Fig. 4

The effect of copper in the secretion of the small laccases. Secretion of a) AmLac, b) ScLac, and c) SvLac in the absence of additional copper supplementation. Spores were inoculated into growth media containing 0 µM of CuSO₄. Samples were taken at defined time points and the cells were separated by centrifugation. The supernatants were aliquoted into two separate samples, one was saved as a control. The second aliquot was reconstituted by supplementing with 100 µM of CuSO₄, incubated overnight and measured on the next day. The reconstituted samples were named medium (R) and cells (R). The reconstituted activities of AmLac, ScLac and SvLac were detected in the medium indicating that secretion was happening independently of copper. The activity results were the average of three biological replicates in three technical replicates each. The comparison of d) AmLac, e) ScLac, or f) SvLac volumetric activities in the absence and presence of copper. The volumetric activities were normalized to WCW. Higher activity was measured in the medium when copper was present at the beginning of the expression