Partners involved: UCL (WP leader), TUG (Nidetzky), UL, Micronit, ix-factory, ChipShop, DTU, TUB, nanoMyP.
WP7 will establish microfluidic chemo-biocatalytic reaction systems that enable rapid characterisation of biosynthetic pathways and chemo-enzymatic conversions and that can effectively compete with automated micro well approaches in terms of through-put, mass fabrication and translation of results to larger reactor scales. They will be integrated with early-stage downstream operation steps, optimized based on developed modelling methods and benchmarked against existing larger scale data.
The application of microfluidic enzyme reactors for early process development and their potential for introduction into industrial-scale synthesis needs to be further exploited in several areas: Firstly, cascading chemical and multi-enzymatic reaction process steps will facilitate the rapid development of novel routes for the synthesis of complex chiral precursors for the next generation of pharmaceuticals, yet to date little work has been published in this area. Secondly, integration with downstream processing steps and modeling-based reactor optimization will further improve the applicability of these systems for industrial-scale synthesis. Finally, high-yielding immobilisation methods are required to expand the range of enzyme classes that can be used in microfluidic reactors and to maximise stability of the enzyme and amount that can be loaded. Various immobilisation strategies have been applied, by other groups and consortium partners. However, further research into robust, high-yielding and reversible immobilization methods is required, to achieve this goal, and thus facilitate the rapid screening against chemo-enzymatic process conditions.
Research plan
In the first year an immobilised microfluidic enzymatic reactor (IMER) will be fabricated for the rapid evaluation of kinetic parameters of single enzyme variants. For the integration of enzymatic and chemical process steps, a commercial microfluidic chemical reactors will also be employed and kinetic parameters of cycloaddition reactions, a representative, extensively used chemical transformation will be studied.
In the second year the conversion yields from different enzyme will be compared in immobilised and non-immobilised microfluidic reactors. Using the platform developed in the first year, the enzymatic reactions can be undertaken with different flow rates and substrate concentrations, which will enable determination of both the kinetic parameters and the conditions for high conversions. In interaction with WP8, pH sensors and other analytical methods such as optical nano-sensor particles for rapid detection of process conditions and product formation into the microfluidic reactor will be integrated.
In the third year, the technologies developed in previous years will be employed to create microfluidic platforms suitable for multi-step reactions and biotransformations with integrated product separation within microstructured devices.