Abstract
New efficient protein purification methods need to be developed to overcome product loss during purification and to lower its final price. Magnetic adsorbents emerge as a potential support which can easily separate target biomolecules from crude biological medium. The hydrophobic adsorption of recombinant lipase from Bacillus thermocatenulatus (BTL-2) in novel silica magnetic microparticles (SMMp) was investigated aiming to directly recover it from cell lysate. The adsorbent surface of SMMp was modified by activation with octyl groups (25%–100%). Adsorptions on silica-octyl (non-magnetic) were also performed to understand the process better. Using centrifuged and dialyzed enzyme, under low ionic strength, the highest enzyme adsorption (92%) and desorption yields (50%) were obtained using SMMp-octyl-75% and SMMp-octyl-25%, respectively, due to simultaneous hydrophobic and ionic interaction between charged silanol groups and the enzyme. Direct enzyme adsorption from cell lysates under high ionic strength conditions led to a high yield of enzyme adsorption with 100% desorption using SMMp-octyl-25%. Therefore, the use of SMMp-octyl allows for a simple and highly efficient BTL-2 recovery by a one-step purification technique that can even be extended to other applications.
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Acknowledgements
The authors are grateful to the Laboratory of Biocatalysis (ICP-CSIC, Madrid, Spain) for the donation of recombinant cells of E. coli, and the São Paulo Research Foundation (FAPESP grants #2008/56246-0 and 2016/10636-8), the Brazilian National Council for Scientific and Technological Development (CNPq) and the Coordenação de Aperfeiçoamento de Pessoal de NÃvel Superior (CAPES) for the financial support.
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This work was supported by the São Paulo Research Foundation (FAPESP), grants #2008/56246-0 and 2016/10636-8, and the Brazilian National Council for Scientific and Technological Development (CNPq) for their financial support. This work was in part financed by the Coordenação de Aperfeiçoamento de Pessoal de NÃvel Superior, Brazil (CAPES) – Finance code 001.
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All authors contributed to the study conception and design. LAL performed all experiments with the support of WK. All authors discussed and analysed the data. The first draft of the manuscript was written by LAL and manuscript review and editing were carried out by TSM and TP. RLCG revised the manuscript and supervised the work development. All authors read and approved the final manuscript.
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Lopes, L.A., Kopp, W., Milessi, T.S. et al. Direct recovery of intracellular lipase from cell lysate by adsorption on silica magnetic microparticles activated with Octyl groups. Braz. J. Chem. Eng. 39, 789–801 (2022). https://doi.org/10.1007/s43153-021-00190-7
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DOI: https://doi.org/10.1007/s43153-021-00190-7