Purification to homogeneity of Candida albicans glucosamine-6-phosphate synthase overexpressed in Escherichia coli

The Candida albicans GFA1 gene encoding glucosamine-6-phosphate synthase, a n enzyme of cell wall biosynthesis pathway in fungi and bacteria, recently an object of interest as a target for the chemotherapy of systemic mycoses, was PCR amplified and cloned to an Escherichia coli expression vector pET2 3b. The activity of the enzyme in the lysates from the overproducing E. col i strain was approximately 50-100 times higher than in the lysates from the control E. coli strain. This abundant overproduction allows to purify mill igram amounts of the enzyme to homogeneity. (C) 2000 Academic Press.