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Production of perillic acid from orange essential oil by Yarrowia lipolytica using a top-aerated bioreactor

  • Biotechnology and Industrial Microbiology - Research Paper
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Abstract

R-(+)-Perillic acid, a promising anticancer and immunomodulatory agent, is the major product from the biotransformation of R-(+)-limonene-rich orange essential oil by the yeast Yarrowia lipolytica. Due to the abundance and low cost of orange essential oil, which is a byproduct of the citrus industry, we attempted to improve the biotransformation process by optimizing yeast cell mass production. Then, the whole process was transposed and adapted to a 2-L instrumented bioreactor. Cell mass production was optimized in shaker flasks using a statistical experimental design. The optimized medium (g·L−1: 22.9 glucose, 7.7 peptone, 4.1 yeast extract and 1.0 malt extract) resulted in a 13.0 g·L−1 final cell concentration and 0.18 g cell·L−1·h−1 productivity. A further increase to 18.0 g·L−1 was achieved in a 2-L bioreactor upon fed-batch culture. High-purity limonene bioconversion was performed in the same bioreactor utilizing top aeration to diminish terpene volatilization; as a result, 839.6 mg·L−1 perillic acid accumulated after 48 h. Under the same conditions, industrial orange essential oil afforded 806.4 mg·L−1 perillic acid. The yeast growth medium optimization resulted in a twofold increase in biomass accumulation and a reduction in growth medium nitrogen sources, which lowered the catalytic biomass production cost. Compared with conventional bottom aeration, the bioreactor top aeration strategy resulted in higher bioconversion rates. The conditions developed for high-purity limonene bioconversion were successfully applied to low-cost orange essential oil, showing the robustness of Y. lipolytica yeast.

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Acknowledgements

The authors are grateful to Daniel Santos Pereira and Maria Alice Santos Cerullo for the HPLC analysis and to Lucas Tupi Caldas Pereira for the valuable assistance in the bioreactor experiments.

Funding

This work was supported by the National Council for Scientific and Technological Development (CNPq) and the Pharmaceutical Technology Institute of the Oswaldo Cruz Foundation under grant PROEP/FAR/CNPq 407841/2017–2; Carlos Chagas Filho Foundation for Research Support of the State of Rio de Janeiro (FAPERJ) under grants E-26/110.856/2013 and E-26/101.210/2013; CNPq under grant 307766/2014/4; and PIBIC/CNPq/FIOCRUZ 2014–2016.

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Authors and Affiliations

  1. Chemistry Institute, Federal University of Rio de Janeiro, Rio de Janeiro, RJ, Brazil

    Felipe M. Knopp & Elba P. S. Bon

  2. School of Chemistry, Federal University of Rio de Janeiro, Rio de Janeiro, RJ, Brazil

    Rodrigo da Rocha Olivieri de Barros

  3. Institute of Drug Technology, Oswaldo Cruz Foundation, Rio de Janeiro, RJ, Brazil

    Bruna S. Drummond, Antonio Carlos Siani & Maria Antonieta Ferrara

Authors
  1. Felipe M. Knopp
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  2. Rodrigo da Rocha Olivieri de Barros
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  3. Bruna S. Drummond
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  4. Antonio Carlos Siani
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  5. Maria Antonieta Ferrara
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  6. Elba P. S. Bon
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Contributions

F.M. Knopp contributed to the design of the studies, performed the experiments, analysed the experimental data, and contributed to writing the manuscript; R.R.O. Barros contributed to the design of the optimization studies and the critical revision of the manuscript; B.S. Drummond contributed to the performance of the experiments; A.C. Siani acquired funding for this work and contributed to the critical revision of the manuscript; M.A. Ferrara contributed to the supervision, conception and design of the studies and writing of the manuscript; E.P.S. Bon contributed to the supervision, provided lab space, acquired funding for this work and contributed to the critical revision of the manuscript.

All authors read and approved the final manuscript.

Corresponding author

Correspondence to Elba P. S. Bon.

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Knopp, F.M., da Rocha Olivieri de Barros, R., Drummond, B.S. et al. Production of perillic acid from orange essential oil by Yarrowia lipolytica using a top-aerated bioreactor. Braz J Microbiol 54, 2663–2670 (2023). https://doi.org/10.1007/s42770-023-01108-w

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  • DOI: https://doi.org/10.1007/s42770-023-01108-w

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