Purification and biochemical characterization of a new alkali-stable laccase from Trametes sp. isolated in Tunisia: role of the enzyme in olive mill waste water treatment

World J Microbiol Biotechnol. 2013 Nov;29(11):2145-55. doi: 10.1007/s11274-013-1380-7. Epub 2013 May 28.

Abstract

A white-rot basidiomycete, isolated from decayed acacia wood (from Northwest of Tunisia) and identified as Trametes sp, was selected in a broad plate screening because of its ability to decolorize and dephenolize olive oil mill wastewater (OMW) efficiently. The major laccase was purified and characterized as a monomeric protein with apparent molecular mass of 61 kDa (SDS-PAGE). It exhibits high enzyme activity over broad pH and temperature ranges with optimum activity at pH 4.0 and a temperature of 60 °C. The purified laccase is stable at alkaline pH values. The enzyme retained 50 % of its activity after 90 min of incubation at 55 °C. Using ABTS, this laccase presented K m and V max values of 0.05 mM and 212.73 μmoL min(-1) mg(-1), respectively. It has shown a degrading activity towards a variety of phenolic compounds. The purified laccase was partially inhibited by Fe(2+), Zn(2+), Cd(2+) and Mn(2+), while Cu(2+) acted as inducer. EDTA (10 mM) and NaN3 (10 mM) were found to completely inhibit its activity. 73 % OMW was dephenolized after 315 min incubation at 30 °C with 2 U mL(-1) of laccase and 2 mM HBT.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Basidiomycota / classification
  • Biodegradation, Environmental
  • Enzyme Stability
  • Fungal Proteins / genetics
  • Fungal Proteins / isolation & purification
  • Fungal Proteins / metabolism
  • Kinetics
  • Laccase / genetics
  • Laccase / isolation & purification*
  • Laccase / metabolism*
  • Olea / metabolism*
  • Polyphenols / metabolism
  • Substrate Specificity
  • Temperature
  • Trametes / enzymology*
  • Trametes / genetics
  • Trametes / isolation & purification*
  • Tunisia
  • Wastewater / chemistry*
  • Water Purification
  • Wood / microbiology

Substances

  • Fungal Proteins
  • Polyphenols
  • Waste Water
  • Laccase