An enzymatic, NAD(H)-dependent process for the efficient production of D-ma
nnitol from D-fructose as one single product is described and optimized wit
h respect to productivity at high substrate conversion. Stereospecific redu
ction of D-fructose is catalyzed by recombinant mannitol dehydrogenase from
Pseudomonas fluorescens DSM 50106, overexpressed. in Escherichia coli. Reg
eneration of NADH is accomplished by formate dehydrogenase-mediated oxidati
on of formate into CO?, thus avoiding byproduct formation and yielding tota
l turnover numbers for the coenzyme of approximately 1000 for a single roun
d of D-fructose conversion. In optimized batchwise reduction of D-fructose,
a D-mannitol productivity of 2.25 g/(Lh) was obtained for a final product
concentration of 72 g/L and a D-fructose conversion of 80%. D-Mannitol was
crystallized From the ultrafiltered product solution in 97% purity and 85%
recovery, thus also allowing reuse of enzymes for repeated batchwise produc
tion of D-mannitol.