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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{triangledown}

Ryan D. Woodyer,1* Nathan J. Wymer,1,{dagger} 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.

* Corresponding author. Mailing address: zuChem, Rm. B107, 801 W. Main St., Peoria, IL 61606. Phone: (309) 495-7318. Fax: (309) 495-7319. E-mail: rwoodyer{at}zuchem.com

{triangledown} Published ahead of print on 14 March 2008.

{dagger} Present address: Pfizer, Inc., Groton, CT.

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.



This article has been cited by other articles:

  • Yeom, S.-J., Kim, N.-H., Park, C.-S., Oh, D.-K. (2009). L-Ribose Production from L-Arabinose by Using Purified L-Arabinose Isomerase and Mannose-6-Phosphate Isomerase from Geobacillus thermodenitrificans. Appl. Environ. Microbiol. 75: 6941-6943 [Abstract] [Full Text]  
  • Yeom, S.-J., Ji, J.-H., Kim, N.-H., Park, C.-S., Oh, D.-K. (2009). Substrate Specificity of a Mannose-6-Phosphate Isomerase from Bacillus subtilis and Its Application in the Production of L-Ribose. Appl. Environ. Microbiol. 75: 4705-4710 [Abstract] [Full Text]  


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