Fabricating the toolbox of MNPs, engineered enzymes and hybrid porous particles for the involved one-pot reactions, and the development of lab and pilot scale Alternating Magnetic Field (AMF)-bioreactors.
Turning energy transfer processes at the enzyme/MNP interphase by magnetic heating
he aim is to create a magnetic nanoparticles (MNP’s) toolbox with a wide range of AMF-tuned heating profiles. Their long-term stability under operation conditions (leakage of metal ions & toxicity) will be assessed.
Main objectives include:
- the modulation of MNP’s heating power by tuning several magnetic coupling parameters to obtain high temperature gradients at the enzymes surroundings regarding the bulk,
- to match the local temperature magnetically induced at the enzyme/MNPs interphase with each enzyme Topt;
- to ensure on demand or simultaneous ON/OFF thermal switching of different enzyme/MNPs conjugates.
Enzymes and MNPs surface engineering
In this work package, target enzymes with aminoacid clusters at different regions of their surface will be engineered to explore immobilization features that could affect MNP’s heat transfer. The nature, number and position of the aminoacids forming those clusters will be varied. Besides, complementary reactive chemical groups will be introduced at the surface of the different MNPs.
The aim is to find the best immobilization parameters that allow the optimum MNP’s heat transfer.
Development of hybrid composites for efficient localized heating
The goal of WP3 is the integration of optimal enzyme/MNPs combinations within porous microparticles without altering the optimal parameters already found to maximize heating transfer efficiency. This will be done with the example of a model two-enzyme cascade, consisting of a thermophilic (TOpt= 70 °C) and a mesophilic (TOpt= 37 °C) enzyme.
Development of AMF applicators for lab & pilot scale bioreactors
A set of novel AMF applicators will be developed in order to be able to activate multi-enzyme processes simultaneously or on demand. This will involve the design and prototyping of customized batch and flow lab-scale AMF-bioreactors operating with commercially available AMF technology environment on the one hand and novel AMF-devices for pilot scale reactors on the other.
Implementing challenging and industrially relevant one-pot enzymatic cascades
Simultaneous activation of one-pot multi-enzyme processes by AMF
AMF-tuned multi-enzyme cascades for conversion of sucroseinto α-glucosyl-2-glycerate and/or cellobiose/cello-oligosaccharides will be assembled. This would enable the operation of simultaneous enzymatic reactions with unpaired optimal temperatures through temperature-compartmentalized biotransformations within the same vessel.
On-demand activation of one-pot multi-enzyme processes by AMF
Assembling AMF-regulatable multi-enzyme cascades for the synthesis of tetrahydroisoquinolines in one-pot. The project will enable on demand activity regulation of each enzymatic step (ON/OFF) through temperature- compartmentalized biotransformation within the same vessel (e.g. by using a sequential reaction mode) to avoid undesired cross-reactivity.
Management, dissemination and exploitation
HOTZYMES ́s coordination, monitoring, dissemination and exploitation (IP protection) will be envisaged in WP7. Workshops focused on a specific topic will be organized to favour cross-fertilization between the consortium members. The results of HOTZYMES will be disseminated with posters and presentations at conferences, fairs and to the public and as well via articles in journals, press releases, social media, workshops and citizen science events. For exploitation purposes, a special event at the European Summit of Industrial Bitechnology (ESIB) will be held together with industrial partners. A Roadmap for future steps towards reaching HOTZYMES vision after the FET Open grant period will be set up.