The Protein Journal

, Volume 36, Issue 2, pp 98–107 | Cite as

Complete Amino Acid Sequence of a Copper/Zinc-Superoxide Dismutase from Ginger Rhizome

  • Yuki Nishiyama
  • Tamo Fukamizo
  • Kazunari Yoneda
  • Tomohiro Araki
Article

Abstract

Superoxide dismutase (SOD) is an antioxidant enzyme protecting cells from oxidative stress. Ginger (Zingiber officinale) is known for its antioxidant properties, however, there are no data on SODs from ginger rhizomes. In this study, we purified SOD from the rhizome of Z. officinale (Zo-SOD) and determined its complete amino acid sequence using N terminal sequencing, amino acid analysis, and de novo sequencing by tandem mass spectrometry. Zo-SOD consists of 151 amino acids with two signature Cu/Zn-SOD motifs and has high similarity to other plant Cu/Zn-SODs. Multiple sequence alignment showed that Cu/Zn-binding residues and cysteines forming a disulfide bond, which are highly conserved in Cu/Zn-SODs, are also present in Zo-SOD. Phylogenetic analysis revealed that plant Cu/Zn-SODs clustered into distinct chloroplastic, cytoplasmic, and intermediate groups. Among them, only chloroplastic enzymes carried amino acid substitutions in the region functionally important for enzymatic activity, suggesting that chloroplastic SODs may have a function distinct from those of SODs localized in other subcellular compartments. The nucleotide sequence of the Zo-SOD coding region was obtained by reverse-translation, and the gene was synthesized, cloned, and expressed. The recombinant Zo-SOD demonstrated pH stability in the range of 5–10, which is similar to other reported Cu/Zn-SODs, and thermal stability in the range of 10–60 °C, which is higher than that for most plant Cu/Zn-SODs but lower compared to the enzyme from a Z. officinale relative Curcuma aromatica.

Keywords

Zingiber officinale Ginger rhizome Cu/Zn-superoxide dismutase Amino acid sequencing 

Abbreviations

Cu/Zn-SOD

Copper/zinc-superoxide dismutase

Zo-SOD

SOD from Zingiber officinale rhizome

ROS

Reactive oxygen species

\(\text{O}_{2}^{-\cdot}\)

Superoxide radical

SDS–PAGE

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

MALDI/TOF

Matrix-assisted laser desorption/ionization/time of flight

ESI Q/TOF

Electrospray ionization/hybrid quadrupole time of flight

MS/MS

Tandem mass spectrometry

Notes

Acknowledgements

This study was supported in part by the Tokai University Educational System. The manuscript has been edited by a professional English-speaking editor (Editage; http://www.editage.jp). The authors declare that we have no conflicts of interest. This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Yuki Nishiyama
    • 1
  • Tamo Fukamizo
    • 2
  • Kazunari Yoneda
    • 1
  • Tomohiro Araki
    • 1
  1. 1.Department of Bioscience, School of AgricultureTokai UniversityKumamotoJapan
  2. 2.Department of Advanced BioscienceKinki UniversityNaraJapan

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