Biochemical Studies of Bacterial Sporulation IV. Inorganic Pyrophosphatase of Vegetative Cells and Spores of Bacillus megaterium

Abstract

Inorganic pyrophosphatase activities extracted from vegetative cells and spores ol Bacillius Imnegateriulmz were compared and found to be similar in behavior on polyacrylamide gel electrophoresis, in metal and pH requirements for activity, and in response to inhibitors. In the sporulating cell, an additional electrophoretic species of the enzyme was observed which could be partially converted to the principal form by treatment witlh Mn++; both forms of the enzyme required Mn++ for stabilization in solution as well as for activity. Inorganic pyrophosphatases obtained in a near homogeneous state from vegetative cells and spores of Bacillus subtilis proved to be indistinguishable when judged by a number of physical and functional criteria (H. Tono and A. Kornberg, J. Biol. Chem., in press). Similar results were obtained in recent studies of deoxyribonucleic acid (DNA) polymerase (1) and adenylate kinase in B. subtilis (J. A. Spudich and A. Kornberg, unpublished data), or nucleoside phosphorylase in B. cereius (R. Gardner and A. Kornberg, J. Biol. Chem., in press), and in studies of other enzymes as reviewed by Halvorson (2). From these results we derive the impression that a given spore enzyme and its vegetative-cell counterpart are the same despite their markedly different resistance to heat when tested in situ. For the long-range objective of defining the components and organization of a spore and the mechanisms

Properties and Developmental Roles of the Lysyl- and Tryptophanyl-Transfer Ribonucleic Acid Synthetases of Bacillus subtilis : Common Genetic Origin of the Corresponding Spore and Vegetative Enzymes

“…The two forms are genetically and structurally related, but depending on ionic conditions they exist in different aggregational states (5). Similar physical properties have been found for the glucose dehydrogenase of B. cereus (30) and the inorganic pyrophosphatase of' B. megaterium (38). In these cases, the evidence is consistent with the idea that the spore enzyme is a modified form of the vegetative enzyme.…”

Microbial inorganic pyrophosphatases.

“…Addition of Co2+ to the culture of Streptococcus faecalis increased the specific activity of PPase sevenfold (88). PPase activity was sinificantly increased also in spores of Bacillus subtilis grown in excess Mn2+ (112). The mechanisms by which the metals exert their effects are not known.…”

Reactivative Action of Ethylenediaminetetraacetic Acid or Dipicolinic Acid on Inactive Glucose Dehydrogenase Obtained from Heated Spores of Bacillus subtilis

“…Many workers (5,6,14,15) have reported that several enzymes in spores cannot be distinguished from those in vegetative cells. Therefore, the latter opinion has gained acceptance.…”