The lcc1 gene encoding a laccase from T. versicolor DSM11269
was expressed for the first time in Y. lipolytica. Using a molecular
evolution strategy combining random and site-directed mutagenesis,
a double variant (L185P/Q214K) was identified. It showed
2.4- and 2.8-fold increase in kcat/Km for ABTS and 2,6-DMP, respectively.
This study thus demonstrates the strong potential of Y.
lipolytica as platform for the engineering of novel biotechnological
relevant laccases. To isolate efficient enzymes for dye decolorization,
further rounds of evolution will be performed, using specific
screening protocols based on dye decolorization in the presence
of an appropriate mediator.