Applied Biochemistry and Biotechnology

, Volume 91, Issue 1–9, pp 739–752 | Cite as

Kinetic studies of lipase from Candida rugosa

A comparative study between free and immobilized enzyme onto porous chitosan beads
  • Ernandes B. Pereira
  • Heizir F. De Castro
  • Flávio F. De Moraes
  • Gisella M. ZaninEmail author


The search for an in expensive support has motivated our group to undertake this work dealing with the use of chitosan as matrix for immobilizing lipase. In addition to its low cost, chitosan has several advantages for use as a support, including its lack of toxicity and chemical reactivity, allowing easy fixation of enzymes. In this article, we describe the immobilization of Canada rugosa lipase onto porous chitosan beads for the enzymatic hydrolysis of oliveoil. The binding of the lipase onto the support was performed by physicalad sorption using hexane as the dispersion medium. A comparativestudy between free and immobilized lipase was conducted in terms of pH, temperature, and thermal stability. A slightly lower value for optimum pH (6.0) was found for the immobilized form in comparison with that attained for the soluble lipase (7.0). The optimum reaction temperature shifted from 37°C for the free lipase to 50°C for the chitosan lipase. The patterns of heat stability indicated that the immobilization process tends to stabilize the enzyme. The half-life of the soluble free lipase at 55°C was equal to 0.71 h (K d=0.98 h−1), whereas for the immobilized lipase it was 1.10 h (K d=0.63 h−1). Kinetics was tested at 37°C following the hydrolysis of olive oil and obeys the Michaelis-Menten type of rate equation. The K m was 0.15 mM and the V max was 51 μmol/(min·mg), which were lower than for free lipase, suggesting that the apparent affinity toward the substrate changes and that the activity of the immobilized lipase decreases during the course of immobilization.

Index Entries

Lipase immobilization chitosan physical adsorption characterization hydrolysis 


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Copyright information

© Humana Press Inc 2001

Authors and Affiliations

  • Ernandes B. Pereira
    • 1
  • Heizir F. De Castro
    • 2
  • Flávio F. De Moraes
    • 1
  • Gisella M. Zanin
    • 1
    Email author
  1. 1.Department of Chemical EngineeringMaringá State UniversityMaringá-PRBrazil
  2. 2.Department of Chemical EngineeringFaculty of Chemical Engineering of LorenalLorena-SPBrazil

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