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. 1970 Dec;104(3):1045–1051. doi: 10.1128/jb.104.3.1045-1051.1970

Localization in the Cell and Extraction of Alkaline Phosphatase from Bacillus subtilis

David A W Wood a,1, H Tristram a
PMCID: PMC248260  PMID: 16559076

Abstract

Study of protoplasts, lysed protoplasts, and cells treated with lysozyme in the absence of osmotic stabilizer suggested that the alkaline phosphatase (EC 3.1.3.1.) of Bacillus subtilis is located in the protoplasmic membrane. Cytochemical evidence in support of this view is presented. The enzyme protein was strongly bound to the membrane structure and could not be solubilized by a number of treatments known to release enzymes from membranes and other lipoprotein structures. Alkaline phosphatase was, however, solubilized by treatment of intact B. subtilis cells or isolated protoplasmic membranes with strong salt solutions at pH 7.2, suggesting that electrostatic forces are responsible for the association between membrane and enzyme protein. Dialysis of alkaline phosphatase solutions against buffer of low ionic strength resulted in precipitation of the enzyme.

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Selected References

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  1. BRENNER S., HORNE R. W. A negative staining method for high resolution electron microscopy of viruses. Biochim Biophys Acta. 1959 Jul;34:103–110. doi: 10.1016/0006-3002(59)90237-9. [DOI] [PubMed] [Google Scholar]
  2. Cashel M., Freese E. Excretion of alkaline phosphatase of Bacillus subtilis. Biochem Biophys Res Commun. 1964 Aug 11;16(6):541–544. doi: 10.1016/0006-291x(64)90189-5. [DOI] [PubMed] [Google Scholar]
  3. Coles N. W., Gross R. Liberation of surface-located penicillinase from Staphylococcus aureus. Biochem J. 1967 Mar;102(3):742–747. doi: 10.1042/bj1020742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Heppel L. A. Selective release of enzymes from bacteria. Science. 1967 Jun 16;156(3781):1451–1455. doi: 10.1126/science.156.3781.1451. [DOI] [PubMed] [Google Scholar]
  5. LUFT J. H. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. doi: 10.1083/jcb.9.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. MALAMY M., HORECKER B. L. The localization of alkaline phosphatase in E. coli K12. Biochem Biophys Res Commun. 1961 Jun 2;5:104–108. doi: 10.1016/0006-291x(61)90020-1. [DOI] [PubMed] [Google Scholar]
  7. Malveaux F. J., San Clemente C. L. Elution of loosely bound acid phosphatase from Staphylococcus aureus. Appl Microbiol. 1967 Jul;15(4):738–743. doi: 10.1128/am.15.4.738-743.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. NEALE S., TRISTRAM H. EFFECT OF O-METHYL-DL-THREONINE AND O-METHYL-DL-SERINE ON GROWTH AND PROTEIN SYNTHESIS IN ESCHERICHIA COLI. J Bacteriol. 1963 Dec;86:1241–1250. doi: 10.1128/jb.86.6.1241-1250.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Neu H. C., Heppel L. A. The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts. J Biol Chem. 1965 Sep;240(9):3685–3692. [PubMed] [Google Scholar]
  10. Neu H. C. The surface localization of penicillinases in Escherichia coli and Salmonella typhimurium. Biochem Biophys Res Commun. 1968 Jul 26;32(2):258–263. doi: 10.1016/0006-291x(68)90378-1. [DOI] [PubMed] [Google Scholar]
  11. SABATINI D. D., BENSCH K., BARRNETT R. J. Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J Cell Biol. 1963 Apr;17:19–58. doi: 10.1083/jcb.17.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Salton M. R. Structure and function of bacterial cell membranes. Annu Rev Microbiol. 1967;21:417–442. doi: 10.1146/annurev.mi.21.100167.002221. [DOI] [PubMed] [Google Scholar]
  13. TRISTRAM H. The adaptive degradation of L-histidine by Paracolobactrum aerogenoides. J Gen Microbiol. 1960 Dec;23:425–440. doi: 10.1099/00221287-23-3-425. [DOI] [PubMed] [Google Scholar]
  14. Takeda K., Tsugita A. Phosphoesterases of Bacillus subtilis. II. Crystallization and properties of alkaline phosphatase. J Biochem. 1967 Feb;61(2):231–241. doi: 10.1093/oxfordjournals.jbchem.a128535. [DOI] [PubMed] [Google Scholar]
  15. VERNON L. P., MANGUM J. H. Cytochromes of Bacillus megaterium and Bacillus subtilis. Arch Biochem Biophys. 1960 Sep;90:103–104. doi: 10.1016/0003-9861(60)90618-4. [DOI] [PubMed] [Google Scholar]
  16. VOELZ H. SITES OF ADENOSINE TRIPHOSPHATASE ACTIVITY IN BACTERIA. J Bacteriol. 1964 Oct;88:1196–1198. doi: 10.1128/jb.88.4.1196-1198.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Voelz H., Ortigoza R. O. Cytochemistry of phosphatases in Myxococcus xanthus. J Bacteriol. 1968 Oct;96(4):1357–1365. doi: 10.1128/jb.96.4.1357-1365.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. WEIBULL C. The isolation of protoplasts from Bacillus megaterium by controlled treatment with lysozyme. J Bacteriol. 1953 Dec;66(6):688–695. doi: 10.1128/jb.66.6.688-695.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Weimberg R., Orton W. L. Elution of Acid Phosphatase from the Cell Surface of Saccharomyces mellis by Potassium Chloride. J Bacteriol. 1965 Jul;90(1):82–94. doi: 10.1128/jb.90.1.82-94.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Yamaguchi T., Tamura G., Arima K. A method for obtaining cytochrome c from Bacillus megaterium and Bacillus subtilis. Biochim Biophys Acta. 1966 Aug 24;124(2):413–414. doi: 10.1016/0304-4165(66)90210-8. [DOI] [PubMed] [Google Scholar]

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