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Applied and Environmental Microbiology, May 2008, p. 2967-2975, Vol. 74, No. 10
0099-2240/08/$08.00+0     doi:10.1128/AEM.02768-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Efficient Production of L-Ribose with a Recombinant Escherichia coli Biocatalyst

Ryan D. Woodyer,^1* Nathan J. Wymer,^1, F. Michael Racine,¹ Shama N. Khan,¹ and Badal C. Saha²

zuChem, Inc., 2225 W. Harrison, Suite F, Chicago, Illinois 60612,¹ Fermentation Biotechnology Research Unit, National Center for Agricultural Utilization Research, Agricultural Research Service, U.S. Department of Agriculture, Peoria, Illinois 61604²

Received 7 December 2007/ Accepted 8 March 2008

A new synthetic platform with potential for the production of several rare sugars, with L-ribose as the model target, is described. The gene encoding the unique NAD-dependent mannitol-1-dehydrogenase (MDH) from Apium graveolens (garden celery) was synthetically constructed for optimal expression in Escherichia coli. This MDH enzyme catalyzes the interconversion of several polyols and their L-sugar counterparts, including the conversion of ribitol to L-ribose. Expression of recombinant MDH in the active form was successfully achieved, and one-step purification was demonstrated. Using the created recombinant E. coli strain as a whole-cell catalyst, the synthetic utility was demonstrated for production of L-ribose, and the system was improved using shaken flask experiments. It was determined that addition of 50 to 500 µM ZnCl₂ and addition of 5 g/liter glycerol both improved production. The final levels of conversion achieved were >70% at a concentration of 40 g/liter and >50% at a concentration of 100 g/liter. The best conditions determined were then scaled up to a 1-liter fermentation that resulted in 55% conversion of 100 g/liter ribitol in 72 h, for a volumetric productivity of 17.4 g liter^–1 day^–1. This system represents a significantly improved method for the large-scale production of L-ribose.

Published ahead of print on 14 March 2008.

Present address: Pfizer, Inc., Groton, CT.