, Volume 22, Issue 6, pp 975–981 | Cite as

Enzymological characteristics of a novel archaeal dye-linked d-lactate dehydrogenase showing loose binding of FAD

  • Takenori SatomuraEmail author
  • Junji Hayashi
  • Tatsuya Ohshida
  • Haruhiko Sakuraba
  • Toshihisa Ohshima
  • Shin-ichiro Suye
Original Paper


A gene-encoding a dye-linked d-lactate dehydrogenase (Dye-DLDH) homolog was identified in the genome of the hyperthermophilic archaeon Thermoproteus tenax. The gene was expressed in Escherichia coli and the product was purified to homogeneity. The recombinant protein exhibited highly thermostable Dye-DLDH activity. To date, four types of Dye-DLDH have been identified in hyperthermophilic archaea (in Aeropyrum pernix, Sulfolobus tokodaii, Archaeoglobus fulgidus, and Candidatus Caldiarchaeum subterraneum). The amino acid sequence of T. tenax Dye-DLDH showed the highest similarity (45%) to A. pernix Dye-DLDH, but neither contained a known FAD-binding motif. Nonetheless, both homologs required FAD for enzymatic activity, suggesting that FAD binds loosely to the enzyme and is easily released unlike in other Dye-DLDHs. Our findings indicate that Dye-DLDHs from T. tenax and A. pernix are a novel type of Dye-DLDH characterized by loose binding of FAD.


d-Lactate Dye-linked dehydrogenase FAD Hyperthermophilic archaea Thermostable enzyme 





2,6-DichloroindophenolHEPES, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid


p-Iodonitrotetrazolium violetMTT, 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide



We thank Ms. Manami Oi, Mr. Kazuya Umebayashi, and Ms. Sayuri Yoshihara for the technical assistance. We would like to thank Editage ( for English language editing.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.


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Copyright information

© Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  • Takenori Satomura
    • 1
    • 2
    Email author
  • Junji Hayashi
    • 3
  • Tatsuya Ohshida
    • 4
  • Haruhiko Sakuraba
    • 4
  • Toshihisa Ohshima
    • 5
  • Shin-ichiro Suye
    • 1
    • 2
  1. 1.Division of Engineering, Faculty of EngineeringUniversity of FukuiFukuiJapan
  2. 2.Organization for Life Science Advancement ProgramsUniversity of FukuiFukuiJapan
  3. 3.Department of Biotechnology, College of Life SciencesRitsumeikan UniversityKusatsuJapan
  4. 4.Department of Applied Biological Science, Faculty of AgricultureKagawa UniversityKagawaJapan
  5. 5.Department of Biomedical Engineering, Faculty of EngineeringOsaka Institute of TechnologyOsakaJapan

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