Sporulation in Bacillus subtilis 168. Comparison of alkaline phosphatase from sporulating and vegetative cells.
1. The purification of the ;vegetative' alkaline phosphatase of Bacillus subtilis 168 was simplified by ionic elution of the enzyme from intact cells. 2. The enzyme has a molecular weight of about 70000 and treatment of the enzyme with 10mm-hydrochloric acid or 6.0m-guanidine hydrochloride, beta-mercaptoethanol (0.1m) gives rise to enzymically inactive subunits. 3. The amino acid composition of the enzyme was determined. The N-terminal residue determined by the DNS chloride method is glycine. 4. The properties of this enzyme were compared with the ;sporulation' alkaline phosphatase of the same strain. 5. Although the ;sporulation' enzyme differs from the ;vegetative' enzyme in its physiology of appearance and apparent mRNA stability, an examination of properties of the enzymes revealed no differences. 6. The enzyme from both cell forms is bound to the particulate fraction of cell extracts, but can be solubilized by high concentrations of magnesium chloride; removal of the magnesium chloride, by dialysis, results in precipitation of both enzymes. Both enzymes can be removed from intact cells by ionic elution. 7. The ;vegetative' and ;sporulation' enzymes have identical pH optima, K(m) and K(i) values and electrophoretic mobilities in cellulose acetate. 8. Their half-life is 28min at 65 degrees C and their Q(10) is 1.25. 9. The molecular size determined by gel filtration on Sephadex G-100 is about 69000. 10. ;Vegetative' and ;sporulation' forms gave precipitin lines that were continuous and non-spurred when tested against antiserum prepared against the ;vegetative' enzyme. 11. The ;sporulation' alkaline phosphatase appears to be associated with stage II of sporulation and appears to be induced by something specifically concerned in sporulation and not by phosphate starvation.
D000469:Alkaline Phosphatase / Q000032:analysis*; D000596:Amino Acids / Q000032:analysis; D001412:Bacillus subtilis / Q000201:enzymology*; D002850:Chromatography, Gel; D004586:Electrophoresis; D006146:Guanidines; D006851:Hydrochloric Acid; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera; D005779:Immunodiffusion; D008274:Magnesium; D008623:Mercaptoethanol; D008970:Molecular Weight; D012333:RNA, Messenger; D013170:Spores / Q000201:enzymology*
D000596:Amino Acids; D006146:Guanidines; D007106:Immune Sera; D012333:RNA, Messenger; D008623:Mercaptoethanol; D000469:Alkaline Phosphatase; D008274:Magnesium; D006851:Hydrochloric Acid