Reactivation of covalently immobilized lipase from Thermomyces lanuginosus

Rafael C. Rodrigues, Cesar A. Godoy, Marco Filice, Juan M. Bolivar, Armand Palau-Ors, Jesus M. Garcia-Vargas, Oscar Romero, Lorena Wilson, Marco A.Z. Ayub, Roberto Fernandez-Lafuente, Jose M. Guisan

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Lipase from Thermomyces lanuginosus (TLL) immobilized on cyanogen bromide agarose (CNBr) may be fully inactivated when incubated in saturated solutions of guanidine. When this inactivated enzyme is re-incubated in aqueous medium, 20% of the activity may be recovered for several cycles. However, if the activity was determined in the presence of a detergent (CTAB, an activator of this enzyme), 100% of the initial activity in the presence of detergent was recovered. The enzyme was also inactivated in the presence of organic solvents and at high temperatures. Inactivations were more rapid when the activity was determined in absence of detergent. In both cases, some activity could be recovered just by incubation under mild conditions, and this increase was higher if the activity measurements were performed in the presence of CTAB. These results suggested that the opening of the lipase could be a critical step in the inactivation or reactivation of immobilized TLL. In inactivations in the presence of solvents, 100% of activity could be recovered during several cycles, while in thermal inactivations, the recovered activity decreased in each inactivation-reactivation cycle. The incubation of the enzyme inactivated by temperature in guanidine improved the results, but still 100% could not be achieved during several cycles even measured in the presence of CTAB. Thus, the simple incubation of the partially or fully inactivated enzyme under mild conditions permitted to recover some activity (enhancing the half life of the biocatalysts), even in thermal inactivations.

Original languageEnglish
Pages (from-to)641-646
Number of pages6
JournalProcess Biochemistry
Issue number6
StatePublished - Jun 2009


  • Enzyme inactivation
  • Enzyme reactivation
  • Immobilized enzymes
  • Operational stabilization
  • Unfolding-refolding


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