Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1968 Apr;95(4):1327–1334. doi: 10.1128/jb.95.4.1327-1334.1968

Initiation of Bacterial Spore Germination

J C Vary a,1, H O Halvorson a
PMCID: PMC315090  PMID: 4296513

Abstract

To investigate the problem of initiation in bacterial spore germination, we isolated, from extracts of dormant spores of Bacillus cereus strain T and B. licheniformis, a protein that initiated spore germination when added to a suspension of heat-activated spores. The optimal conditions for initiatory activity of this protein (the initiator) were 30 C in 0.01 to 0.04 m NaCl and 0.01 m tris(hydroxymethyl)aminomethane (pH 8.5). The initiator was inhibited by phosphate but required two co-factors, l-alanine (1/7 of Km for l-alanine-inhibited germination) and nicotinamide adenine dinucleotide (1.25 × 10−4m). In the crude extract, the initiator activity was increased 3.5-fold by heating the extract at 65 C for 10 min, but the partially purified initiator preparation was completely heat-sensitive (65 C for 5 min). Heat stability could be conferred on the purified initiator by adding 10−3m dipicolinic acid. A fractionation of this protein that excluded l-alanine dehydrogenase and adenosine deaminase from the initiator activity was developed. The molecular weight of the initiator was estimated as 7 × 104. The kinetics of germination in the presence of initiator were examined at various concentrations of l-alanine and nicotinamide adenine dinucleotide.

Full text

PDF
1334

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andrews P. The gel-filtration behaviour of proteins related to their molecular weights over a wide range. Biochem J. 1965 Sep;96(3):595–606. doi: 10.1042/bj0960595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. CHURCH B. D., HALVORSON H., HALVORSON H. O. Studies on spore germination: its independence from alanine racemase activity. J Bacteriol. 1954 Oct;68(4):393–399. doi: 10.1128/jb.68.4.393-399.1954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. CLELAND W. W. The kinetics of enzyme-catalyzed reactions with two or more substrates or products. I. Nomenclature and rate equations. Biochim Biophys Acta. 1963 Jan 8;67:104–137. doi: 10.1016/0006-3002(63)91800-6. [DOI] [PubMed] [Google Scholar]
  4. Gould G. W., King W. L. Role of amino-acids in the germination of bacterial spores by subtilisn. Nature. 1966 Sep 24;211(5056):1431–1432. doi: 10.1038/2111431a0. [DOI] [PubMed] [Google Scholar]
  5. Hachisuka Y., Tochikubo K., Yokoi Y., Murachi T. The action of dipicolinic acid and its chemical analogues on the heat stability of glucose dehydrogenase of Bacillus subtilis spores. J Biochem. 1967 May;61(5):659–661. doi: 10.1093/oxfordjournals.jbchem.a128598. [DOI] [PubMed] [Google Scholar]
  6. Halvorson H. O., Vary J. C., Steinberg W. Developmental changes during the formation and breaking of the dormant state in bacteria. Annu Rev Microbiol. 1966;20:169–188. doi: 10.1146/annurev.mi.20.100166.001125. [DOI] [PubMed] [Google Scholar]
  7. JANSSEN F. W., LUND A. J., ANDERSON L. E. Colorimetric assay for dipicolinic acid in bacterial spores. Science. 1958 Jan 3;127(3288):26–27. doi: 10.1126/science.127.3288.26. [DOI] [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. MARTIN R. G., AMES B. N. A method for determining the sedimentation behavior of enzymes: application to protein mixtures. J Biol Chem. 1961 May;236:1372–1379. [PubMed] [Google Scholar]
  10. MCCORMICK N. G. KINETICS OF SPORE GERMINATION. J Bacteriol. 1965 May;89:1180–1185. doi: 10.1128/jb.89.5.1180-1185.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mishiro Y., Ochi M. Effect of dipicolinate on the heat denaturation of proteins. Nature. 1966 Sep 10;211(5054):1190–1190. doi: 10.1038/2111190a0. [DOI] [PubMed] [Google Scholar]
  12. Moberly B. J., Shafa F., Gerhardt P. Structural details of anthrax spores during stages of transformation into vegetative cells. J Bacteriol. 1966 Jul;92(1):220–228. doi: 10.1128/jb.92.1.220-228.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. STRANGE R. E., DARK F. A. A cell-wall lytic enzyme associated with spores of Bacillus species. J Gen Microbiol. 1957 Feb;16(1):236–249. doi: 10.1099/00221287-16-1-236. [DOI] [PubMed] [Google Scholar]
  14. Sierra G. Germination of bacterial endospores with subtilopeptidases. Can J Microbiol. 1967 May;13(5):489–501. doi: 10.1139/m67-064. [DOI] [PubMed] [Google Scholar]
  15. VARY J. C., HALVORSON H. O. KINETICS OF GERMINATION OF BACILLUS SPORES. J Bacteriol. 1965 May;89:1340–1347. doi: 10.1128/jb.89.5.1340-1347.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Vallee B. L., Hoch F. L. ZINC, A COMPONENT OF YEAST ALCOHOL DEHYDROGENASE. Proc Natl Acad Sci U S A. 1955 Jun 15;41(6):327–338. doi: 10.1073/pnas.41.6.327. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES