Abstract
We overexpressed and purified enzymes involved in the pyrimidine catabolic pathway in the yeast Saccharomyces (Lachancea) kluyveri. A new vector was therefore designed, providing the first specific expression system in Saccharomyces kluyveri. The URC1 gene was overexpressed and a soluble protein obtained and successfully purified using the C-terminally added His-tag. Our system will be used for further studies of the structure and function of the enzymes belonging to the URC pyrimidine degradation pathway.
Acknowledgments
The authors thank the Swedish Research Council (VR) and the Lawski Foundation for their funding. We also thank Cecilia Emanuelsson (Lund University) who kindly helped with the mass spectrometry analysis.