Abstract
Lipase B from Candida antarctica (CALB) was immobilized on octyl agarose (OC) and physically modified with polyethyleneimine (PEI) in order to confer a strong ion exchange character to the enzyme and thus enable the immobilization of other enzymes on its surface. The enzyme activity was fully maintained during the coating and the thermal stability was marginally improved. The enzyme release from the support by incubation in the non-ionic detergent Triton X-100 was more difficult after the PEI-coating, suggesting that some intermolecular physical crosslinking had occurred, making this desorption more difficult. Thermal stability was marginally improved, but the stability of the OCCALB-PEI was significantly better than that of OCCALB during inactivation in mixtures of aqueous buffer and organic cosolvents. SDS-PAGE analysis of the inactivated biocatalyst showed the OCCALB released some enzyme to the medium during inactivation, and this was partially prevented by coating with PEI. This effect was obtained without preventing the possibility of reuse of the support by incubation in 2% ionic detergents. That way, this modified CALB not only has a strong anion exchange nature, while maintaining the activity, but it also shows improved stability under diverse reaction conditions without affecting the reversibility of the immobilization.
Highlights
Lipases are the most used enzymes in biocatalysis due to their good activity, specificity, selectivity and robustness in a variety of reaction media [1,2,3]
On octyl agarose on octyl agarose to fail to produce the usual increase on enzyme activity found with other lipases [6]
The coating of octyl-CALB with PEI following the protocol successfully applied to enzyme co-immobilization has no negative effects on enzyme activity and has positive effects on enzyme stability, mainly in mixtures of organic cosolvents/aqueous buffers, even if this protocol was not optimized for this purpose
Summary
Lipases are the most used enzymes in biocatalysis due to their good activity, specificity, selectivity and robustness in a variety of reaction media [1,2,3]. In order to use them as industrial biocatalysts, immobilization is required in many instances in order to facilitate their reuse [10,11,12,13]. This step should be used to improve other enzyme features. The focus is on enzyme stability, but other enzyme properties may be improved, such as purity, activity, selectivity or specificity [14,15,16,17,18].
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