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Separation and Immobilization of Lipase from Penicillium simplicissimum by Selective Adsorption on Hydrophobic Supports

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Abstract

Lipases are an enzyme class of a great importance as biocatalysts applied to organic chemistry. However, it is still necessary to search for new enzymes with special characteristics such as good stability towards high temperatures, organic solvents, and high stereoselectivity presence. The present work’s aim was to immobilize the lipases pool produced by Penicillium simplissicimum, a filamentous fungi strain isolated from Brazilian babassu cake residue. P. simplissicimum lipases were separated into three different fractions using selective adsorption method on different hydrophobic supports (butyl-, phenyl-, and octyl-agarose) at low ionic strength. After immobilization, it was observed that these fractions’ hyperactivation is in the range of 131% to 1133%. This phenomenon probably occurs due to enzyme open form stabilization when immobilized onto hydrophobic supports. Those fractions showed different thermal stability, specificity, and enantioselectivity towards some substrates. Enantiomeric ratio for the hydrolysis of (R,S) 2-O-butyryl-2-phenylacetic acid ranged from 1 to 7.9 for different immobilized P. simplissicimum lipase fractions. Asymmetry factor for diethyl 2-phenylmalonate hydrolysis ranged from 11.8 to 16.4 according to the immobilized P. simplissicimum lipase fractions. Those results showed that sequential adsorption methodology was an efficient strategy to obtain new biocatalysts with different enantioselectivity degrees, thermostability, and specificity prepared with a crude extract produced by a simple and low-cost technology.

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Acknowledgments

Financial support was gratefully received from PETROBRAS, FUJB, FAPERJ and CAPES.

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Authors and Affiliations

  1. Instituto de Química, Centro de Tecnologia, Universidade Federal do Rio de Janeiro, lab. 549-1, Ilha do Fundão, CEP 21945-970, Rio de Janeiro, RJ, Brazil

    Aline G. Cunha, Melissa L. E. Gutarra, Rodrigo V. Almeida, Lúcia M. C. Paiva & Denise M. G. Freire

  2. Departamento de Microbiología, Instituto de Fermentaciones Industrial, CSIC, Juan de la Cierva 3, 28006, Madrid, Spain

    Gloria Fernández-Lorente

  3. Centro de Pesquisa e Desenvolvimento Leopoldo Américo Miguez de Mello (CENPES), Petrobras, Brazil

    Juliana V. Bevilaqua

  4. Department of Biocatalysis, Institute of Catalysis, CSIC, Campus UAM, Cantoblanco, 28049, Madrid, Spain

    Roberto Fernández-Lafuente & Jose M. Guisán

Authors
  1. Aline G. Cunha
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  2. Gloria Fernández-Lorente
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  3. Melissa L. E. Gutarra
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  4. Juliana V. Bevilaqua
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  5. Rodrigo V. Almeida
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  6. Lúcia M. C. Paiva
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  7. Roberto Fernández-Lafuente
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  8. Jose M. Guisán
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  9. Denise M. G. Freire
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Correspondence to Denise M. G. Freire.

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Cunha, A.G., Fernández-Lorente, G., Gutarra, M.L.E. et al. Separation and Immobilization of Lipase from Penicillium simplicissimum by Selective Adsorption on Hydrophobic Supports. Appl Biochem Biotechnol 156, 133–145 (2009). https://doi.org/10.1007/s12010-008-8425-7

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  • DOI: https://doi.org/10.1007/s12010-008-8425-7

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