Solid-binding peptides (SBPs) are short amino acid sequences that act as molecular linkers to direct the orientated immobilisation of biomolecules onto solid matrices. Silica-based materials are suitable matrices for enzyme immobilisation in industrial processes. We have exploited the property a SBP that binds to materials that contain silica and have constructed a library of functional fusion proteins displaying binding affinity to this material whilst retaining high levels of enzyme activity. Cell-free synthetic biology circumvents many of the limitations encountered by in vivo synthetic biology by operating without the constraints of a cell. It provides a diverse range of new molecular tools that allow biocatalytic reaction systems to be engineered in a highly flexible manner. As a result, it offers higher substrate and enzyme loading and the facile optimisation of enzyme ratios. Some of the challenges of this approach include costly enzyme preparation, biocatalyst stability, and the need for constant supplementation with co-factors. To overcome these challenges, we present a molecular toolbox that facilitates the construction of biocatalytic modules with predefined functions and catalytic properties. The toolbox is comprised of three interchangeable building blocks: (a) low-cost inorganic matrices (e.g., silica, zeolite), (b) matrix-specific SBPs and (c) thermostable enzymes. The rational combination of these building blocks allows for flexibility and a ‘pick and mix’ and “re-use” approach with multiple biocatalytic modules available for the assembly of natural and non-natural pathways. Individual immobilised enzymes can be rationally combined to assemble recyclable and product-specific reactions. We present preliminary results on the introduction of this approach for the construction of two synthetic pathways for the conversion of organic waste. The pathway assembly process allows for rapid initial proof of concept evaluation and for the understanding of the parameters for a synthetic pathway, which would be very labour and time intensive by the in vivo approach. A/Prof Anwar Sunna Department of Chemistry and Biomolecular Sciences Faculty of Science and Engineering Macquarie University, NSW 2109, Australia T: +61 2 9850 4220 | F: +61 2 9850 8313 E: anwar.sunna@mq.edu.au<mailto:anwar.sunna@mq.edu.au> | mq.edu.au<http://mq.edu.au/> [Macquarie University]<http://mq.edu.au/> [1496386160177_Signature-Conference_WEB2.jpg]<http://www.synbioaustralasia.org/sba2017> CRICOS Provider Number 00002J. Think before you print. Please consider the environment before printing this email. This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message are those of the individual sender, and are not necessarily the views of Macquarie University.