Chemobiocatalytic Production of 5-Hydroxymethyl-2-furancarboxylic Acid from Sucrose by a Choline Chloride–Lactic Acid Deep Eutectic Solvent and a Novel <i>Gluconobacter oxidans</i> Strain

5-Hydroxymethyl-2-furancarboxylic acid (HMFCA), a critical polyester intermediate, faces an industrial scale-up challenge due to the high production cost using commercial 5-hydroxymethylfurfural (HMF) as a substrate. This study developed an efficient chemobiocatalytic route to synthesize HMFCA from inexpensive sucrose. A newly isolated <i>Gluconobacter oxidans</i> strain showed high HMF tolerance (246 mM) and completely converted HMF to HMFCA in 24 h with 99.3% yield (mol/mol). Meanwhile, the choline chloride-lactic acid deep eutectic solvent was found to efficiently convert sucrose to HMF, yielding 106.4 mM HMF (53.2% yield). The chemobiocatalytic potential of HMFCA production was evaluated using sucrose-derived crude HMF and <i>G. oxidans</i> whole-cell catalysis. Within 24 h, the crude HMF (64.8 mM) was completely consumed, and 63.5 mM HMFCA was produced. This integrated process achieved an overall HMFCA yield of approximately 52% based on the initial sucrose. These findings provide an efficient strategy for chemobiocatalytic conversion of inexpensive sucrose to high-value HMFCA.