Simultaneous enzymatic synthesis of gluconic acid and sorbitol. Continuous process development using glucose-fructose oxidoreductase from Zymomonas mobilis.

The production of sorbitol and gluconic acid by isolated glucose-fructose oxidoreductase (GFOR) from Zymomonas mobilis has been studied in a convective, 100-mL loop reactor with tangential ultrafiltration. Using a dilution rate of 0.04/h and 5 kU/L GFOR, substrate conversion (3 M sugar) in a single stage was >85%, and productivities of 126 g sorbitol/(L x d) were obtained. At a constant recycle rate (3/min) and a membrane area of 50 cm2, the dilution rates (and thus productivities) were however limited by a more than 30-fold reduction of the permeate flow in the presence of high sugar and protein concentrations (5 g/L). Protein was added, together with 10 mM dithiothreitol, to improve the stability of GFOR during substrate turnover and crossflow filtration, thus leading to a stable operation of the enzyme reactor for at least 5 d.