Solid-binding peptides for immobilisation of thermostable enzymes to hydrolyse biomass polysaccharides.
BACKGROUND
Solid-binding peptides (SBPs) bind strongly to a diverse range of solid materials without the need for any chemical reactions. They have been used mainly for the functionalisation of nanomaterials but little is known about their use for the immobilisation of thermostable enzymes and their feasibility in industrial-scale biocatalysis.


RESULTS
A silica-binding SBP sequence was fused genetically to three thermostable hemicellulases. The resulting enzymes were active after fusion and exhibited identical pH and temperature optima but differing thermostabilities when compared to their corresponding unmodified enzymes. The silica-binding peptide mediated the efficient immobilisation of each enzyme onto zeolite, demonstrating the construction of single enzyme biocatalytic modules. Cross-linked enzyme aggregates (CLEAs) of enzyme preparations either with or without zeolite immobilisation displayed greater activity retention during enzyme recycling than those of free enzymes (without silica-binding peptide) or zeolite-bound enzymes without any crosslinking. CLEA preparations comprising all three enzymes simultaneously immobilised onto zeolite enabled the formation of multiple enzyme biocatalytic modules which were shown to degrade several hemicellulosic substrates.


CONCLUSIONS
The current work introduced the construction of functional biocatalytic modules for the hydrolysis of simple and complex polysaccharides. This technology exploited a silica-binding SBP to mediate effectively the rapid and simple immobilisation of thermostable enzymes onto readily-available and inexpensive silica-based matrices. A conceptual application of biocatalytic modules consisting of single or multiple enzymes was validated by hydrolysing various hemicellulosic polysaccharides.
Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, Australia.|ARC Centre of Excellence for Nanoscale BioPhotonics (CNBP), Macquarie University, Sydney, Australia.
Andrew
A
Care
Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, Australia.
Kerstin
K
Petroll
Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, Australia.
Emily S Y
ESY
Gibson
Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, Australia.|Department of Molecular Medicine & Pathology, Medical School, University of Auckland, Auckland, New Zealand.|Biomolecular Discovery and Design Research Centre, Macquarie University, Sydney, Australia.
Peter L
PL
Bergquist
Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, Australia.|Biomolecular Discovery and Design Research Centre, Macquarie University, Sydney, Australia.
Anwar
0000-0002-1467-9937
A
Sunna
Biocatalytic modules; CLEAs; Enzyme immobilisation; Solid-binding peptide; Thermostable enzymes