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Appl. Environ. Microbiol. doi:10.1128/AEM.02768-07
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Efficient Production of L-Ribose with a Recombinant E. coli Biocatalyst

zuChem, Inc., 2225 W. Harrison., Suite F, Chicago, IL 60612, USA, and Fermentation Biotechnology Research Unit, National Center for Agricultural Utilization Research, Agricultural Research Service, U. S. Department of Agriculture, Peoria, IL 61604, USA

^* To whom correspondence should be addressed. Email: rwoodyer{at}zuchem.com.

	Abstract

A new synthetic platform with potential for the production of several rare sugars, with l-ribose being 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 with their l-sugar counterparts including ribitol to l-ribose. Recombinant MDH expression was successfully achieved in active form 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 this system was improved using shaken flask experiments. It was determined that the addition of 50-500 µM ZnCl₂ and 5 g/L glycerol both improved production. The final achieved conversions were >70% at a concentration of 40 g/L and >50% at a concentration of 100 g/L. The best conditions determined were then scaled up to a 1 L fermentation that resulted in a 55% conversion of 100 g/L ribitol in 72 hours for a volumetric productivity of 17.4 gL^-1d^-1. This system represents a significantly improved method for the large scale production of l-ribose.