Construction of a novel bioanode for amino acid powered fuel cells through an artificial enzyme cascade pathway
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The construction of a novel bioanode based on l-proline oxidation using a cascade reaction pathway comprised of thermostable dehydrogenases.
A novel multi-enzymatic cascade pathway, containing four kinds of dehydrogenases from thermophiles (dye-linked l-proline dehydrogenase, nicotinamide adenine dinucleotide (NAD)-dependent Δ1-pyrroline-5-carboxylate dehydrogenase, NAD-dependent l-glutamate dehydrogenase and dye-linked NADH dehydrogenase), was designed for the generation of six-electrons from one molecule of l-proline. The current density of the four-dehydrogenase-immobilized electrode, with a voltage of + 450 mV (relative to that of Ag/AgCl), was 226.8 μA/cm2 in the presence of 10 mM l-proline and 0.5 mM ferrocene carboxylate at 50 °C. This value was 4.2-fold higher than that of a similar electrode containing a single dehydrogenase. In addition, about 54% of the initial current in the multi-enzyme cascade bioanode was maintained even after 15 days.
Efficient deep oxidation of l-proline by multiple-enzyme cascade reactions was achieved in our designed electrode. The multi-enzyme cascade bioanode, which was built using thermophilic dehydrogenases, showed high durability at room temperature. The long-term stability of the bioanode indicates that it shows great potential for applications as a long-lived enzymatic fuel cell.
KeywordsEnzymatic fuel cell Thermophilic enzyme Enzyme cascade reaction Dye-linked dehydrogenase
We thank Ms. Yumi Sugamura for technical assistance. We would like to thank Editage (www.editage.jp) for English language editing.
Supplementary Figure 1—SDS-PAGE of purified enzymes making up the l-proline multi-step oxidation pathway. A, Ap-LPDH; B, ThP5CDH; C, GkNADHDH; D, Pi-GDH; M, protein marker; E, purified enzyme.
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Conflict of interest
The authors declare that they have no competing interests.
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