Affinity directed assembly of multilayers of pectinase

Abstract

Numerous approaches to creating protein multilayers have been described for a variety of applications. In this work, the affinity of the enzyme pectinase towards a natural polysaccharide alginate has been exploited to create affinity directed enzyme multilayers on alginate beads. The biocatalyst has been characterized in terms of catalytic efficiency and kinetic parameters. Atomic force microscopy provided a good insight into multilayer formation at the molecular level. It was seen that enzyme layering was not uniform all over the bead. Thus, achieving high catalytic efficiency was an ‘average performance’ of the enzyme molecule, which was part of different layers. The results show that it is possible to create layered biocatalyst designs without using expensive lectins/antibodies for constructing affinity layers.

• Atomic force microscopy
• affinity directed protein assembly
• protein multilayers
• pectinase