Cloning of orotidine-5′-phosphate decarboxylase (URA3) gene from sourdough yeast Candida milleri CBS 8195

Abstract

The nucleotide sequence of the URA3 gene encoding orotidine-5′-phosphate decarboxylase (OMPDCase) in sourdough yeast Candida milleri CBS 8195 was determined by degenerate PCR and genome walking. Sequence analysis revealed the presence of an openreading frame of 810 bp, encoding 269 amino acid residue protein with the highest identity to the OMPDCase of the yeast Saccharomyces cerevisiae. Phylogenetic analysis of deduced amino acid sequence revealed that it shares a high degree of identity with other yeast OMPDCases. The cloned URA3 gene successfully complemented the ura3 mutation in S. cerevisiae, indicating that it encodes a functional OMPDCase in C. millieri CBS 8195.