Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1982 Mar;149(3):916–922. doi: 10.1128/jb.149.3.916-922.1982

Genetic and biochemical analyses of pantothenate biosynthesis in Escherichia coli and Salmonella typhimurium.

J E Cronan Jr, K J Littel, S Jackowski
PMCID: PMC216478  PMID: 7037743

Abstract

Pantothenate (pan) auxotrophs of Escherichia coli K-12 and Salmonella typhimurium LT2 were characterized by enzymatic and genetic analyses. The panB mutants of both organisms and the pan-6 ("panA") mutant of S. typhimurium are deficient in ketopantoate hydroxymethyltransferase, whereas the panC mutants lack pantothenate synthetase. panD mutants of E. coli K-12 were previously shown to be deficient in aspartate 1-decarboxylase. All mutants showed only a single enzyme defect. The finding that the pan-6 mutant was deficient in ketopantoate hydroxymethyltransferase indicates that the genetic lesion is a panB allele. The pan-6 mutant therefore is deficient in the utilization of alpha-ketoisovalerate rather than the synthesis of alpha-ketoisovalerate, as originally proposed. The order of the pan genes of E. coli K-12 was determined by phage P1-mediated three-factor crosses. The clockwise order was found to be aceF panB panD panC tonA on the genetic map of E. coli K-12. The three-factor crosses were greatly facilitated by use of a closely linked Tn10 transposon as the outside marker. We also found that supplementation of E. coli K-12 auxotrophs with a high concentration of pantothenate or beta-alanine increased the intracellular coenzyme A level two- to threefold above the normal level. Supplementation with pantoate or ketopantoate resulted in smaller increases.

Full text

PDF
916

Selected References

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

  1. Alberts A. W., Vagelos P. R. Acyl carrier protein. 8. Studies of acyl carrier protein and coenzyme A in Escherichia coli pantothenate or betaalanine auxotrophs. J Biol Chem. 1966 Nov 25;241(22):5201–5204. [PubMed] [Google Scholar]
  2. Allred J. B., Guy D. G. Determination of coenzyme A and acetyl CoA in tissue extracts. Anal Biochem. 1969 May;29(2):293–299. doi: 10.1016/0003-2697(69)90312-1. [DOI] [PubMed] [Google Scholar]
  3. BRITTEN R. J., McCLURE F. T. The amino acid pool in Escherichia coli. Bacteriol Rev. 1962 Sep;26:292–335. doi: 10.1128/br.26.3.292-335.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. BROWN G. M. The metabolism of pantothenic acid. J Biol Chem. 1959 Feb;234(2):370–378. [PubMed] [Google Scholar]
  5. Bachmann B. J., Low K. B. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. doi: 10.1128/mr.44.1.1-56.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clark D. P., Cronan J. E., Jr Acetaldehyde coenzyme A dehydrogenase of Escherichia coli. J Bacteriol. 1980 Oct;144(1):179–184. doi: 10.1128/jb.144.1.179-184.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cronan J. E., Jr Beta-alanine synthesis in Escherichia coli. J Bacteriol. 1980 Mar;141(3):1291–1297. doi: 10.1128/jb.141.3.1291-1297.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cronan J. E., Jr, Silbert D. F., Wulff D. L. Mapping of the fabA locus for unsaturated fatty acid biosynthesis in Escherichia coli. J Bacteriol. 1972 Oct;112(1):206–211. doi: 10.1128/jb.112.1.206-211.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jackowski S., Rock C. O. Regulation of coenzyme A biosynthesis. J Bacteriol. 1981 Dec;148(3):926–932. doi: 10.1128/jb.148.3.926-932.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Loewen P. C. Effect of glutathione deficiency on the pool of CoA-glutathione mixed disulfide in Escherichia coli. Can J Biochem. 1981 May;59(5):379–382. doi: 10.1139/o81-052. [DOI] [PubMed] [Google Scholar]
  11. Loewen P. C. Levels of coenzyme A--glutathione mixed disulfide in Escherichia coli. Can J Biochem. 1978 Jul;56(7):753–759. doi: 10.1139/o78-113. [DOI] [PubMed] [Google Scholar]
  12. MAAS W. K., DAVIS B. D. Pantothenate studies. I. Interference by D-serine and L-aspartic acid with pantothenate synthesis in Escherichia coli. J Bacteriol. 1950 Dec;60(6):733–745. doi: 10.1128/jb.60.6.733-745.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. MAAS W. K. Pantothenate studies. III. Description of the extracted pantothenate-synthesizing enzyme of Escherichia coli. J Biol Chem. 1952 Sep;198(1):23–32. [PubMed] [Google Scholar]
  14. Miyatake K., Nakano Y., Kitaoka S. Pantothenate synthetase from Escherichia coli [D-pantoate: beta-alanine ligase (AMP-forming), EC 6.3.2.1]. Methods Enzymol. 1979;62:215–219. doi: 10.1016/0076-6879(79)62221-8. [DOI] [PubMed] [Google Scholar]
  15. Mäntsälä P. Some characteristics and control of pantothenate transport in Escherichia coli U-5-41. Acta Chem Scand. 1973;27(2):445–452. doi: 10.3891/acta.chem.scand.27-0445. [DOI] [PubMed] [Google Scholar]
  16. Ortega M. V., Cárdenas A., Ubiera D. panD, a new chromosomal locus of Salmonella typhimurium for the biosynthesis of beta-alanine. Mol Gen Genet. 1975 Sep 29;140(2):159–164. doi: 10.1007/BF00329783. [DOI] [PubMed] [Google Scholar]
  17. Polacco M. L., Cronan J. E., Jr A mutant of Escherichia coli conditionally defective in the synthesis of holo-[acyl carrier protein]. J Biol Chem. 1981 Jun 10;256(11):5750–5754. [PubMed] [Google Scholar]
  18. Powers S. G., Snell E. E. Ketopantoate hydroxymethyltransferase. II. Physical, catalytic, and regulatory properties. J Biol Chem. 1976 Jun 25;251(12):3786–3793. [PubMed] [Google Scholar]
  19. Sanderson K. E., Hartman P. E. Linkage map of Salmonella typhimurium, edition V. Microbiol Rev. 1978 Jun;42(2):471–519. doi: 10.1128/mr.42.2.471-519.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Teller J. H., Powers S. G., Snell E. E. Ketopantoate hydroxymethyltransferase. I. Purification and role in pantothenate biosynthesis. J Biol Chem. 1976 Jun 25;251(12):3780–3785. [PubMed] [Google Scholar]
  21. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  22. Van Den Bosch H., Williamson J. R., Vagelos P. R. Localization of acyl carrier protein in Escherichia coli. Nature. 1970 Oct 24;228(5269):338–341. doi: 10.1038/228338a0. [DOI] [PubMed] [Google Scholar]
  23. Wilken D. R., King H. L., Jr, Dyar R. E. Ketopantoic acid and ketopantoyl lactone reductases. Stereospecificity of transfer of hydrogen from reduced nicotinamide adenine dinucleotide phosphate. J Biol Chem. 1975 Mar 25;250(6):2311–2314. [PubMed] [Google Scholar]
  24. Wilken D. R., King H. L., Jr, Dyar R. E. Ketopantoyl lactone reductases. Methods Enzymol. 1979;62:209–215. doi: 10.1016/0076-6879(79)62220-6. [DOI] [PubMed] [Google Scholar]
  25. Williamson J. M., Brown G. M. Purification and properties of L-Aspartate-alpha-decarboxylase, an enzyme that catalyzes the formation of beta-alanine in Escherichia coli. J Biol Chem. 1979 Aug 25;254(16):8074–8082. [PubMed] [Google Scholar]

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

RESOURCES