Real-time NMR monitoring of spatially segregated enzymatic reactions in multilayered hydrogel assemblies

• Main Authors: Nordin, Nurdiana, Bordonali, Lorenzo, Davoodi, Hossein, Ratnawati, Novindi Dwi, Gygli, Gudrun, Korvink, Jan G., Badilita, Vlad, MacKinnon, Neil
• Format: Article
• Published: Wiley-V C H Verlag GMBH 2021
• Subjects: QD Chemistry
• Online Access: http://eprints.um.edu.my/34001/
• Institution: Universiti Malaya

Compartmentalized chemical reactions at the microscale are important in biotechnology, yet monitoring the molecular content at these small scales is challenging. To address this challenge, we integrate a compact, reconfigurable reaction cell featuring electrochemical functionality with high-resolution NMR spectroscopy. We demonstrate the operation of this system by monitoring the activity of enzymes immobilized in chemically distinct layers within a multi-layered chitosan hydrogel assembly. As a benchmark, we observed the parallel activities of urease (Urs), catalase (Cat), and glucose oxidase (GOx) by monitoring reagent and product concentrations in real-time. Simultaneous monitoring of an independent enzymatic process (Urs) together with a cooperative process (GOx + Cat) was achieved, with chemical conversion modulation of the GOx + Cat process demonstrated by varying the order in which the hydrogel was assembled.