Abstract

This study investigates the immobilisation of Pseudomonas stutzeri lipase and its reactivation as a strategy to extend the biocatalyst's operational life after partial inactivation. Two immobilised catalysts were prepared using glyoxyl agarose and heterofunctional agarose supports. These biocatalysts were inactivated in 90% (v/v) 1,4-dioxane at 25 degrees C until reaching approximately 60% residual activity. The reactivation process evaluated the effects of temperature and the presence of Triton X-100 detergent. The most effective reactivation in aqueous medium was achieved with the glyoxyl agarose biocatalyst, recovering 100% of its initial activity in all tested cases. The results indicate that a higher number of covalent bonds between the enzyme and the support facilitates the refolding of the biocatalyst into its active conformation. Although increasing the temperature positively affected the reactivation rate, the addition of Triton X-100 to the medium resulted in the highest reactivation rate and activity recovery for both biocatalysts. Selecting an appropriate immobilisation strategy not only improves enzyme stability but also enables the structural reactivation of partially inactivated enzymes, significantly extending their useful life under process conditions.