). Enhanced FAD Production in Eremothecium ashbyi with Statistically Optimized Oxidative Stress Modulators. International Journal of Microbiology and Biotechnology, 5(1), 7-15. https://doi.org/10.11648/j.ijmb.20200501.12

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    ACS Style

    Manan Vipulbhai Patel; Chandra Sainathan. Enhanced FAD Production in Eremothecium ashbyi with Statistically Optimized Oxidative Stress Modulators. Int. J. Microbiol. Biotechnol. 2020, 5(1), 7-15. doi: 10.11648/j.ijmb.20200501.12

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    AMA Style

    Manan Vipulbhai Patel, Chandra Sainathan. Enhanced FAD Production in Eremothecium ashbyi with Statistically Optimized Oxidative Stress Modulators. Int J Microbiol Biotechnol. 2020;5(1):7-15. doi: 10.11648/j.ijmb.20200501.12

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  • @article{10.11648/j.ijmb.20200501.12,
      author = {Manan Vipulbhai Patel and Chandra Sainathan},
      title = {Enhanced FAD Production in Eremothecium ashbyi with Statistically Optimized Oxidative Stress Modulators},
      journal = {International Journal of Microbiology and Biotechnology},
      volume = {5},
      number = {1},
      pages = {7-15},
      doi = {10.11648/j.ijmb.20200501.12},
      url = {https://doi.org/10.11648/j.ijmb.20200501.12},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmb.20200501.12},
      abstract = {Riboflavin (vitamin B2) and flavin adenine dinucleotide (FAD) are valuable biological flavins mainly used as animal feed supplement and in pharmaceutical formulation. Eremothecium ashbyi is a natural overproducer of FAD along with riboflavin. Flavins overproduction was found to be strongly correlated with stress induction but stress response in E.ashbyi is completely unexplored. Hence, in the present study, flavins production in presence of four oxidative stress modulators (menadione, vitamin E, cAMP, H2O2) was investigated. Their levels were statistically optimized to maximize the flavins production. The regression model obtained from Central Composite Design indicated that maximum flavins production occurred when 1.09 µM menadione and 1.12 µM vitamin E were supplemented to 24-hour grown fungus. Upon supplementation, total flavins production was significantly increased by 1.34-fold and FAD production was increased by 2.18-fold over untreated control. Intracellular – reactive oxygen species (ROS) level was increased and reduced glutathione (GSH) and oxidized glutathione (GSSG) ratio was decreased indicating oxidative burst experienced by the cell. Furthermore, a concomitant increase in specific activity of glutathione reductase and FAD synthetase was observed. For the first time this study showed that among flavins, FAD was majorly increased along with FAD synthetase activity as a response to oxidative stress. These results indicated that the oxidative stress and flavins overproduction was centered on FAD regulation.},
     year = {2020}
    }
    

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  • TY  - JOUR
    T1  - Enhanced FAD Production in Eremothecium ashbyi with Statistically Optimized Oxidative Stress Modulators
    AU  - Manan Vipulbhai Patel
    AU  - Chandra Sainathan
    Y1  - 2020/01/31
    PY  - 2020
    N1  - https://doi.org/10.11648/j.ijmb.20200501.12
    DO  - 10.11648/j.ijmb.20200501.12
    T2  - International Journal of Microbiology and Biotechnology
    JF  - International Journal of Microbiology and Biotechnology
    JO  - International Journal of Microbiology and Biotechnology
    SP  - 7
    EP  - 15
    PB  - Science Publishing Group
    SN  - 2578-9686
    UR  - https://doi.org/10.11648/j.ijmb.20200501.12
    AB  - Riboflavin (vitamin B2) and flavin adenine dinucleotide (FAD) are valuable biological flavins mainly used as animal feed supplement and in pharmaceutical formulation. Eremothecium ashbyi is a natural overproducer of FAD along with riboflavin. Flavins overproduction was found to be strongly correlated with stress induction but stress response in E.ashbyi is completely unexplored. Hence, in the present study, flavins production in presence of four oxidative stress modulators (menadione, vitamin E, cAMP, H2O2) was investigated. Their levels were statistically optimized to maximize the flavins production. The regression model obtained from Central Composite Design indicated that maximum flavins production occurred when 1.09 µM menadione and 1.12 µM vitamin E were supplemented to 24-hour grown fungus. Upon supplementation, total flavins production was significantly increased by 1.34-fold and FAD production was increased by 2.18-fold over untreated control. Intracellular – reactive oxygen species (ROS) level was increased and reduced glutathione (GSH) and oxidized glutathione (GSSG) ratio was decreased indicating oxidative burst experienced by the cell. Furthermore, a concomitant increase in specific activity of glutathione reductase and FAD synthetase was observed. For the first time this study showed that among flavins, FAD was majorly increased along with FAD synthetase activity as a response to oxidative stress. These results indicated that the oxidative stress and flavins overproduction was centered on FAD regulation.
    VL  - 5
    IS  - 1
    ER  - 

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Author Information
  • Department of Biotechnology, Indian Institute of Technology Madras, Chennai, India

  • Department of Biotechnology, Indian Institute of Technology Madras, Chennai, India

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