Planta

, Volume 196, Issue 1, pp 141–147

Separation and characterization of the isoenzymes of wall-bound peroxidase from cultured Zinnia cells during tracheary element differentiation

Authors

  • Yasushi Sato
    • Biological Institute, Faculty of ScienceTohoku University
  • Munetaka Sugiyama
    • Biological Institute, Faculty of ScienceTohoku University
  • Atsushi Komamine
    • Department of Chemical and Biological ScienceJapan Women's University
  • Hiroo Fukuda
    • Biological Institute, Faculty of ScienceTohoku University
Article

DOI: 10.1007/BF00193227

Cite this article as:
Sato, Y., Sugiyama, M., Komamine, A. et al. Planta (1995) 196: 141. doi:10.1007/BF00193227

Abstract

Cell wall-bound peroxidase (EC 1.11.1.7) isoenzymes (P1-P5) from cells of Zinnia elegans L. that were differentiating into tracheary elements were separated and characterized to obtain information about the relationships between these isoenzymes and the biosynthesis of lignin. Fractionation of Zinnia cells by centrifugation in solutions of Percoll revealed that P1, P2, and P5 were present in differentiated tracheary elements. These peroxidase isoenzymes were separated by several column-chromatographic steps. During hydrophobic chromatography on Phenyl Superose, P5 activity was separated into activities P5A and P5B. Enzymatically pure preparations of P1, P3, P5A, and P5B were finally obtained and used for the characterization of each isoenzyme. The optimum pH was 5.5–6.0 for P1, 5.0–7.5 for P3, 5.0 for P5A, and 4.0 for P5B. Each of the isoenzymes oxidized coniferyl alcohol efficiently, whereas p-coumaryl alcohol and sinapyl alcohol were poor substrates for all the isoenzymes. An absolute requirement for Ca2+ ions was demonstrated for P3. Based on these results, possible roles of peroxidase isoenzymes in the formation of lignin during the differentiation of tracheary elements are discussed.

Key words

Cell wallConiferyl alcoholLignificationPeroxidase isoenzymesTracheary element differentiationZinnia

Abbreviations

DAB

diaminobenzidine

GTA

equal proportions of 3,3-dimethylglutaric acid, tris(hydroxymethyl)aminomethane, and 2-amino-2-methyl-1,3-propanediol

TE

tracheary element

Copyright information

© Springer-Verlag 1995