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. 1978 Nov 1;175(2):649–658. doi: 10.1042/bj1750649

P-cresol and 3,5-xylenol methylhydroxylases in Pseudomonas putida N.C.I.B. 9896.

M J Keat, D J Hopper
PMCID: PMC1186115  PMID: 743215

Abstract

Pseudomonas putida N.C.I.B. 9869, when grown on 3,5-xylenol, hydroxylates the methyl groups on 3,5-xylenol and on p-cresol by two different enzymes. 3,5-Xylenol methylhydroxylase, studied only in relatively crude extracts, requires NADH, is not active with p-cresol and is inhibited by cyanide, but not by CO. The p-cresol methylhydroxylase requires an electron acceptor and will act under anaerobic conditions. It was purified and is a flavocytochrome c of mol.wt. approx. 114,000 consisting of two subunits of equal size. The enzyme catalyses the hydroxylation of p-cresol (Km 16 micron) and the further oxidation of product, p-hydroxybenzyl alcohol (Km 27 micron) to p-hydroxybenzaldehyde. A different p-cresol methylhydroxylase of the flavocytochrome c type is induced by growth on p-cresol. It too was purified and has mol.wt. approx. 100,000, and again consisted of two equal-size subunits. The Km for p=cresol 3.6 micron and for p=hydroxybenzyl alcohol, 15 micron.

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Selected References

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  1. Andrews P. The gel-filtration behaviour of proteins related to their molecular weights over a wide range. Biochem J. 1965 Sep;96(3):595–606. doi: 10.1042/bj0960595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Byrne G. A. The separation of 2,4-dinitrophenylhydrazones by thin-layer chromatography. J Chromatogr. 1965 Dec;20(3):528–540. doi: 10.1016/s0021-9673(01)97455-2. [DOI] [PubMed] [Google Scholar]
  3. GHOLSON R. K., BAPTIST J. N., COON M. J. HYDROCARBON OXIDATION BY A BACTERIAL ENZYME SYSTEM. II. COFACTOR REQUIREMENTS FOR OCTANOL FORMATION FROM OCTANE. Biochemistry. 1963 Sep-Oct;2:1155–1159. doi: 10.1021/bi00905a043. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Hopper D. J., Taylor D. G. Pathways for the degradation of m-cresol and p-cresol by Pseudomonas putida. J Bacteriol. 1975 Apr;122(1):1–6. doi: 10.1128/jb.122.1.1-6.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hopper D. J., Taylor D. G. The purification and properties of p-cresol-(acceptor) oxidoreductase (hydroxylating), a flavocytochrome from Pseudomonas putida. Biochem J. 1977 Oct 1;167(1):155–162. doi: 10.1042/bj1670155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hopper D. J. The hydroxylation of P-cresol and its conversion to P-hydroxybenzaldehyde in Pseudomonas putida. Biochem Biophys Res Commun. 1976 Mar 22;69(2):462–468. doi: 10.1016/0006-291x(76)90544-1. [DOI] [PubMed] [Google Scholar]
  8. Keat M. J., Hopper D. J. Aromatic-alcohol dehydrogenases from Pseudomonas putida N.C.I.B. 9869. Biochem Soc Trans. 1976;4(3):494–495. doi: 10.1042/bst0040494. [DOI] [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. McKenna E. J., Coon M. J. Enzymatic omega-oxidation. IV. Purification and properties of the omega-hydroxylase of Pseudomonas oleovorans. J Biol Chem. 1970 Aug 10;245(15):3882–3889. [PubMed] [Google Scholar]
  11. ROSENBERGER R. F., ELSDEN S. R. The yields of Streptococcus faecalis grown in continuous culture. J Gen Microbiol. 1960 Jun;22:726–739. doi: 10.1099/00221287-22-3-726. [DOI] [PubMed] [Google Scholar]
  12. Ruettinger R. T., Griffith G. R., Coon M. J. Characterization of the omega-hydroxylase of Pseudomonas oleovorans as a nonheme iron protein. Arch Biochem Biophys. 1977 Oct;183(2):528–537. doi: 10.1016/0003-9861(77)90388-5. [DOI] [PubMed] [Google Scholar]
  13. Toms A., Wood J. M. Early intermediates in the degradation of alpha-conidendrin by a Pseudomonas multivorans. Biochemistry. 1970 Feb 17;9(4):733–740. doi: 10.1021/bi00806a005. [DOI] [PubMed] [Google Scholar]
  14. Tonge G. M., Harrison D. E., Knowles C. J., Higgins I. J. Properties and partial purification of the methane-oxidising enzyme system from Methylosinus trichosporium. FEBS Lett. 1975 Oct 15;58(1):293–299. doi: 10.1016/0014-5793(75)80282-1. [DOI] [PubMed] [Google Scholar]
  15. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  16. YPHANTIS D. A. EQUILIBRIUM ULTRACENTRIFUGATION OF DILUTE SOLUTIONS. Biochemistry. 1964 Mar;3:297–317. doi: 10.1021/bi00891a003. [DOI] [PubMed] [Google Scholar]

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