Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1990 Mar;56(3):724–729. doi: 10.1128/aem.56.3.724-729.1990

Assay for the enzymatic conversion of indoleacetic acid to 3-methylindole in a ruminal Lactobacillus species.

D C Honeyfield 1, J R Carlson 1
PMCID: PMC183413  PMID: 2317043

Abstract

An assay to measure the rate of enzymatic formation of 3-methylindole (3MI) from indoleacetic acid (IAA) in Lactobacillus sp. strain 11201 was developed. The reaction mixture contained 50 micrograms of microbial protein per ml (range, 25 to 100 mg/ml), essential low-molecular-weight reaction ingredients, and radiolabeled IAA as substrate (range, 0 to 2 mM IAA). The reaction was anaerobic for 25 min at 39 degrees C. The apparent Michaelis-Menten constants were: Km, 0.14 mM IAA; and Vmax, 64 nmol 3MI.mg-1.min-1. The inhibitors avidin, aminopterin, and EDTA had no effect on the 3MI-forming enzyme. Dithionite stimulated the 3MI-forming enzyme. The product of the reaction, 3MI, acted as a noncompetitive inhibitor of the enzyme. Enzyme activity was associated with the cell wall fraction after sonication; treatment with the French press; or treatment with detergents, proteolytic enzymes, and EDTA.

Full text

PDF
724

Images in this article

726Image
on p.726

Selected References

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

  1. BENTLEY R., THIESSEN C. P. Cisaconitic decarboxylase. Science. 1955 Aug 19;122(3164):330–330. doi: 10.1126/science.122.3164.330. [DOI] [PubMed] [Google Scholar]
  2. Bray T. M., Magnuson J. A., Carlson J. R. Nuclear magnetic resonance studies of lecithin-skatole interaction. J Biol Chem. 1974 Feb 10;249(3):914–918. [PubMed] [Google Scholar]
  3. Bray T. M., Sandberg H. E., Carlson J. R. An EPR study of structural perturbations induced by methylindole in the protein and lipid regions of erythrocyte membranes. Biochim Biophys Acta. 1975 Apr 8;382(4):534–541. doi: 10.1016/0005-2736(75)90220-5. [DOI] [PubMed] [Google Scholar]
  4. Breeze R. G., Carlson J. R. Chemical-induced lung injury in domestic animals. Adv Vet Sci Comp Med. 1982;26:201–231. [PubMed] [Google Scholar]
  5. Buswell J. A., Ander P., Pettersson B., Eriksson K. E. Oxidative decarboxylation of vanillic acid by Sporotrichum pulverulentum. FEBS Lett. 1979 Jul 1;103(1):98–101. doi: 10.1016/0014-5793(79)81258-2. [DOI] [PubMed] [Google Scholar]
  6. Coleman R. Membrane-bound enzymes and membrane ultrastructure. Biochim Biophys Acta. 1973 Apr 3;300(1):1–30. doi: 10.1016/0304-4157(73)90010-5. [DOI] [PubMed] [Google Scholar]
  7. D'Ari L., Barker H. A. p-Cresol formation by cell-free extracts of Clostridium difficile. Arch Microbiol. 1985 Dec;143(3):311–312. doi: 10.1007/BF00411256. [DOI] [PubMed] [Google Scholar]
  8. Dimroth P. Sodium ion transport decarboxylases and other aspects of sodium ion cycling in bacteria. Microbiol Rev. 1987 Sep;51(3):320–340. doi: 10.1128/mr.51.3.320-340.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Evans W. C. Biochemistry of the bacterial catabolism of aromatic compounds in anaerobic environments. Nature. 1977 Nov 3;270(5632):17–22. doi: 10.1038/270017a0. [DOI] [PubMed] [Google Scholar]
  10. FINKLE B. J., LEWIS J. C., CORSE J. W., LUNDIN R. E. Enzyme reactions with phenolic compounds: formation of hydroxystyrenes through the decarboxylation of 4-hydroxycinnamic acids by Aerobacter. J Biol Chem. 1962 Sep;237:2926–2931. [PubMed] [Google Scholar]
  11. Fellers C. R., Clough R. W. INDOL AND SKATOL DETERMINATION IN BACTERIAL CULTURES. J Bacteriol. 1925 Mar;10(2):105–133. doi: 10.1128/jb.10.2.105-133.1925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hickey M. W., Hillier A. J., Jago G. R. Metabolism of pyruvate and citrate in lactobacilli. Aust J Biol Sci. 1983;36(5-6):487–496. doi: 10.1071/bi9830487. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Lindsay R. F., Priest F. G. Decarboxylation of substituted cinnamic acids by enterobacteria: the influence on beer flavour. J Appl Bacteriol. 1975 Oct;39(2):181–187. doi: 10.1111/j.1365-2672.1975.tb00560.x. [DOI] [PubMed] [Google Scholar]
  15. NOLLER E. C., HARTSELL S. E. Bacteriolysis of Enterobacteriaceae. I. Lysis by four lytic systems utilizing lysozyme. J Bacteriol. 1961 Mar;81:482–491. doi: 10.1128/jb.81.3.482-491.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. ROSENBERGER R. F. Obligate anaerobes which form skatole. J Bacteriol. 1959 Apr;77(4):517–517. doi: 10.1128/jb.77.4.517-517.1959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Recsei P. A., Snell E. E. Pyruvoyl enzymes. Annu Rev Biochem. 1984;53:357–387. doi: 10.1146/annurev.bi.53.070184.002041. [DOI] [PubMed] [Google Scholar]
  18. Sleat R., Robinson J. P. The bacteriology of anaerobic degradation of aromatic compounds. J Appl Bacteriol. 1984 Dec;57(3):381–394. doi: 10.1111/j.1365-2672.1984.tb01404.x. [DOI] [PubMed] [Google Scholar]
  19. Spray R. S. Three New Species of the Genus Clostridium. J Bacteriol. 1948 Jun;55(6):839–842. doi: 10.1128/jb.55.6.839-842.1948. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Taylor B. F., Ribbons D. W. Bacterial Decarboxylation of o-Phthalic Acids. Appl Environ Microbiol. 1983 Dec;46(6):1276–1281. doi: 10.1128/aem.46.6.1276-1281.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Thomas P. S., Farquhar M. N. Specific measurement of DNA in nuclei and nucleic acids using diaminobenzoic acid. Anal Biochem. 1978 Aug 15;89(1):35–44. doi: 10.1016/0003-2697(78)90724-8. [DOI] [PubMed] [Google Scholar]
  22. WHITING G. C., CARR J. G. Metabolism of cinnamic acid and hydroxy-cinnamic acids by Lactobacillus pastorianus var. quinicus. Nature. 1959 Oct 31;184(Suppl 18):1427–1428. doi: 10.1038/1841427a0. [DOI] [PubMed] [Google Scholar]
  23. Ward L. A., Johnson K. A., Robinson I. M., Yokoyama M. T. Isolation from swine feces of a bacterium which decarboxylates p-hydroxyphenylacetic acid to 4-methylphenol (p-cresol). Appl Environ Microbiol. 1987 Jan;53(1):189–192. doi: 10.1128/aem.53.1.189-192.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Yokoyama M. T., Carlson J. R., Holdeman L. V. Isolation and characteristics of a skatole-producing Lactobacillus sp. from the bovine rumen. Appl Environ Microbiol. 1977 Dec;34(6):837–842. doi: 10.1128/aem.34.6.837-842.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Yokoyama M. T., Carlson J. R. Production of Skatole and para-Cresol by a Rumen Lactobacillus sp. Appl Environ Microbiol. 1981 Jan;41(1):71–76. doi: 10.1128/aem.41.1.71-76.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES