[ORF19.2286 Gene: Isolation and Purification of Deoxyhypusine Hydroxylase from the Human Pathogenic Yeast Candida...

Candida albicans (C. albicans) is a fungal pathogen that causes infections of the wet body surfaces and the blood in immunocompromised patients or individuals with imbalanced microflora. Since the cases of clinically meaningful candidosis are on the rise, efficient С. albicans therapy is in a high demand. Informed drug design requires well-characterized С. albicans targets, including these aimed at disrupting its post-translational modifications. C. albicans ORF19.2286 gene encodes an ortholog of human deoxyhypusine hydroxylase (DOHH). Here, this ORF was cloned from the SC5314 strain and re-expressed in Escherichia coli as sGB1 - CaDOHH construct with 6xHis tag on the N-terminus of the fusion protein, then purified, and GB1-tag was removed with Tobacco etch virus (TEV) protease. Several amino acid sequence differences between C. albicans and animal DOHHs were noted, and are useful for a selection of the binding sites for antimicrobials in CaDOHH. We present the protocol for the heterologous expression and purification of C. albicans DOHH, which is suitable for further crystallization.