Conclusions We developed a new method for FAD-mediated flavoenzyme immobilization. It yielded fully active and stable immobilized bio-catalysts. For this method, agarose was covalently functionalizedwith a synthetic FAD cofactor which could be used for an efficientand quick reconstitution and immobilization of apo PAMO and apoPTDH-PAMO. The immobilized enzymes restored full activity, couldbe reused, and displayed a higher thermostability than the enzymein solution. The FAD-agarose was shown to be a robust carrier, recy-clable for multiple immobilizations. The presented method opens up new avenues for the tunable and gentle immobilization of other flavoenzymes