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Characterization of an evolved carotenoids hyper-producer of Saccharomyces cerevisiae through bioreactor parameter optimization and Raman spectroscopy

  • Fermentation, Cell Culture and Bioengineering
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

An evolutionary engineering approach for enhancing heterologous carotenoids production in an engineered Saccharomyces cerevisiae strain was used previously to isolate several carotenoids hyper-producers from the evolved populations. β-Carotene production was characterized in the parental and one of the evolved carotenoids hyper-producers (SM14) using bench-top bioreactors to assess the impact of pH, aeration, and media composition on β-carotene production levels. The results show that with maintaining a low pH and increasing the carbon-to-nitrogen ratio (C:N) from 8.8 to 50 in standard YNB medium, a higher β-carotene production level at 25.52 ± 2.15 mg β-carotene g−1 (dry cell weight) in the carotenoids hyper-producer was obtained. The increase in C:N ratio also significantly increased carotenoids production in the parental strain by 298 % [from 5.68 ± 1.24 to 22.58 ± 0.11 mg β-carotene g−1 (dcw)]. In this study, it was shown that Raman spectroscopy is capable of monitoring β-carotene production in these cultures. Raman spectroscopy is adaptable to large-scale fermentations and can give results in near real-time. Furthermore, we found that Raman spectroscopy was also able to measure the relative lipid compositions and protein content of the parental and SM14 strains at two different C:N ratios in the bioreactor. The Raman analysis showed a higher total fatty acid content in the SM14 compared with the parental strain and that an increased C:N ratio resulted in significant increase in total fatty acid content of both strains. The data suggest a positive correlation between the yield of β-carotene per biomass and total fatty acid content of the cell.

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Acknowledgments

This research was funded in part by National Science Foundation NSF MCB-1054276. The authors wish to thank Dr. Alim Dewan and Professor M. Nazmul Karim for their assistance with the bioreactor.

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Authors and Affiliations

  1. Department of Chemical Engineering, Texas A&M University, College Station, TX, USA

    Michelle L. Olson, James Johnson, Luis H. Reyes & Katy C. Kao

  2. Department of Biological Systems Engineering, Virginia Tech, Blacksburg, VA, USA

    William F. Carswell & Ryan S. Senger

  3. Institute for the Study of Inborn Errors of Metabolism, School of Sciences, Pontificia Universidad Javeriana, Bogotá D.C., Colombia

    Luis H. Reyes

Authors
  1. Michelle L. Olson
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  2. James Johnson
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  3. William F. Carswell
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  4. Luis H. Reyes
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  5. Ryan S. Senger
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  6. Katy C. Kao
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Correspondence to Katy C. Kao.

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Olson, M.L., Johnson, J., Carswell, W.F. et al. Characterization of an evolved carotenoids hyper-producer of Saccharomyces cerevisiae through bioreactor parameter optimization and Raman spectroscopy. J Ind Microbiol Biotechnol 43, 1355–1363 (2016). https://doi.org/10.1007/s10295-016-1808-9

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  • DOI: https://doi.org/10.1007/s10295-016-1808-9

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