Skip to main content
SpringerLink
Log in

Construction of a novel bioanode for amino acid powered fuel cells through an artificial enzyme cascade pathway

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Objective

The construction of a novel bioanode based on l-proline oxidation using a cascade reaction pathway comprised of thermostable dehydrogenases.

Results

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.

Conclusions

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Adams MW (1993) Enzymes and proteins from organisms that grow near and above 100 degrees C. Annu Rev Microbiol 47:627–658

    Article  CAS  PubMed  Google Scholar 

  • Akers NL, Moore CM, Minteer SD (2005) Development of alcohol/O2 biofuel cells using salt-extracted tetrabutylammonium bromide/Nafion membranes to immobilize dehydrogenase enzymes. Electrochim Acta 50:2521–2525

    Article  CAS  Google Scholar 

  • Arechederra RL, Treu BL, Minteer SD (2007) Development of glycerol/O2 biofuel cell. J Power Sources 173:156–161

    Article  CAS  Google Scholar 

  • Cosnier S, Le Goff A, Holzinger M (2014) Towards glucose biofuel cells implanted in human body for powering artificial organs. Electrochem Commun 38:19–23

    Article  CAS  Google Scholar 

  • Inagaki E, Takahashi H, Kuroishi C, Tahirov TH (2005) Crystallization and avoiding the problem of hemihedral twinning in crystals of Δ1-pyrroline-5-carboxylate dehydrogenase from Thermus thermophilus. Acta Crystallogr F 61:609–611

    Article  CAS  Google Scholar 

  • Inagaki E, Ohshima N, Takahashi H, Kuroishi C, Yokoyama S, Tahirov TH (2006) Crystal structure of Thermus thermophilus Δ1-pyrroline-5-carboxylate dehydrogenase. J Mol Biol 362:490–501

    Article  CAS  PubMed  Google Scholar 

  • Kujo C, Sakuraba H, Nunoura N, Ohshima T (1999) The NAD-dependent glutamate dehydrogenase from the hyperthermophilic archaeon Pyrobaculum islandicum: cloning, sequencing, and expression of the enzyme gene. Biochim Biophys Acta 1434:365–371

    Article  CAS  PubMed  Google Scholar 

  • Li WF, Zhou XX, Lu P (2005) Structural features of thermozymes. Biotechnol Adv 23:271–281

    Article  CAS  PubMed  Google Scholar 

  • Ohshima T, Soda K (1989) Thermostable amino acid dehydrogenases: application and gene cloning. Trends Biotechnol 7:210–214

    Article  CAS  Google Scholar 

  • Palmore GTR, Bertschy H, Bergens SH, Whitesides GM (1998) A methanol/dioxygen biofuel cell that uses NAD+-dependent dehydrogenases as catalysts: application of an electro-enzymatic method to regenerate nicotinamide adenine dinucleotide at low overpotentials. J Electroanal Chem 443:155–161

    Article  CAS  Google Scholar 

  • Sakamoto H, Uchii T, Yamaguchi K, Koto A, Takamura EI, Satomura T, Sakuraba H, Ohshima T, Suye SI (2015) Construction of a biocathode using the multicopper oxidase from the hyperthermophilic archaeon, Pyrobaculum aerophilum: towards a long-life biobattery. Biotechnol Lett 37:1399–1404

    Article  CAS  PubMed  Google Scholar 

  • Sakamoto H, Komatsu T, Yamasaki K, Satomura T, Suye SI (2017) Design of a multi-enzyme reaction on an electrode surface for an l-glutamate biofuel anode. Biotechnol Lett 39:235–240

    Article  CAS  PubMed  Google Scholar 

  • Satomura T, Hara Y, Suye S, Sakuraba H, Ohshima T (2012) Gene expression and characterization of a third type of dye-linked l-proline dehydrogenase from the aerobic hyperthermophilic archaeon, Aeropyrum pernix. Biosci Biotechnol Biochem 76:589–593

    Article  CAS  PubMed  Google Scholar 

  • Satomura T, Hayashi J, Sakamoto H, Nunoura T, Takaki Y, Takai K, Takami H, Ohshima T, Sakuraba H, Suye SI (2018a) d-Lactate electrochemical biosensor prepared by immobilization of thermostable dye-linked d-lactate dehydrogenase from Candidatus Caldiarchaeum subterraneum. J Biosci Bioeng 126:425–430

    Article  CAS  PubMed  Google Scholar 

  • Satomura T, Hayashi J, Ohshida T, Sakuraba H, Ohshima T, Suye S (2018b) Enzymological characteristics of a novel archaeal dye-linked d-lactate dehydrogenase showing loose binding of FAD. Extremophiles 22:975–981

    Article  CAS  PubMed  Google Scholar 

  • Tonooka A, Komatsu T, Tanaka S, Sakamoto H, Satomura T, Suye S (2018) A l-proline/O2 biofuel cell using l-proline dehydrogenase (LPDH) from Aeropyrum pernix. Mol Biol Rep 45:1821–1825

    Article  CAS  PubMed  Google Scholar 

  • Xu S, Minteer SD (2011) Enzymatic biofuel cell for oxidation of glucose to CO2. ACS Catal 2:91–94

    Article  CAS  Google Scholar 

  • Zhu Z, Sun F, Zhang X, Zhang YH (2012) Deep oxidation of glucose in enzymatic fuel cells through a synthetic enzymatic pathway containing a cascade of two thermostable dehydrogenases. Biosens Bioelectron 36:110–115

    Article  CAS  PubMed  Google Scholar 

  • Zhu Z, Tam TK, Sun F, You C, Zhang YHP (2014) A high-energy-density sugar biobattery based on a synthetic enzymatic pathway. Nat Commun 5:3026

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Ms. Yumi Sugamura for technical assistance. We would like to thank Editage (www.editage.jp) for English language editing.

Supplementary information

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.

Author information

Authors and Affiliations

  1. Division of Engineering, Faculty of Engineering, University of Fukui, 3-9-1 Bunkyo, Fukui, 910-8507, Japan

    Takenori Satomura, Hiroaki Sakamoto & Shin-ichiro Suye

  2. Organization for Life Science Advancement Programs, University of Fukui, 3-9-1 Bunkyo, Fukui, 910-8507, Japan

    Takenori Satomura & Shin-ichiro Suye

  3. Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, 3-9-1 Bunkyo, Fukui, 910-8507, Japan

    Kousaku Horinaga & Shino Tanaka

  4. Department of Frontier Fiber Technology and Science, Graduate School of Engineering, University of Fukui, 3-9-1 Bunkyo, Fukui, 910-8507, Japan

    Eiichiro Takamura

  5. Department of Applied Biological Science, Faculty of Agriculture, Kagawa University, 2393 Ikenobe, Miki-cho, Kita-gun, Kagawa, 761-0795, Japan

    Haruhiko Sakuraba

  6. Department of Biomedical Engineering, Faculty of Engineering, Osaka Institute of Technology, 5-16-1 Ohmiya, Asahi-ku, Osaka, 535-8585, Japan

    Toshihisa Ohshima

Authors
  1. Takenori Satomura
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kousaku Horinaga
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shino Tanaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Eiichiro Takamura
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hiroaki Sakamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Haruhiko Sakuraba
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Toshihisa Ohshima
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Shin-ichiro Suye
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Takenori Satomura.

Ethics declarations

Conflict of interest

The authors declare that they have no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PPTX 8907 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Satomura, T., Horinaga, K., Tanaka, S. et al. Construction of a novel bioanode for amino acid powered fuel cells through an artificial enzyme cascade pathway. Biotechnol Lett 41, 605–611 (2019). https://doi.org/10.1007/s10529-019-02664-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-019-02664-8

Keywords

Search

Navigation