The research of simple
and fast enzyme immobilization methods,
preserving the enzyme activity and improving the thermal stability,
is in the spotlight. The objective of this work is to develop a β-galactosidase
immobilization one-pot route, combining the silica sol–gel
encapsulation (SSGE) process with a metal chelation strategy by using
chitosan and Ca2+, Zn2+, or Cu2+ cations.
The results show that the presence of cations does not affect the
encapsulation efficiency (81%) and has positive effects on the maximum
catalytic potential, especially at 60 °C and in the presence
of Ca2+ ions (MPC = 2203). They enhance the biocatalyst
thermal stability and promote hyperactivation with respect to the
soluble enzyme at 60 °C (1.6 times higher MPC). The biocatalyst
prepared with Zn2+ ions exhibits also thermal hyperactivation
in the first 30 min of heating (1.3 times more residual activity),
but the enzyme is not stabilized (0.9 times lower MPC); also, the
presence of Cu2+ ions does not promote hyperactivation
or stabilization of the enzyme (0.3 times lower MPC) at this high
temperature. These facts are reflected in the hydrolytic and transgalactosylation
activities of the enzyme (33.6–57.4% total lactose conversion),
higher than that reported with analogue biocatalysts. The physicochemical
characterization of the obtained solid biocatalysts by SEM, TEM, XRF,
and XPS indicates that chitosan–metal chelation has an important
role in the encapsulation process and that a low metal degree incorporation
(8.85 ppm of Ca2+) on the solid biocatalyst favors the
thermal hyperactivation and stabilization of the evaluated β-galactosidase.
This work contributes to the understanding of the SSGE process mediated
by chitosan–metal chelates, which is a simple and fast one-pot
immobilization strategy.