Three intertwining effects guide the mode of action of chitin deacetylase de- and N-acetylation reactions.

Chitosans are promising multi-functional biomolecules for various applications whose performance is dependent on three key structural parameters, including the pattern of acetylation (PA). To date, chitin deacetylases (CDAs) are the only tool to control the PA of chitosan polymers via their specific mode of action during de- or N-acetylation. For a start, this review summarizes the current state of research on the classification of carbohydrate esterase 4 enzymes, the features in sequence and structure of CDAs, and the different PAs produced by different CDAs during de- or N-acetylation. In the main part, we introduce three effects that guide the mode of action of these enzymes: the already established subsite capping effect, the subsite occupation effect, and the subsite preference effect. We show how their interplay controls the PA of CDA products and describe their molecular basis. For one thing, this review aims to equip the reader with the knowledge to understand and analyze CDAs - including a guide for in silico and in vitro analyses. But more importantly, we intend to reform and extend the model explaining their mode of action on chitosans to facilitate a deeper understanding of these important enzymes for biology and biotechnology.