Abstract
In this work, we propose an adaptation strategy for Scheffersomyces stipitis NRRL Y-7124 to increase its tolerance to inhibitors of hemicellulosic hydrolysates in the production of bioethanol. Sugarcane bagasse pretreated with 0.5% (v/v) of sulfuric acid at 140 °C for 15 min was used as feedstock. The microorganism was subjected to a cell recycle batch fermentation system with increased inhibitors pressure, characterized by five consecutive batches with 33% of hydrolysate, followed by five subsequent batches with 67% of hydrolysate. The remaining composition of the culture media for each sequential batch (necessary to complete 100% of carbohydrate content) was synthetic substrate. Finally, the strain was used in a fed-batch fermentation with 100% of hydrolysate, resulting in an adapted strain of S. stipitis at the end. In order to validate the adaptation strategy, the fermentative performances of the adapted and non-adapted strains were compared under different acetic acid concentrations using synthetic substrate. 0.7 g/L of acetic acid significantly affected the fermentative performance of the non-adapted strain, while for the adapted strain, this value was 3.7-fold higher (2.6 g/L), confirming the increased tolerance. Through proteomic analysis, genes related to the defense mechanisms of S. stipitis against oxidative stress due to the presence of inhibitory compounds were identified, which plays a key role on the comprehension of the cell adaptation metabolism.
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Acknowledgements
The authors acknowledge the Brazilian Center of Research in Energy and Materials (CNPEM), Brazilian Biorenewables National Laboratory (LNBR), Brazilian Biosciences National Laboratory (LNBio), Coordination of Improvement of Higher Level Personnel (CAPES), and Foundation for Research Support of the State of São Paulo (FAPESP) (process numbers 2014/01135-0, 2015/20630-4, 2016/14567-0, and 2017/04997-0) for financial support, assistance regarding the use of facilities, the sharing of expertise, and the opportunity to develop this work.
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The authors acknowledge the Brazilian Center of Research in Energy and Materials (CNPEM), Brazilian Biorenewables National Laboratory (LNBR), Brazilian Biosciences National Laboratory (LNBio), Coordination of Improvement of Higher Level Personnel (CAPES), and Foundation for Research Support of the State of São Paulo (FAPESP) (process numbers 2014/01135-0, 2015/20630-4, 2016/14567-0, and 2017/04997-0) for financial support
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Biazi, L.E., Santos, S.C., Kaupert Neto, A.A. et al. Adaptation Strategy to Increase the Tolerance of Scheffersomyces stipitis NRRL Y-7124 to Inhibitors of Sugarcane Bagasse Hemicellulosic Hydrolysate Through Comparative Studies of Proteomics and Fermentation. Bioenerg. Res. 15, 479–492 (2022). https://doi.org/10.1007/s12155-021-10267-3
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DOI: https://doi.org/10.1007/s12155-021-10267-3