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. 1977 Mar 1;161(3):677–685. doi: 10.1042/bj1610677

Enzymic assays for isomers of 2,6-diaminopimelic acid in walls of Bacillus cereus and Bacillus megaterium.

A Day, P J White
PMCID: PMC1164555  PMID: 403912

Abstract

An enzymic assay for individual isomers (meso-, LL- and DD-) of 2,6-diaminopimelate was developed. The enzyme 2,6-diaminopimelate decarboxylase specifically attacked meso-diaminopimelate and was used to measure this isomer manometrically. The meso- and LL-isomers were measured together manometrically in a coupled assay with diaminopimelate decarboxylase and diaminopimelate epimerase (which converts LL-diaminopimelate into meso-diaminopimelate). The DD-isomer was not attacked by either enzyme and was measured, as residual diaminopimelate after the coupled assay, by a colorimetric method, which was also used to measure total diaminopimelate before enzymic treatments. The coupled enzymes were also used to prepare pure DD-isomer from chemically synthesized diaminopimelate. A mixture of diaminopimelate isomers was present in walls of four strains of Bacillus megaterium [in each about 75% (w/w) meso-, 18% LL- and 7% DD-] and in walls of two strains of Bacillus cereus (about 85% meso-, 8% LL- and 7% DD-). One strain of B. cereus contained at least 95% meso-diaminopimelate, with only traces of LL- and DD-isomers. Peptidoglycan from Escherichia coli was assayed as containing at least 95% meso-isomer. The proportion of isomers in the wall of a strain of B. megaterium remained constant after growth in a variety of different media.

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Selected References

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

  1. Bricas E., Ghuysen J. M., Dezélée P. The cell wall peptidoglycan of Bacillus megaterium KM. I. Studies on the stereochemistry of alpha, alpha'-diaminopimelic acid. Biochemistry. 1967 Aug;6(8):2598–2607. doi: 10.1021/bi00860a043. [DOI] [PubMed] [Google Scholar]
  2. DAVIS B. D. Biosynthetic interrelations of lysine, diaminopimelic acid, and threonine in mutants of Escherichia coli. Nature. 1952 Mar 29;169(4300):534–536. doi: 10.1038/169534a0. [DOI] [PubMed] [Google Scholar]
  3. DAVIS B. D., MINGIOLI E. S. Mutants of Escherichia coli requiring methionine or vitamin B12. J Bacteriol. 1950 Jul;60(1):17–28. doi: 10.1128/jb.60.1.17-28.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fiedler F., Kandler O. Die Aminosäuresequenz von 2,4-diaminobuttersäure enthaltenden Mureinen bei verschiedenen coryneformen Bakterien und Agromyces ramosus. Arch Mikrobiol. 1973;89(1):51–66. [PubMed] [Google Scholar]
  5. Grandgenett D. P., Stahly D. P. Repression of diaminopimelate decarboxylase by L-lysine in different Bacillus species. J Bacteriol. 1971 Mar;105(3):1211–1212. doi: 10.1128/jb.105.3.1211-1212.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Guinand M., Ghuysen J. M., Schleifer K. H., Kandler O. The peptidoglycan in walls of Butyribacterium rettgeri. Biochemistry. 1969 Jan;8(1):200–207. doi: 10.1021/bi00829a029. [DOI] [PubMed] [Google Scholar]
  7. HOARE D. S., WORK E. The stereoisomers of alpha epsilon-diaminopimelic acid. II. Their distribution in the bacterial order Actinomycetales and in certain Eubacteriales. Biochem J. 1957 Mar;65(3):441–447. doi: 10.1042/bj0650441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. LEVVY G. A., MCALLAN A. The N-acetylation and estimation of hexosamines. Biochem J. 1959 Sep;73:127–132. doi: 10.1042/bj0730127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Leive L., Davis B. D. The transport of diaminopimelate and cystine in Escherichia coli. J Biol Chem. 1965 Nov;240(11):4362–4369. [PubMed] [Google Scholar]
  11. MILNER H. W., LAWRENCE N. S., FRENCH C. S. Colloidal dispersion of chloroplast material. Science. 1950 Jun 9;111(2893):633–634. doi: 10.1126/science.111.2893.633. [DOI] [PubMed] [Google Scholar]
  12. Perkins H. R. 2,6-Diamino-3-hydroxypimelic acid in microbial cell wall mucopeptide. Nature. 1965 Nov 27;208(5013):872–873. doi: 10.1038/208872a0. [DOI] [PubMed] [Google Scholar]
  13. Perkins H. R. The configuration of 2,6-diamino-3-hydroxypimelic acid in microbial cell walls. Biochem J. 1969 Dec;115(4):797–805. doi: 10.1042/bj1150797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pitel D. W., Gilvarg C. Mucopeptide metabolism during growth and sporulation in Bacillus megaterium. J Biol Chem. 1970 Dec 25;245(24):6711–6717. [PubMed] [Google Scholar]
  15. SALTON M. R. Cell-wall amino-acids and amino-sugars. Nature. 1957 Aug 17;180(4581):338–339. doi: 10.1038/180338a0. [DOI] [PubMed] [Google Scholar]
  16. SCHOCHER A. J., BAYLEY S. T., WATSON R. W. Composition of purified mucopeptide from the wall of Aerobacter cloacae. Can J Microbiol. 1962 Feb;8:89–98. doi: 10.1139/m62-012. [DOI] [PubMed] [Google Scholar]
  17. Saleh F., White P. J. Use of auxotrophic mutants to isolate LL- or DD-isomers of 2,6-diaminopimelic acid. J Gen Microbiol. 1976 Oct;96(2):253–261. doi: 10.1099/00221287-96-2-253. [DOI] [PubMed] [Google Scholar]
  18. Schleifer K. H., Kandler O. Peptidoglycan types of bacterial cell walls and their taxonomic implications. Bacteriol Rev. 1972 Dec;36(4):407–477. doi: 10.1128/br.36.4.407-477.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Van Heijenoort J., Elbaz L., Dezélée P., Petit J. F., Bricas E., Ghuysen J. M. Structure of the meso-diaminopimelic acid containing peptidoglycans in Escherichia coli B and Bacillus megaterium KM. Biochemistry. 1969 Jan;8(1):207–213. doi: 10.1021/bi00829a030. [DOI] [PubMed] [Google Scholar]
  20. WHITE P. J., KELLY B. PURIFICATION AND PROPERTIES OF DIAMINOPIMELATE DECARBOXYLASE FROM ESCHERICHIA COLI. Biochem J. 1965 Jul;96:75–84. doi: 10.1042/bj0960075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. WORK E. Reaction of ninhydrin in acid solution with straight-chain amino acids containing two amino groups and its application to the estimation of alpha epsilon-diaminopimelic acid. Biochem J. 1957 Nov;67(3):416–423. doi: 10.1042/bj0670416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. White P. J., Lejeune B., Work E. Assay and properties of diaminopimelate epimerase from Bacillus megaterium. Biochem J. 1969 Jul;113(4):589–601. doi: 10.1042/bj1130589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. White P. J. The nutrition of Bacillus megaterium and Bacillus cereus. J Gen Microbiol. 1972 Aug;71(3):505–514. doi: 10.1099/00221287-71-3-505. [DOI] [PubMed] [Google Scholar]
  24. Wickus G. G., Strominger J. L. Penicillin-sensitive transpeptidation during peptidoglycan biosynthesis in cell-free preparations from Bacillus megaterium. I. Incorporation of free diaminopimelic acid into peptidoglycan. J Biol Chem. 1972 Sep 10;247(17):5297–5306. [PubMed] [Google Scholar]
  25. YAMAGUCHI T. COMPARISON OF THE CELL-WALL COMPOSITION OF MORPHOLOGICALLY DISTINCT ACTINOMYCETES. J Bacteriol. 1965 Feb;89:444–453. doi: 10.1128/jb.89.2.444-453.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]

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