Effect of Cu2+, Mn2+ and aromatic compounds on the production of laccase isoforms by Coprinus comatus

Six different extracellular laccase isoforms were identified in submerged cultures of the commercially important edible mushroom, Coprinus comatus. Although laccase activity (similar to 55 IU/L) was readily detectable in unsupplemented control cultures containing 1.6 mu M Cu2+ after 22-day incubation, mean enzyme levels (similar to 150-185 IU/L) were 2.7-3.4-fold higher in cultures supplemented with 0.5-3.0 mM Cu2+. Laccase production was also stimulated by Mn supplementation over the range 0.05-0.8 mM Mn2+, and the peak value of similar to 225 IU/L recorded after 22 days in cultures containing 0.8 mM added Mn2+ was 4.5-fold higher compared with unsupplemented controls. Of 12 aromatic compounds tested for their effect on laccase isozyme production by C. comatus, highest laccase levels (similar to 188 IU/L), equivalent to a 4.4-fold increase compared with unsupplemented controls (similar to 43 IU/L), were recorded after 22 days in cultures supplemented with 3.0 mM caffeic acid. Other aromatic compounds tested all stimulated laccase production, with peak enzyme levels 1.3-3.3-fold higher compared with unsupplemented controls. Extracellular laccase levels in cultures supplemented with optimal concentrations of Mn2+ and caffeic acid together were 38% and 15% lower, respectively, compared with cultures containing the separate supplements. Lac1 was the most abundant laccase isoform produced under all the conditions tested, but marked differences were observed in the production patterns of Lac2-Lac6.