Biochemical localization of the alkaline phosphatase of Bacillus licheniformis as a function of culture age.
Biochemical localization of the enzyme as a function of age of cell culture showed the alkaline phosphatase (orthophosphoric monoester phosphohydrolase, EC 3.1.3.1) activity of Bacillus licheniformis MC14 predominantly in the particulate cell fraction in early- and mid-log cells. However, in late-log and stationary cells, increasing amounts of activity were found in the soluble fraction of lysed cells. Upon protoplast formation of these cells, the activity was released into the soluble fraction. No alkaline phosphatase activity was found in either the cytoplasmic fraction or in the cell medium during any phase of cell growth. The soluble fraction released on protoplast formation that contained alkaline phosphatase activity showed immunological cross-reactivity with antibody to the purified heat--salt-solubilized membrane alkaline phosphatase (F. M. Hulett-Cowling and L. L. Campbell, 1971). Theparticulate membrane fraction containing a firmly associated alkaline phosphatase also showed similar cross-reactivity. Further, the effectiveness of nonionic detergents, ionic detergents, bile salts, and various concentrations of magnesium and sodium as solubilizing agents for this membrane-bound alkaline phosphatase was investigated. Hexadecyl pyridinium chloride (0.03 M) and magnesium and sodium salts (above 0.2 M) were effective solubilizing agents. The substrate specificities of the various fractions were determined and compared to the substrate specificities of the purified membrane alkaline phosphatase.
D000469:Alkaline Phosphatase / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D001407:Bacillus / Q000201:enzymology* / Q000254:growth & development; D001647:Bile Acids and Salts; D002462:Cell Membrane / Q000201:enzymology; D003593:Cytoplasm / Q000201:enzymology; D011523:Protoplasts / Q000201:enzymology; D012995:Solubility; D013347:Subcellular Fractions; D013501:Surface-Active Agents
D001647:Bile Acids and Salts; D013501:Surface-Active Agents; D000469:Alkaline Phosphatase