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Article
Zymography of Monophenolase and o-Diphenolase Activities of Polyphenol Oxidase
Analytical Biochemistry (2002)
  • Mamoudou H. DICKO, UniversitĂ© de Ouagadougou
  • Riet HILHORST
  • Harry GRUPPEN
  • Colja C; LAANE
  • Willem J. H. van BERKEL
  • Alphons G. J. VORAGEN
Abstract

A new procedure for the zymography of monophenolase and o-diphenolase activities of polyphenol oxidase (PPO), and peroxidase (POX) is proposed using a highly sensitive, chromogenic nucleophile 3-methyl-2-benzothiazolinone hydrazone (MBTH), which traps quinones. The procedure allowed the distinction between PPO isoenzymes from sorghum and mushroom in the same gel, as well as between monophenolase and o-diphenolase activities of PPO isoenzymes from crude extracts of mushroom. Three isoforms were detected with monophenolase activity, and at least seven isoforms were detected with o diphenolase activity. The procedure also allows the identification of PPO isoforms exhibiting monophenolase activity from a crude extract. The sensitivity, speed and ability to discriminate between mono and o-diphenolase activities could make the newly developed procedure a universal and powerful method for the routine zymography of PPOs and POXs in biological materials. The assay also discriminates the activities of PPOs, POXs and laccases.

Keywords
  • polyphenol oxidase,
  • peroxidase,
  • zymography,
  • activity staining,
  • isoenzymes,
  • mushroom,
  • sorghum
Publication Date
June, 2002
Publisher Statement
Copyright: Academic Press, USA
Citation Information
Mamoudou H. DICKO, Riet HILHORST, Harry GRUPPEN, Colja C; LAANE, et al.. "Zymography of Monophenolase and o-Diphenolase Activities of Polyphenol Oxidase" Analytical Biochemistry Vol. 306 (2002)
Available at: http://works.bepress.com/dicko/4/