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. 1971 Sep;107(3):704–711. doi: 10.1128/jb.107.3.704-711.1971

Isolation and Characterization of a Phosphatidylethanolamine-Deficient Mutant of Bacillus subtilis

James L Beebe a,1
PMCID: PMC246991  PMID: 4999411

Abstract

A mutant of Bacillus subtilis ATCC 6051 deficient in phosphatidylethanolamine, an important membrane lipid, was isolated by a combination of nitrosoguanidine mutagenesis and penicillin concentration of auxotrophs employing phosphatidylethanolamine as a supplement. The mutant was compared to the parent strain with regard to lipid composition, growth, osmotic fragility, and staining character and differed substantially in each category. In addition to scant amounts of phosphatidylethanolamine, the mutant contained phosphatidylglycerol, cardiolipin, lysyl phosphatidylglycerol, and diglucosyldiglyceride, though in amounts differing from those found in the parent strain. The mutant was unable to grow appreciably on synthetic media, had enhanced osmotic fragility of protoplasts, and resisted decolorization in staining.

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Selected References

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  1. AMES B. N., DUBIN D. T. The role of polyamines in the neutralization of bacteriophage deoxyribonucleic acid. J Biol Chem. 1960 Mar;235:769–775. [PubMed] [Google Scholar]
  2. BECKER B., LECHEVALIER M. P., LECHEVALIER H. A. CHEMICAL COMPOSITION OF CELL-WALL PREPARATIONS FROM STRAINS OF VARIOUS FORM-GENERA OF AEROBIC ACTINOMYCETES. Appl Microbiol. 1965 Mar;13:236–243. doi: 10.1128/am.13.2.236-243.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bishop D. G., Rutberg L., Samuelsson B. The chemical composition of the cytoplasmic membrane of Bacillus subtilis. Eur J Biochem. 1967 Nov;2(4):448–453. doi: 10.1111/j.1432-1033.1967.tb00158.x. [DOI] [PubMed] [Google Scholar]
  4. CROCKEN B. J., NYC J. F. PHOSPHOLIPID VARIATIONS IN MUTANT STRAINS OF NEUROSPORA CRASSA. J Biol Chem. 1964 Jun;239:1727–1730. [PubMed] [Google Scholar]
  5. Card G. L., Georgi C. E., Militzer W. E. Phospholipids from Bacillus stearothermophilus. J Bacteriol. 1969 Jan;97(1):186–192. doi: 10.1128/jb.97.1.186-192.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cronan J. E., Jr, Ray T. K., Vagelos P. R. Selection and characterization of an E. coli mutant defective in membrane lipid biosynthesis. Proc Natl Acad Sci U S A. 1970 Mar;65(3):737–744. doi: 10.1073/pnas.65.3.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Deierkauf F. A., Booij H. L. Changes in the phosphatide pattern of yeast cells in relation to active carbohydrate transport. Biochim Biophys Acta. 1968 Mar 1;150(2):214–225. doi: 10.1016/0005-2736(68)90165-x. [DOI] [PubMed] [Google Scholar]
  8. FOLCH J., ASCOLI I., LEES M., MEATH J. A., LeBARON N. Preparation of lipide extracts from brain tissue. J Biol Chem. 1951 Aug;191(2):833–841. [PubMed] [Google Scholar]
  9. GARY N. D., BARD R. C. Effect of nutrition on the growth and metabolism of Bacillus subtilis. J Bacteriol. 1952 Oct;64(4):501–512. doi: 10.1128/jb.64.4.501-512.1952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Henning U., Dennert G., Rehn K., Deppe G. Effects of oleate starvation in a fatty acid auxotroph of Escherichia coli K-12. J Bacteriol. 1969 May;98(2):784–796. doi: 10.1128/jb.98.2.784-796.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ikawa M. Bacterial phosphatides and natural relationships. Bacteriol Rev. 1967 Mar;31(1):54–64. doi: 10.1128/br.31.1.54-64.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kappy M. S., Metzenberg R. L. Multiple alterations in metabolite uptake in a mutant of Neurospora crassa. J Bacteriol. 1967 Nov;94(5):1629–1637. doi: 10.1128/jb.94.5.1629-1637.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Keith A. D., Resnick M. R., Haley A. B. Fatty acid desaturase mutants of Saccharomyces cerevisiae. J Bacteriol. 1969 May;98(2):415–420. doi: 10.1128/jb.98.2.415-420.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kito M., Lubin M., Pizer L. I. A mutant of Escherichia coli defective in phosphatidic acid synthesis. Biochem Biophys Res Commun. 1969 Feb 21;34(4):454–458. doi: 10.1016/0006-291x(69)90403-3. [DOI] [PubMed] [Google Scholar]
  15. LEDERBERG J., LEDERBERG E. M. Replica plating and indirect selection of bacterial mutants. J Bacteriol. 1952 Mar;63(3):399–406. doi: 10.1128/jb.63.3.399-406.1952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Milner L. S., Kaback H. R. The role of phosphatidylglycerol in the vectorial phosphorylation of sugar by isolated bacterial membrane preparations. Proc Natl Acad Sci U S A. 1970 Mar;65(3):683–690. doi: 10.1073/pnas.65.3.683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mindich L. Membrane synthesis in Bacillus subtilis. I. Isolation and properties of strains bearing mutations in glycerol metabolism. J Mol Biol. 1970 Apr 28;49(2):415–432. doi: 10.1016/0022-2836(70)90254-8. [DOI] [PubMed] [Google Scholar]
  18. Mindich L. Membrane synthesis in Bacillus subtilis. II. Integration of membrane proteins in the absence of lipid synthesis. J Mol Biol. 1970 Apr 28;49(2):433–439. doi: 10.1016/0022-2836(70)90255-x. [DOI] [PubMed] [Google Scholar]
  19. RADIN N. S., LAVIN F. B., BROWN J. R. Determination of cerebrosides. J Biol Chem. 1955 Dec;217(2):789–796. [PubMed] [Google Scholar]
  20. RENKONEN O. Determination of glycerol in phosphatides. Biochim Biophys Acta. 1962 Jan 29;56:367–369. doi: 10.1016/0006-3002(62)90580-2. [DOI] [PubMed] [Google Scholar]
  21. Resnick M. A., Mortimer R. K. Unsaturated fatty acid mutants of Saccharomyces cerevisiae. J Bacteriol. 1966 Sep;92(3):597–600. doi: 10.1128/jb.92.3.597-600.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. SNYDER F., STEPHENS N. A simplified spectrophotometric determination of ester groups in lipids. Biochim Biophys Acta. 1959 Jul;34:244–245. doi: 10.1016/0006-3002(59)90255-0. [DOI] [PubMed] [Google Scholar]
  23. Sanger F. The free amino groups of insulin. Biochem J. 1945;39(5):507–515. doi: 10.1042/bj0390507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Silbert D. F. Arrangement of fatty acyl groups in phosphatidylethanolamine from a fatty acid auxotroph of Escherichia coli. Biochemistry. 1970 Sep 1;9(18):3631–3640. doi: 10.1021/bi00820a021. [DOI] [PubMed] [Google Scholar]
  25. Silbert D. F., Ruch F., Vagelos P. R. Fatty acid replacements in a fatty acid auxotroph of Escherichia coli. J Bacteriol. 1968 May;95(5):1658–1665. doi: 10.1128/jb.95.5.1658-1665.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Silbert D. F., Vagelos P. R. Fatty acid mutant of E. coli lacking a beta-hydroxydecanoyl thioester dehydrase. Proc Natl Acad Sci U S A. 1967 Oct;58(4):1579–1586. doi: 10.1073/pnas.58.4.1579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Urakami C., Umetani K. Compositions of phosphatides from Bacillus natto at various growth phases. Biochim Biophys Acta. 1968 Sep 2;164(1):64–71. [PubMed] [Google Scholar]
  28. VORBECK M. L., MARINETTI G. V. SEPARATION OF GLYCOSYL DIGLYCERIDES FROM PHOSPHATIDES USING SILICIC ACID COLUMN CHROMATOGRAPHY. J Lipid Res. 1965 Jan;6:3–6. [PubMed] [Google Scholar]
  29. 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]
  30. WELLS M. A., DITTMER J. C. THE USE OF SEPHADEX FOR THE REMOVAL OF NONLIPID CONTAMINANTS FROM LIPID EXTRACTS. Biochemistry. 1963 Nov-Dec;2:1259–1263. doi: 10.1021/bi00906a015. [DOI] [PubMed] [Google Scholar]
  31. White D. C., Frerman F. E. Extraction, characterization, and cellular localization of the lipids of Staphylococcus aureus. J Bacteriol. 1967 Dec;94(6):1854–1867. doi: 10.1128/jb.94.6.1854-1867.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wisnieski B. J., Keith A. D., Resnick M. R. Double-bond requirement in a fatty acid desaturase mutant of Saccharomyces cerevisiae. J Bacteriol. 1970 Jan;101(1):160–165. doi: 10.1128/jb.101.1.160-165.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. den Kamp J. A., Redai I., van Deenen L. L. Phospholipid composition of Bacillus subtilis. J Bacteriol. 1969 Jul;99(1):298–303. doi: 10.1128/jb.99.1.298-303.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]

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