Abstract
Objectives
To investigate the efficiency of a cofactor regeneration enzyme co-expressed with a glycerol dehydrogenase for the production of 1,3-dihydroxyacetone (DHA).
Results
In vitro biotransformation of glycerol was achieved with the cell-free extracts containing recombinant GlyDH (glycerol dehydrogenase from Escherichia coli), LDH (lactate dehydrogenase form Bacillus subtilis) or LpNox1 (NADH oxidase from Lactobacillus pentosus), giving DHA at 1.3 g l−1 (GlyDH/LDH) and 2.2 g l−1 (GlyDH/LpNox1) with total turnover number (TTN) of NAD+ recycling of 6039 and 11100, respectively. Whole cells of E. coli (GlyDH–LpNox1) co-expressing both GlyDH and LpNox1 were constructed and converted 10 g glycerol l−1 to DHA at 0.2–0.5 g l−1 in the presence of zero to 2 mM exogenous NAD+. The cell free extract of E. coli (GlyDH–LpNox) converted glycerol (2–50 g l−1) to DHA from 0.5 to 4.0 g l−1 (8–25 % conversion) without exogenous NAD+.
Conclusions
The disadvantage of the expensive consumption of NAD+ for the production of DHA has been overcome.
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Acknowledgements
This work was financially supported by the Qualified Personnel Foundation of Taiyuan University of Technology (Grant No. tyut-rc201484a) and the Youth Foundation of Taiyuan University of Technology (Grant No. 1205-0020202) and the Scientific and Technological Innovation Programs of Higher Education Institutions in Shanxi (STIP) (Grant No. 2015132).
Supporting information
Supplementary Table 1—The recombinant plasmids and E. coli strains used in this study.
Supplementary Table 2—The primers used for PCR amplification of DNA sequences.
Supplementary Fig. 1—SDS-PAGE analysis of cell free extract of E. coli (LDH), E. coli (LpNox1) and E. coli (GlyDH).
Supplementary Fig. 2—SDS-PAGE analysis of the cell free extract of E. coli (GlyDH–LpNox1) after induction.
Supplementary Fig. 3—Effect of IPTG concentration (a) and induction temperature (b) on GlyDH and LpNox1 expression in E. coli (GlyDH–LpNox1).
Supplementary Fig. 4—Time profiles of cell density (g cdw l−1) and the activity of GlyDH and LpNox1 during fermentation process.
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Zhang, J., Cui, Z., Chang, H. et al. Conversion of glycerol to 1,3-dihydroxyacetone by glycerol dehydrogenase co-expressed with an NADH oxidase for cofactor regeneration. Biotechnol Lett 38, 1559–1564 (2016). https://doi.org/10.1007/s10529-016-2130-3
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DOI: https://doi.org/10.1007/s10529-016-2130-3