Abstract
The recombinant invertase INVB (re-INVB) from Zymomonas mobilis was immobilized on microbeads of Nylon-6, by means of covalent bonding. The enzyme was strongly and successfully bound to the support. The activity of the free and immobilized enzyme was determined, using 10% (w/v) sucrose, at a temperature ranging between 15 and 60 °C and a pH ranging between 3.5 and 7. The optimal pH and temperature for the immobilized enzyme were 5.5 and 25 °C, respectively. Immobilization of re-INVB on Nylon-6 showed no significant change in the optimal pH, but a difference in the optimal temperature was evident, as that for the free enzyme was shown to be 40 °C. The values for kinetic parameters were determined as: 984 and 98 mM for \( K_{m}^{\text{app}} \) of immobilized and free re-INVB, respectively. \( K_{\text{cat}}^{\text{app}} \) values for immobilized and free enzymes were 6.1 × 102 and 1.2 × 104 s−1, respectively, and immobilized re-INVB showed \( V_{\max }^{\text{app}} \) of 158.73 μmol h min−1 mg−1. Immobilization of re-INVB on Nylon-6 enhanced the thermostability of the enzyme by 50% at 30 °C and 70% at 40 °C, when compared to the free enzyme. The immobilization system reported here may have future biotechnological applications, owing to the simplicity of the immobilization technique, the strong binding of re-INVB to the support and the effective thermostability of the enzyme.
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Acknowledgments
The research was funded by the Centro de Investigación y de Estudios Avanzados (CINVESTAV-IPN), México. V.V.B gratefully acknowledges a scholarship from the Consejo Nacional de Ciencia y Tecnología (Conacyt), México.
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Vallejo-Becerra, V., Vásquez-Bahena, J.M., Santiago-Hernández, J.A. et al. Immobilization of the recombinant invertase INVB from Zymomonas mobilis on Nylon-6. J Ind Microbiol Biotechnol 35, 1289–1295 (2008). https://doi.org/10.1007/s10295-008-0426-6
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DOI: https://doi.org/10.1007/s10295-008-0426-6