Expression and characterization of Phr2 protein of Candida albicans

Transglycosylases have an import function in yeast the cell wall. These enzymes cleave a molecule of donor, which is followed by transferring a newly created reducing end to a non-reducing end of a molecule of an acceptor. This process enables a growth, expansion and shaping of the yeast cell walls during their life. The aim of this Bachelor Thesis is to characterise the role of Phr2 recombinant protein of Candida albicans in a transglycosylase activity. Using fluorescent paper method, optimal pH and optimal temperature of transglycosylase activity of Phr2 protein were found. The same method was used to establish the acceptor and donor specificity of Phr2p was performed as another method for substrate specificity measurement. High donor and acceptor specificity of Phr2 protein to -1,3-glucan was confirmed. HPLC products of transglycosylase reaction catalysed by Phr2p with laminarin as a donor and L5-SR as an acceptor containing a fluorescent marker were detected by TLC and MALDI-TOF