Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1989 Aug 15;262(1):303–312. doi: 10.1042/bj2620303

Purification and characterization of 4-methylmuconolactone methyl-isomerase, a novel enzyme of the modified 3-oxoadipate pathway in nocardioform actinomycetes.

N C Bruce 1, R B Cain 1, D H Pieper 1, K H Engesser 1
PMCID: PMC1133261  PMID: 2818569

Abstract

The novel enzyme 4-methyl-2-enelactone methyl-isomerase was detected in, and purified to electrophoretic homogeneity from, p-toluate-grown cells of Rhodococcus rhodocrous N75, a nocardioform actinomycete. The enzyme was very thermostable and had a native Mr of 75,500; as the monomer had an Mr of 17,000, the enzyme is probably tetrameric. The new isomerase is highly specific with respect to its lactone substrate, only accepting (+)-(4S)-4-methylmuconolactone (4-carboxymethyl-4-methylbut-2-en-1,4-olide), and the putative isomerization reaction intermediate 1-methylbislactone ((-)-1-methyl-3,7-dioxo-2,6-dioxabicyclo-[3.3.0]octane) as substrates, and yielding (-)-(4S)-3-methylmuconolactone (4-carboxymethyl-3-methylbut-2-en-1,4-olide) as product. Some other lactone analogues acted as competitive inhibitors. Our data suggest that the isomerization does not involve actual methyl migration, but proceeds via the 1-methybislactone.

Full text

PDF
309

Selected References

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

  1. Andrews P. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. doi: 10.1042/bj0910222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cain R. B., Bilton R. F., Darrah J. A. The metabolism of aromatic acids by micro-organisms. Metabolic pathways in the fungi. Biochem J. 1968 Aug;108(5):797–828. doi: 10.1042/bj1080797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cain R. B. The metabolism of protocatechuic acid by certain micro-organisms. A reassessment of the evidence for the participation of 2:6-dioxa-3:7-dioxobicyclo[3:3:0]octane as an intermediate. Biochem J. 1961 May;79(2):312–316. doi: 10.1042/bj0790312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Catelani D., Fiecchi A., Galli E. Dextro-gamma-carboxymethyl-gamma-methyl-delta-alpha-butenolide. A 1,2-ring-fission product of 4-methylcatechol by Pseudomonas desmolyticum. Biochem J. 1971 Jan;121(1):89–92. doi: 10.1042/bj1210089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. ELSDEN S. R., PEEL J. L. Metabolism of carbohydrates and related compounds. Annu Rev Microbiol. 1958;12:145–202. doi: 10.1146/annurev.mi.12.100158.001045. [DOI] [PubMed] [Google Scholar]
  6. GROSS S. R., GAFFORD R. D., TATUM E. L. The metabolism of protocatechuic acid by Neurospora. J Biol Chem. 1956 Apr;219(2):781–796. [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Murray K., Duggleby C. J., Sala-Trepat J. M., Williams P. A. The metabolism of benzoate and methylbenzoates via the meta-cleavage pathway by Pseudomonas arvilla mt-2. Eur J Biochem. 1972 Jul 24;28(3):301–310. doi: 10.1111/j.1432-1033.1972.tb01914.x. [DOI] [PubMed] [Google Scholar]
  9. Powlowski J. B., Dagley S. beta-Ketoadipate pathway in Trichosporon cutaneum modified for methyl-substituted metabolites. J Bacteriol. 1985 Sep;163(3):1126–1135. doi: 10.1128/jb.163.3.1126-1135.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Sala-Trepat J. M., Murray K., Williams P. A. The metabolic divergence in the meta cleavage of catechols by Pseudomonas putida NCIB 10015. Physiological significance and evolutionary implications. Eur J Biochem. 1972 Jul 24;28(3):347–356. doi: 10.1111/j.1432-1033.1972.tb01920.x. [DOI] [PubMed] [Google Scholar]
  11. Stanier R. Y., Ornston L. N. The beta-ketoadipate pathway. Adv Microb Physiol. 1973;9(0):89–151. [PubMed] [Google Scholar]
  12. Worsey M. J., Franklin F. C., Williams P. A. Regulation of the degradative pathway enzymes coded for by the TOL plasmid (pWWO) from Pseudomonas putida mt-2. J Bacteriol. 1978 Jun;134(3):757–764. doi: 10.1128/jb.134.3.757-764.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES