The dye (595 nm) was decolorized by 80% within 24 h and almost completely decolorized within 48 h after addition. The nitrogen in the reactor was exhausted in 7 days and the glucose exhausted in 12 days whereupon more glucose was added to the reactor (Lonergan et al. 1995b). There was a small increase in nitrogen towards the end of the cycle due to autolysis.
Laccase activity was detected on day 2, peaked at a very large 60 U/ml on day 8 and fell to a minimum on day 12. It then rose again towards the end of the cycle as the additional glucose was added.
In this series of experiments laccases were purified from 100 ml samples removed during the reactor operation on day 3, D3 + 30, 4, 20, 23, and 25 by a slight modification to the method reported (Schliephake et al. 2000). The elution profiles for all samples were similar to those of the laccase purification carried out on the Mono Q anion exchanger. The elution pattern for days 4 and 23 are shown in Figure 4. The anomaly in enzyme elution times (day 4,18.5 min and day 23, 15.5 min) was examined.
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