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. 1977 Aug;34(2):170–174. doi: 10.1128/aem.34.2.170-174.1977

Rapid spectrophotometric differentiation between glutathione-dependent and glutathione-independent gentisate and homogentisate pathways.

R L Crawford, T D Frick
PMCID: PMC242617  PMID: 907340

Abstract

A total of four pathways are known for the catabolism by microorganisms of gentisate (2,5-dihydroxybenzoate) and homogentisate (2,5-dihydroxyphenylacetate). Both of these dihydric phenols can be degraded by either a glutathione-dependent or a glutathione-independent reaction sequence. We found that it is not always possible to unequivocally assign glutathione dependence or independence to a particular catabolic sequence by using the well-established spectrophotometric assays at 330 nm (gentisate pathway) or 320 nm (homogentisate pathway). This paper reports a modification of the classical spectrophotometric assays that allowed an unequivocal differentiation between glutathion-dependent and glutathione-independent pathways, even when crude cell extracts contained significant quantities of cell-derived, reduced glutathione. This was accomplished by performing assays in the presence of an approximately 10(-3) M solution of the sulfhydryl-binding agent N-ethylmaleimide.

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Selected References

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

  1. CHAPMAN P. J., DAGLEY S. Oxidation of homogentistic acid by cell-free extracts of a vibrio. J Gen Microbiol. 1962 Jun;28:251–256. doi: 10.1099/00221287-28-2-251. [DOI] [PubMed] [Google Scholar]
  2. Crawford R. L. Degradation of 3-hydroxybenzoate by bacteria of the genus Bacillus. Appl Microbiol. 1975 Sep;30(3):439–444. doi: 10.1128/am.30.3.439-444.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Crawford R. L. Degradation of homogentisate by strains of Bacillus and Moraxella. Can J Microbiol. 1976 Feb;22(2):276–280. doi: 10.1139/m76-037. [DOI] [PubMed] [Google Scholar]
  4. Crawford R. L., Hutton S. W., Chapman P. J. Purification and properties of gentisate 1,2-dioxygenase from Moraxella osloensis. J Bacteriol. 1975 Mar;121(3):794–799. doi: 10.1128/jb.121.3.794-799.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Crawford R. L. Novel pathway for degradation of protocatechuic acid in Bacillus species. J Bacteriol. 1975 Feb;121(2):531–536. doi: 10.1128/jb.121.2.531-536.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Crawford R. L. Pathways of 4-hydroxybenzoate degradation among species of Bacillus. J Bacteriol. 1976 Jul;127(1):204–210. doi: 10.1128/jb.127.1.204-210.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dagley S. Catabolism of aromatic compounds by micro-organisms. Adv Microb Physiol. 1971;6(0):1–46. doi: 10.1016/s0065-2911(08)60066-1. [DOI] [PubMed] [Google Scholar]
  8. Hareland W. A., Crawford R. L., Chapman P. J., Dagley S. Metabolic function and properties of 4-hydroxyphenylacetic acid 1-hydroxylase from Pseudomonas acidovorans. J Bacteriol. 1975 Jan;121(1):272–285. doi: 10.1128/jb.121.1.272-285.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hopper D. J., Chapman P. J., Dagley S. Enzymic formation of D-malate. Biochem J. 1968 Dec;110(4):798–800. doi: 10.1042/bj1100798. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hopper D. J., Chapman P. J. Gentisic acid and its 3- and 4-methyl-substituted homologoues as intermediates in the bacterial degradation of m-cresol, 3,5-xylenol and 2,5-xylenol. Biochem J. 1971 Mar;122(1):19–28. doi: 10.1042/bj1220019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. KNOX W. E., EDWARDS S. W. The properties of maleylacetoacetate, the initial product of homogentisate oxidation in liver. J Biol Chem. 1955 Oct;216(2):489–498. [PubMed] [Google Scholar]
  12. LACK L. The enzymic oxidation of gentisic acid. Biochim Biophys Acta. 1959 Jul;34:117–123. doi: 10.1016/0006-3002(59)90239-2. [DOI] [PubMed] [Google Scholar]

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