This thesis discusses two major aspects involved in the transition of biocatalysis from simple batch operation to spatially and temporally compartmentalised continuous flow systems. To support this transition, engineering of enzymes and cofactors is essential. In this research, enzyme engineering for the preparation of immobilised enzymes was explored. Enzymes were genetically fused to non-covalent affinity tags and immobilised on low-cost carriers for application in flow assisted biocatalytic protocols. In addition, novel nicotinamide cofactor biomimetics that may allow cofactor tethering were prepared to support implementation of cost-effective cofactor-dependent redox biocatalytic methodologies.