Glycosidases in Carrot Cells in Suspension Culture : Localization and Activity Change during Growth

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Localization of four glycosidases, α-galactosidase (α-Gal), p-galactosidase (β-Gal), α-glucosidase (α-Glu) and p-glucosidase (β-Glu) in suspension-cultured carrot cells was studied. Wall-bound enzymes were made soluble when the cells were converted to protoplasts by cellulase and pectinase. α-Gal was separated into two forms, designated I and II, by chromatography on a Sephadex G-200 colunm. α-Gal I was located exclusively in the cytoplasm whereas α-Gal 11 was found in both the cytoplasmic and cell-wall fractions. The pH optimum was in the neutral region for α-Gal I and in the acidic region for the other glycosidases, including α-Gal II. Both intact cells and protoplasts in suspension culture secreted these glycosidases, except α-Gal I, into the medium. Specific activities of the glycosidases, especially the activity of p-Gal, decreased in the early logarithmic growth phase and increased as cells went through late logarithmic and stationary phases. In protoplast culture, glycosidase activity gradually increased as cell wall regeneration proceeded.
日本植物生理学会の論文