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Zeta potential measurement as a diagnostic tool in enzyme immobilisation
Ist Teil von
Colloids and surfaces, B, Biointerfaces, 2008-10, Vol.66 (1), p.39-44
Ort / Verlag
Netherlands: Elsevier B.V
Erscheinungsjahr
2008
Quelle
ScienceDirect
Beschreibungen/Notizen
The efficiency of binding during enzyme immobilisation does not only depend on the chemical properties of the enzyme and the matrix particle, but also on their surface potential. Zeta potential quantifies the electrostatic interactions between enzyme and matrix particles, and can therefore, be used as an indicator of the binding efficiency in the enzyme immobilisation studies. In order to establish a correlation between the zeta potential and the binding efficiency, we used CALA (
Candida antarctica A-type lipase) as a model protein for immobilisation on non-porous magnetic microparticles with epoxy (M-PVA E02), carboxy (M-PVA C12) and amine (M-PVA N12) terminations. We observed maximal binding of CALA onto the M-PVA N12 beads, due to the electrostatic attraction between negatively charged protein and carrier particles with slightly positive zeta potential. The binding of CALA was lower when M-PVA E02 beads were used, followed by M-PVA C12 beads. The decreasing binding efficiency was obviously the result of increasing electrostatic repulsion between the interaction partners. This could be correlated to the increasing negative zeta potential of the magnetic particles. Moreover, the medium of suspension of the particles also makes a significant difference. We found highest specific activity of the lipase immobilised on M-PVA E02 beads in a medium concentrated buffer (0.3
M). The results demonstrate a clear correlation between zeta potential and binding efficiency but no correlation between the bead related specific activity and the zeta potential. These findings are advocating the possibility of using the zeta potential as a diagnostic tool in enzyme immobilisation.