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<p>Biorefinery is an innovative concept for resource utilization. To further increase its efficiency, the integration of different biomass conversion technologies (integrated biorefinery) is essential. Biodiesel, a traditional biofuel, generates glycerol as an inevitable byproduct; making it an attractive carbon source for biorefineries due to its abundance, low price, and high degree of reduction. In this work, we propose an integrated two-step process of dark fermentation with <i>Escherichia coli</i> to produce L-malate from glycerol and photofermentation by <i>Rhodobacter capsulatus</i> to convert this L-malate into hydrogen, a clean and sustainable energy carrier. To this end, we optimized an <i>E. coli</i> L-malate-producing strain by overexpressing GlpK in the M4-Δ<i>iclr</i>/pBAD-<i>pck</i> strain and scale up the process from shake flask to bioreactor using waste crude glycerol from a biodiesel biorefinery instead of pure glycerol. Under the optimized conditions, <i>E. coli</i> produced up to 11&#xa0;g/L L-malate in 24&#xa0;h. The culture medium from this dark fermentation was used to formulate an L-malate enriched medium for <i>Rhodobacter</i> that led to the production of 58.0 ± 6 mM H<sub>2</sub> in 90&#xa0;h.</p> Graphical abstract <p></p>

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Production of hydrogen from crude glycerol via an integrated process of L-malate biosynthesis by Escherichia coli and photofermentation by Rhodobacter capsulatus

  • 期刊论文

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  • 发表日期：

    2025年11月10日

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  • Jose L. Marcos,
  • Gema Cabrera,
  • Daniel F. Hernandez,
  • Beatriz Luque,
  • Antonio Valle,
  • Jorge Bolivar

摘要

Biorefinery is an innovative concept for resource utilization. To further increase its efficiency, the integration of different biomass conversion technologies (integrated biorefinery) is essential. Biodiesel, a traditional biofuel, generates glycerol as an inevitable byproduct; making it an attractive carbon source for biorefineries due to its abundance, low price, and high degree of reduction. In this work, we propose an integrated two-step process of dark fermentation with Escherichia coli to produce L-malate from glycerol and photofermentation by Rhodobacter capsulatus to convert this L-malate into hydrogen, a clean and sustainable energy carrier. To this end, we optimized an E. coli L-malate-producing strain by overexpressing GlpK in the M4-Δiclr/pBAD-pck strain and scale up the process from shake flask to bioreactor using waste crude glycerol from a biodiesel biorefinery instead of pure glycerol. Under the optimized conditions, E. coli produced up to 11 g/L L-malate in 24 h. The culture medium from this dark fermentation was used to formulate an L-malate enriched medium for Rhodobacter that led to the production of 58.0 ± 6 mM H2 in 90 h.

Graphical abstract