Conversion of glycerol to 1,3-dihydroxyacetone by glycerol dehydrogenase co-expressed with an NADH oxidase for cofactor regeneration

Biotechnol Lett. 2016 Sep;38(9):1559-64. doi: 10.1007/s10529-016-2130-3. Epub 2016 May 27.


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.

Keywords: 1,3-Dihydroxyacetone; Cofactor regeneration; Glycerol; Glycerol dehydrogenase; NADH oxidase.

MeSH terms

  • Dihydroxyacetone / metabolism*
  • Glycerol / metabolism*
  • Multienzyme Complexes / metabolism*
  • NADH, NADPH Oxidoreductases / metabolism*
  • Sugar Alcohol Dehydrogenases / metabolism*


  • Multienzyme Complexes
  • Sugar Alcohol Dehydrogenases
  • glycerol dehydrogenase
  • NADH oxidase
  • NADH, NADPH Oxidoreductases
  • Dihydroxyacetone
  • Glycerol