(509b) Cloning and Expression of Laccase From Trametes Versicolor in Saccharomyces Cerevisiae Using a Novel Vector System

The long-term goal of this research is to increase efficiency and decrease cost of ethanol fermentation of lignocellulosic feedstocks by combining pre-treatment using laccase enzyme and subsequent fermentation to ethanol through simultaneous saccharification and fermentation paradigms. The first step is to develop a genetically engineered yeast strain capable of degrading lignocellulosic feedstocks to accomplish a consolidated bioprocessing operation. In the study, the Trametes versicolor laccase gene from the vector pBARLAC has been amplified, cloned using the Gateway? vector system (pENTR DTOPO) and LR Clonase II enzyme mix, and transformed and expressed in an auxotrophic strain of Saccharomyces cerevisiae PJ69-4 diploid, using a small ubiquitin-related modifier (SUMO) vector system with histidine as the selectable marker (pSUMOduo-HIS). Laccase activity from the newly constructed recombinant yeast strain, YT2-2, has been determined in the presence of different substrates.