Partial purification and characterization of polyphenoloxidase from culinary-medicinal royal Sun mushroom (the Himematsutake), Agaricus brasiliensis S. Wasser et al. (Agaricomycetideae)

Akiko Matsumoto-Akanuma, Satoshi Akanuma, Masuro Motoi, Akihiko Yamagishi, Naohito Ohno

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The Royal Sun mushroom, the Himematsutake culinary-medicinal mushroom, Agaricus brasiliensis has several polyphenoloxidase activities in a broad sense. Here we report the partial purification of tyrosinase-type polyphenoloxidase (PPO). PPO is purified from A. brasiliensis without browning using a two-phase partitioning with Triton X-114 and ammonium sulfate fractionation. Partially denaturing SDS-PAGE (sodium dodecyl sulfate-polyacrylamide electrophoresis) staining with L-3,4-dihydroxyphenylalanine was performed and the indicated molecular sizes were approximately 70 kDa and 45 kDa. The purified enzyme is in its latent state and can be activated maximally in the presence of 1.6 mM sodium dodecyl sulfate (SDS). This enzyme catalyzes two distinct reactions, monophenolase and diphenolase activity, and the monophenolase activity showed a lag time typical of polyphenoloxidase. The K m value for 4-tert-butylcatechol was quite similar in the presence and absence of SDS, but the apparent V max value was increased 2.0-fold by SDS. Mimosine was a typical competitive inhibitor with K i values of 138.2 μM and 281.0 μM n the presence and absence of SDS, respectively.

Original languageEnglish
Pages (from-to)73-82
Number of pages10
JournalInternational Journal of Medicinal Mushrooms
Volume13
Issue number1
DOIs
Publication statusPublished - 2011
Externally publishedYes

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Keywords

  • Agaricus brasiliensis
  • Medicinal mushrooms
  • Polyphenoloxidase
  • Royal Sun mushroom
  • SDS activation
  • Triton X-114
  • Tyrosinase

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Drug Discovery
  • Pharmacology

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