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A functional sweetener, difructose anhydride IV (DFA IV), is enzymatically produced from sucrose via levan by levansucrase (LSRase) followed by levan fructotransferase (LFTase). Here, we have demonstrated a consolidated production system for the direct conversion of DFA IV from sucrose using the co-culture of two recombinant yeast strains secreting LSRase from  Bacillus subtilis  and LFTase from  Arthrobacter ureafaciens , respectively. To ensure secretory production of the enzymes, target protein-specific translational fusion partners (TFP) were employed, and the selected strains produced 3.8 U/mL of LSRase and 16.0 U/mL LFTase activity into the fermentation broth. To optimise the direct production, sucrose concentration and cell ratios were investigated. In the optimised conditions, 64.3 g/L crude DFA IV was directly produced from 244.7 g/L sucrose using co-fermentation of recombinant yeasts. These results promise an efficient production titre, yield, and DFA IV productivity in an industrially applicable method.

Direct Production of Difructose Anhydride IV from Sucrose by Co-fermentation of Recombinant Yeasts