Carbohydrate-active enzymes are accountable for the synthesis and degradation of glycosidic bonds among diverse carbohydrates. Fructosyl-transferases represent a subclass of these enzymes, employing sucrose as a substrate to generate fructooligosaccharides (FOS) and fructan polymers. This category primarily includes levansucrase (LS, EC 2.4.1.10), inulosucrase (IS, EC 2.4.1.9), and β-fructofuranosidase (Ffase, EC 3.2.1.26). These three enzymes possess a similar five-bladed β-propeller fold and employ an anomer-retaining reaction mechanism mediated by nucleophiles, transition state stabilizers, and general acids/bases. However, they exhibit distinct product profiles, characterized by variations in linkage specificity and molecular mass distribution. Consequently, this article comprehensively explores recent advancements in the catalytic characteristics, structural features, reaction mechanisms, and product specificity of levansucrase, inulosucrase, and β-fructofuranosidase (abbreviated as LS, IS, and Ffase, respectively). Furthermore, it discusses the potential for modifying catalytic properties and product specificity through structure-based design, which enables the rational production of custom fructan and FOS.

Ectomycorrhizal (ECM) symbiosis is a mutualistic interaction between certain soil fungi and fine roots of perennial plants, mainly forest trees, by which both partners become capable of efficiently colonising nutrient-limited environments. The success of this interaction is reflected in the dominance of ECM forest ecosystems in the Northern hemisphere. Apart from their economic importance (wood production), forest ecosystems are essential for large-scale carbon sequestration, leading to substantial reductions in anthropogenic CO(2) release. The biological function of ECM symbiosis is the exchange of fungus-derived mineral nutrients for plant-derived carbohydrates. Improved plant nutrition as a result of this interaction, however, has a price. Together with their fungal partner, root systems of ECM plants can receive about half of the photosynthetically fixed carbon. To enable such a strong carbohydrate sink, the monosaccharide uptake capacity and carbohydrate flux through glycolysis and intermediate carbohydrate storage pools (trehalose and/or mannitol) of mycorrhizal fungi is strongly increased at the plant-fungus interface. Apart from their function as a carbohydrate store, trehalose/mannitol are additionally considered to be involved in carbon allocation within the fungal colony. Dependent on the fungal species involved in the symbiosis, regulation and fine-tuning of fungal carbohydrate uptake and metabolism seems to be controlled either by developmental mechanisms or by the apoplastic sugar content. As a consequence of the increased carbohydrate demand in symbiosis, trees increase their photosynthetic capacity. In addition, host plants control and restrict carbohydrate flux towards their partner to avoid fungal parasitism. The mechanisms behind this phenomenon are still largely unknown but rates of local sucrose hydrolysis and hexose uptake by rhizodermal cells are thought to restrict fungal carbohydrate nutrition under certain conditions (e.g., reduced fungal nutrient export).