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Enhancement of NAD(H) pool for formation of oxidized biochemicals in Escherichia coli

  • Fermentation, Cell Culture and Bioengineering - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

The NAD+/NADH ratio and the total NAD(H) play important roles for whole-cell biochemical redox transformations. After the carbon source is exhausted, the degradation of NAD(H) could contribute to a decline in the rate of a desired conversion. In this study, methods to slow the native rate of NAD(H) degradation were examined using whole-cell Escherichia coli with two model oxidative NAD+-dependent biotransformations. A high phosphate concentration (50 mM) was observed to slow NAD(H) degradation. We also constructed E. coli strains with deletions in genes coding several enzymes involved in NAD+ degradation. In shake-flask experiments, the total NAD(H) concentration positively correlated with conversion of xylitol to l-xylulose by xylitol 4-dehydrogenase, and the greatest conversion (80%) was observed using MG1655 nadR nudC mazG/pZE12-xdh/pCS27-nox. Controlled 1-L batch processes comparing E. coli nadR nudC mazG with a wild-type background strain demonstrated a 30% increase in final l-xylulose concentration (5.6 vs. 7.9 g/L) and a 25% increase in conversion (0.53 vs. 0.66 g/g). MG1655 nadR nudC mazG was also examined for the conversion of galactitol to l-tagatose by galactitol 2-dehydrogenase. A batch process using 15 g/L glycerol and 10 g/L galactitol generated over 9.4 g/L l-tagatose, corresponding to 90% conversion and a yield of 0.95 g l-tagatose/g galactitol consumed. The results demonstrate the value of minimizing NAD(H) degradation as a means to improve NAD+-dependent biotransformations.

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Acknowledgements

The authors thank Sarah Lee for the technical assistance.

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Authors and Affiliations

  1. School of Chemical Materials and Biomedical Engineering, University of Georgia, Athens, GA, 30602, USA

    Qi Han & Mark A. Eiteman

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  1. Qi Han
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  2. Mark A. Eiteman
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Correspondence to Mark A. Eiteman.

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Han, Q., Eiteman, M.A. Enhancement of NAD(H) pool for formation of oxidized biochemicals in Escherichia coli. J Ind Microbiol Biotechnol 45, 939–950 (2018). https://doi.org/10.1007/s10295-018-2072-y

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  • DOI: https://doi.org/10.1007/s10295-018-2072-y

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