PURIFICATION AND CHARACTERIZATION OF AN INVERTASE FROM CANDIDA-UTILIS- COMPARISON WITH NATURAL AND RECOMBINANT YEAST INVERTASES

A periplasmic invertase from the yeast Candida utilis was purified to homogeneity from cells fully derepressed for invertase synthesis. The enzyme was purified by successive Sephacryl S-300, and affinity chroma tography and shown to be a dimeric glycoprotein composed of two identi cal monomer subunits with an apparent molecular mass of 150 kDa. After EndoH treatment, the deglycosylated protein showed an apparent molecu lar weight of 60 kDa. The apparent K-m values for sucrose and raffinos e were 11 and 150 mM, respectively, similar to those reported in Sacch aromyces cerevisiae. The range of optimum temperature was 60-75 degree s C. The optimum pH was 5.5 and the enzyme was stable over pH range 3- 6. (C) 1997 Elsevier Science B.V.