Abstract
Lipase activity (337 U/g dry weight of cell debris) was detected in cell debris after ultrasound treatment of Yarrowia lipolytica cells cultivated in residual frying palm oil. It is a naturally immobilized lipase with protein content of 47%, herein called LipImDebri. This immobilized biocatalyst presents low hydrophobicity (8%), that can be increased adjusting pH and buffer type. Despite apparent intact cells, electron microscopy showed a shapeless and flat surface for LipImDebri and optical microscopy revealed no cell viability. Besides, an inferior mean diameter (3.4 mm) in relation to whole cells reveals structure modification. A high negative zeta potential value (− 33.86 mV) for pH 6 and 25 °C suggests that LipImDebri is a stable suspension in aqueous solution. Fourier Transform Infrared Spectra (FTIR) expose differences between LipImDebri and extracellular lipase extract signaling a physical interaction between enzyme and cell debris, which is possibly the reason for the high thermostability (kd = 0.246 h−1; t1/2 = 2.82 h at 50 °C, pH 7.0). A good adjustment of LipImDebri kinetic data with Hill equation (R2 = 0.95) exposes an allosteric behavior related to the presence of more than one lipase isoform. These features reveal that LipImDebri can be a good catalyst for industrial applications.
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Acknowledgements
The authors acknowledge the DRX analysis performed by Nuclear Instrumentation Laboratory and SEM analysis performed by Scanning Electronic Microscopy Lab, both from Universidade Estadual do Rio de Janeiro (UERJ), Brazil. The authors acknowledge the analysis (FTIR, Zeta potential) provided by Prof. Priscilla Vanessa Finotelli (Universidade Federal do Rio de Janeiro) that contributed to a better discussion of the results of the present study.
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The financial support of Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ—grant number E-26/202.870/2015 BOLSA), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES—001/Bolsa) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq—Bolsa).
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Conceptualization: PFFA, KAS; Data curation: PFFA; Formal analysis: JLF; Funding acquisition: PFFA; Investigation: ACBP, JLF; Methodology: JLF, PVF, PFFA; Project administration: PFFA; Resources: PFFA; Software: JLF, PFFA; Supervision: PFFA, EA, KAS; Validation: JLF; Visualization: JLF, PFFA; Roles/Writing—original draft: JLF; Writing—review and editing: PFFA, PVF, EA, KAS.
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Fraga, J.L., da Penha, A.C.B., Akil, E. et al. Catalytic and physical features of a naturally immobilized Yarrowia lipolytica lipase in cell debris (LipImDebri) displaying high thermostability. 3 Biotech 10, 454 (2020). https://doi.org/10.1007/s13205-020-02444-6
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DOI: https://doi.org/10.1007/s13205-020-02444-6