Purification and properties of a glycoprotein acid phosphatase from the yeast form of Yarrowia lipolytica*
Paper presented at the XIIth International Spezialized Symposium on Yeast: “Genetics of non‐conventional yeasts”, held in Weimar, GDR, September 13–19, 1987
Abstract
An acid phosphatase from derepressed cells of the yeast form of Yarrowia lipolytica was purified 176‐fold by ammonium sulfate precipitation, chromatography on DEAE‐Sephacel and gel filtration in Biogel A 5‐M. The enzyme is a glycoprotein with a sugar content of 60%. The MICHAELIS constant of the enzyme is 5.5 × 10−4 M for p‐nitrophenyl phosphate as substrate; the isoelectric point estimated by electrofocusing is around 4.6 and the optimum pH is 6.2. Phosphatase activity was destroyed by exposure of the enzyme to 40 °C for 30 min or at pH 3 for 30 min. The purified enzyme shows size heterogeneity within a linear concentration gradient of polyacrylamide (4–20%) which indicates an apparent molecular weight in the range of 90,000–200,000 in the presence of sodium dodecyl sulfate. The heterogeneity of the enzyme is due to its carbohydrate content, as can be demonstrated by gel filtration and by treatment with endoglycosidase H. The carbohydrate‐depleted protein has a molecular weight of 60,000. Aminoacid analysis revealed a high content of aspartic acid, serine, threonine, glycine and alanine. Antibodies against the protein moiety show cross reactivity with the acid phosphatase of Saccharomyces cerevisiae.
Number of times cited: 14
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