Design of the Enzyme–Carrier Interface to Overcome the O<sub>2</sub> and NADH Mass Transfer Limitations of an Immobilized Flavin Oxidase
Ana I. Benítez‐Mateos, Christina Huber, Bernd Nidetzky, Juan M. Bolívar, Fernando López‐Gallego
Abstract
. Furthermore, the co-immobilization of NADH and NOX within the tuned surface of porous microbeads increases the effective concentration of NADH in the surroundings of the enzyme. As a result, the optimal spatial organization of NOX and its confinement with NADH allow a 100% recovery of the activity of the soluble enzyme upon the immobilization process. By engineering these variables, we increase the NADH oxidation activity of the heterogeneous biocatalyst by up to 650% compared to NOX immobilized under suboptimal conditions. In conclusion, this work highlights the rational design and engineering of the enzyme-carrier interface to maximize the efficiency of heterogeneous biocatalysts.