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Biochemistry (Moscow)

, Volume 74, Issue 9, pp 1027–1034 | Cite as

Purification and characteristics of an enzyme with both bilirubin oxidase and laccase activities from mycelium of the basidiomycete Pleurotus ostreatus

  • Y. G. PakhadniaEmail author
  • N. I. Malinouski
  • A. G. Lapko
Article

Abstract

A homogenous enzyme with both bilirubin oxidase and laccase activities was isolated from a submerged culture of the basidiomycete Pleurotus ostreatus mycelium and characterized. The yield of the enzyme was 127 μg/g dry biomass of the mycelium. The specific activity of the enzyme was 21 and 261 U/mg to bilirubin and to a laccase substrate ABTS, respectively. The intracellular phenol oxidase from the P. ostreatus mycelium was identified as bilirubin oxidase with the amino acid sequence highly homologous to that of the pox2 gene-encoded product. The enzyme displayed the maximal laccase activity at 50–55°C to all substrates examined, whereas the pH optimum was substrate-dependent and changed from 3.0 for ABTS to 7.0 for syringaldazine and guaiacol. The enzyme maintained catalytic activity within a broad pH range but was inactivated at pH 4.0. The enzyme was thermostable but very sensitive to metal chelating inhibitors. Trypan Blue (5 mg/liter) was completely decolorizated upon 3 h of incubation with the bilirubin oxidase (20 mU/ml) at room temperature.

Key words

mycelium Pleurotus ostreatus oxidoreductase bilirubin oxidase laccase 

Abbreviations

ABTS

2,2′-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid)

BOX

bilirubin oxidase

DMP

2,6-dimethoxyphenol

DTT

dithiothreitol

SGZ

syringaldazine

TB

Trypan Blue

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Copyright information

© Pleiades Publishing, Ltd. 2009

Authors and Affiliations

  • Y. G. Pakhadnia
    • 1
    Email author
  • N. I. Malinouski
    • 2
  • A. G. Lapko
    • 1
  1. 1.Sakharov International Environmental UniversityMinskBelarus
  2. 2.Department of BiochemistryUniversity of NebraskaLincolnUSA

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