Purification of Protein Disulfide Isomerase from a Thermophilic Fungus

Abstract

A protein disulfide isomerase (PDI) was purified to homogeneity from the thermophilic fungus Humicola insolens by a rapid three-step procedure, anion-exchange chromatography, concanavalin A-affinity chromatography, and reverse phase high performance liquid chromatography. Forty-oneμg of PDI was obtained from 100g of wet mycelium. Concanavalin A-Sepharose chromatography is available for purification of the fungal PDI, indicating that the enzyme is also glycosylated like the yeast PDI. The fungal PDI exists as a dimer (2 x 60 kDa), has a pI of 3.5, and is fairly heat-stable. The amino acid composition of the PDI is similar to those of yeast and bovine liver PDI, and the high content of acidic amino acid residues agrees with the lower acidic pI.