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Applied Biochemistry and Biotechnology

, Volume 146, Issue 1–3, pp 49–56 | Cite as

Immobilization of Yarrowia lipolytica Lipase—a Comparison of Stability of Physical Adsorption and Covalent Attachment Techniques

  • Aline G. Cunha
  • Gloria Fernández-Lorente
  • Juliana V. Bevilaqua
  • Jacqueline Destain
  • Lúcia M. C. Paiva
  • Denise M. G. Freire
  • Roberto Fernández-Lafuente
  • Jose M. Guisán
Article

Abstract

Lipase immobilization offers unique advantages in terms of better process control, enhanced stability, predictable decay rates and improved economics. This work evaluated the immobilization of a highly active Yarrowia lipolytica lipase (YLL) by physical adsorption and covalent attachment. The enzyme was adsorbed on octyl–agarose and octadecyl–sepabeads supports by hydrophobic adsorption at low ionic strength and on MANAE–agarose support by ionic adsorption. CNBr–agarose was used as support for the covalent attachment immobilization. Immobilization yields of 71, 90 and 97% were obtained when Y. lipolytica lipase was immobilized into octyl–agarose, octadecyl–sepabeads and MANAE–agarose, respectively. However, the activity retention was lower (34% for octyl–agarose, 50% for octadecyl–sepabeads and 61% for MANAE–agarose), indicating that the immobilized lipase lost activity during immobilization procedures. Furthermore, immobilization by covalent attachment led to complete enzyme inactivation. Thermal deactivation was studied at a temperature range from 25 to 45°C and pH varying from 5.0 to 9.0 and revealed that the hydrophobic adsorption on octadecyl–sepabeads produced an appreciable stabilization of the biocatalyst. The octadecyl–sepabeads biocatalyst was almost tenfold more stable than free lipase, and its thermal deactivation profile was also modified. On the other hand, the Y. lipolytica lipase immobilized on octyl–agarose and MANAE–agarose supports presented low stability, even less than the free enzyme.

Keywords

Immobilized lipase Y. lipolytica Biocatalysis Glutaraldehyde Hydrophobic supports 

Notes

Acknowledgements

Financial support was gratefully received from PETROBRÁS, FUJB, FAPERJ and CAPES. The authors are also grateful to Prof Rodrigo Volcan Almeida for his contribution.

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Copyright information

© Humana Press Inc. 2007

Authors and Affiliations

  • Aline G. Cunha
    • 1
  • Gloria Fernández-Lorente
    • 2
  • Juliana V. Bevilaqua
    • 3
  • Jacqueline Destain
    • 4
  • Lúcia M. C. Paiva
    • 1
  • Denise M. G. Freire
    • 1
  • Roberto Fernández-Lafuente
    • 2
  • Jose M. Guisán
    • 2
  1. 1.Instituto de Química, Universidade Federal do Rio de Janeiro, Centro de Tecnologia (CT)Ilha do FundãoBrazil
  2. 2.Department of BiocatalysisInstitute of CatalysisCantoblancoSpain
  3. 3.Centro de Pesquisa e Desenvolvimento Leopoldo Américo Miguez de Mello (CENPES)PetrobrasBrazil
  4. 4.Centre Wallon de Biologie Industrielle, Faculté universitaire des Sciences agronomiquesGemblouxBelgium

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