Solid-binding peptides for immobilisation of thermostable enzymes to hydrolyse biomass polysaccharides

Andrew Care, Kerstin Petroll, Emily S. Y. Gibson, Peter L. Bergquist, Anwar Sunna

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)
16 Downloads (Pure)

Abstract

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.
Original languageEnglish
Article number29
Pages (from-to)1-16
Number of pages16
JournalBiotechnology for Biofuels
Volume10
Issue number1
DOIs
Publication statusPublished - 2 Feb 2017

Bibliographical note

Copyright the Author(s) 2017. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

Keywords

  • solid-binding peptide
  • thermostable enzymes
  • CLEAs
  • enzyme immobilisation
  • biocatalytic modules

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