<p>1,12-Dodecanediol is a high-value chemical widely used in the polymer and fine chemical industries. However, its industrial production is currently based on petroleum-derived processes associated with high energy consumption and environmental concerns. To address these limitations, microbial bioconversion of 1,12-dodecanedioic acid (C12 diacid) to 1,12-dodecanediol (C12 diol) has been investigated as a sustainable production route. In this pathway, carboxylic acid reductase (CAR) requires ATP and NADPH as essential cofactors, making efficient cofactor supply an important factor for achieving high conversion efficiency. In this study, a recombinant <i>Escherichia coli</i>–based biocatalytic system was developed to enhance C12 diol production through ATP regeneration. ATP availability was identified as an important factor for whole-cell bioconversion, and a polyphosphate kinase 2 (PPK2) from <i>Erysipelotrichaceae bacterium</i> was introduced to regenerate ATP from AMP, increasing the conversion yield from 22.0% to 31.8% at 40 mM substrate. Supplementation with sodium hexametaphosphate (SHMP, polyP<sub>6</sub>) further improved diol production, reaching 19.5 mM C12 diol with a conversion yield of 48.8%. In the cell-lysate-based conversion, NADPH and NADH exhibited comparable conversion efficiencies up to 70 mM MgCl<sub>2</sub>. Diol production was influenced by MgCl<sub>2</sub> concentration, with the highest conversion obtained at 100 mM MgCl<sub>2</sub> in the presence of NADPH, yielding approximately 28 mM C12 diol (~ 70% conversion) within 1&#xa0;h. Overall, this study highlights the importance of ATP regeneration and cofactor supply for improving C12 diol production.</p> Graphical abstract <p></p>

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ATP regeneration-driven biocatalytic production of 1,12-dodecanediol with improved cofactor supply

  • Yoon Jung Jung,
  • Gaeul Kim,
  • Kyungjae Yu,
  • Byung Wook Lee,
  • Jung Bin Shin,
  • Jung-Oh Ahn,
  • Si Jae Park,
  • See-Hyoung Park,
  • Hyun Gi Koh,
  • Kyungmoon Park

摘要

1,12-Dodecanediol is a high-value chemical widely used in the polymer and fine chemical industries. However, its industrial production is currently based on petroleum-derived processes associated with high energy consumption and environmental concerns. To address these limitations, microbial bioconversion of 1,12-dodecanedioic acid (C12 diacid) to 1,12-dodecanediol (C12 diol) has been investigated as a sustainable production route. In this pathway, carboxylic acid reductase (CAR) requires ATP and NADPH as essential cofactors, making efficient cofactor supply an important factor for achieving high conversion efficiency. In this study, a recombinant Escherichia coli–based biocatalytic system was developed to enhance C12 diol production through ATP regeneration. ATP availability was identified as an important factor for whole-cell bioconversion, and a polyphosphate kinase 2 (PPK2) from Erysipelotrichaceae bacterium was introduced to regenerate ATP from AMP, increasing the conversion yield from 22.0% to 31.8% at 40 mM substrate. Supplementation with sodium hexametaphosphate (SHMP, polyP6) further improved diol production, reaching 19.5 mM C12 diol with a conversion yield of 48.8%. In the cell-lysate-based conversion, NADPH and NADH exhibited comparable conversion efficiencies up to 70 mM MgCl2. Diol production was influenced by MgCl2 concentration, with the highest conversion obtained at 100 mM MgCl2 in the presence of NADPH, yielding approximately 28 mM C12 diol (~ 70% conversion) within 1 h. Overall, this study highlights the importance of ATP regeneration and cofactor supply for improving C12 diol production.

Graphical abstract