1887

Abstract

sp. strain PPD can metabolize phenanthrene as the sole source of carbon and energy via the ‘phthalic acid’ route. The key enzyme, 1-hydroxy-2-naphthoic acid dioxygenase (1-HNDO, EC 1.13.11.38), was purified to homogeneity using a 3-hydroxy-2-naphthoic acid (3-H2NA)-affinity matrix. The enzyme was a homotetramer with a native molecular mass of 160 kDa and subunit molecular mass of ∼39 kDa. It required Fe(II) as the cofactor and was specific for 1-hydroxy-2-naphthoic acid (1-H2NA), with 13.5 μM and 114 μmol min mg. 1-HNDO failed to show activity with gentisic acid, salicylic acid and other hydroxynaphthoic acids tested. Interestingly, the enzyme showed substrate inhibition with a of 116 μM. 1-HNDO was found to be competitively inhibited by 3-H2NA with a of 24 μM. Based on the pH-dependent spectral changes, the enzyme reaction product was identified as 2-carboxybenzalpyruvic acid. Under anaerobic conditions, the enzyme failed to convert 1-H2NA to 2-carboxybenzalpyruvic acid. Stoichiometric studies showed the incorporation of 1 mol O into the substrate to yield 1 mol product. These results suggest that 1-HNDO from sp. strain PPD is an extradiol-type ring-cleaving dioxygenase.

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2009-09-01
2020-08-08
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References

  1. Adachi K., Iwayama Y., Taniok H., Takeda Y.. 1966; Purification and properties of homogentisate oxygenase from Pseudomonas fluorescens. Biochim Biophys Acta118:88–97
    [Google Scholar]
  2. Adachi K., Iwabuchi T., Sano H., Harayama S.. 1999; Structure of the ring cleavage product of 1-hydroxy-2-naphthoate, an intermediate of the phenanthrene-degradative pathway of Nocardioides sp. strain KP7. J Bacteriol181:757–763
    [Google Scholar]
  3. Adams M. A., Singh V. K., Keller B. O., Jia Z.. 2006; Structural and biochemical characterization of gentisate 1,2-dioxygenase from Escherichia coli O157 : H7. Mol Microbiol61:1469–1484
    [Google Scholar]
  4. Arciero D. M., Lipscomb J. D.. 1986; Binding a 17O-labeled substrate and inhibitors to protocatechuate 4,5-dioxygenase nitrosyl complex: evidence for direct substrate binding to the active site Fe2+ of extradiol dioxygenases. J Biol Chem261:2170–2178
    [Google Scholar]
  5. Barnsley E. A.. 1983; Phthalate pathway of phenanthrene metabolism: formation of 2′-carboxybenzalpyruvate. J Bacteriol154:113–117
    [Google Scholar]
  6. Bradford M. M.. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem72:248–254
    [Google Scholar]
  7. Bucker M., Glatt H. R., Platt K. L., Avnir D., Ittah Y., Blum J., Oesch F.. 1979; Mutagenicity of phenanthrene and phenanthrene K-region derivatives. Mutat Res66:337–348
    [Google Scholar]
  8. Chang H. K., Zylstra G. J.. 1999; Characterization of the phthalate permease OphD from Burkholderia cepacia ATCC 17616. J Bacteriol181:6197–6199
    [Google Scholar]
  9. Crawford R. L., Hutton S. W., Chapman P. J.. 1975; Purification and properties of gentisate 1,2-dioxygenase from Moraxella osloensis. J Bacteriol121:794–799
    [Google Scholar]
  10. Davis M. I., Orville A. M., Neese F., Zaleski J. M., Lipscomb J. D., Solomon E. I.. 2002; Spectroscopic and electronic structure studies of protocatechuate 3,4-dioxygenase: nature of tyrosinate-Fe(III) bonds and their contribution to reactivity. J Am Chem Soc124:602–614
    [Google Scholar]
  11. Deveryshetty J., Suvekbala V., Varadamshetty G., Phale P. S.. 2007; Metabolism of 2-, 3- and 4-hydroxybenzoates by soil isolates Alcaligenes sp. strain PPH and Pseudomonas sp. strain PPD. FEMS Microbiol Lett268:59–66
    [Google Scholar]
  12. Doddamani H. P., Ninnekar H. Z.. 2000; Biodegradation of phenanthrene by a Bacillus species. Curr Microbiol41:11–14
    [Google Scholar]
  13. Evans W. C., Fernley H. N., Griffiths E.. 1965; Oxidative metabolism of phenanthrene and anthracene by soil pseudomonads. The ring-fission mechanism. Biochem J95:819–831
    [Google Scholar]
  14. Feng Y., Khoo H. E., Poh C. L.. 1999; Purification and characterization of gentisate 1,2-dioxygenases from Pseudomonas alcaligenes NCIB 9867 and Pseudomonas putida NCIB 9869. Appl Environ Microbiol65:946–950
    [Google Scholar]
  15. Ghosh D. K., Mishra A. K.. 1983; Oxidation of phenanthrene by a strain of Micrococcus: evidence of protocatechuate pathway. Curr Microbiol9:219–224
    [Google Scholar]
  16. Harpel M. R., Lipscomb J. D.. 1990; Gentisate 1,2-dioxygenase from Pseudomonas. Purification, characterization, and comparison of the enzymes from Pseudomonas testosteroni and Pseudomonas acidovorans. J Biol Chem265:6301–6311
    [Google Scholar]
  17. Hayaishi O., Hoshimoto K.. 1950; Pyrocatecase, a new enzyme catalyzing oxidative breakdown of pyrocatechin. J Biochem37:371–374
    [Google Scholar]
  18. Hintner J. P., Lechner C., Riegert U., Kuhm A. E., Storm T., Reemtsma T., Stolz A.. 2001; Direct ring fission of salicylate by a salicylate 1,2-dioxygenase activity from Pseudaminobacter salicylatoxidans. J Bacteriol183:6936–6942
    [Google Scholar]
  19. Hintner J. P., Reemtsma T., Stolz A.. 2004; Biochemical and molecular characterization of a ring fission dioxygenase with the ability to oxidize (substituted) salicylate(s) from Pseudaminobacter salicylatoxidans. J Biol Chem279:37250–37260
    [Google Scholar]
  20. Iwabuchi T., Harayama S.. 1997; Biochemical and genetic characterization of 2-carboxybenzaldehyde dehydrogenase, an enzyme involved in phenanthrene degradation by Nocardioides sp. strain KP7. J Bacteriol179:6488–6494
    [Google Scholar]
  21. Iwabuchi T., Harayama S.. 1998; Biochemical and molecular characterization of 1-hydroxy-2-naphthoate dioxygenase from Nocardioides sp. KP7. J Biol Chem273:8332–8336
    [Google Scholar]
  22. Kita A., Kita S., Fujisawa I., Inaka K., Ishida T., Horiike K., Nozaki M., Miki K.. 1999; An archetypical extradiol-cleaving catecholic dioxygenase: the crystal structure of catechol 2,3-dioxygenase (metapyrocatechase) from Pseudomonas putida mt-2. Structure7:25–34
    [Google Scholar]
  23. Kiyohara H., Nagao K., Nomi R.. 1976; Degradation of phenanthrene through o-phthalate by an Aeromonas sp. Agric Biol Chem40:1075–1082
    [Google Scholar]
  24. Kojima Y., Itada N., Hayaishi O.. 1961; Metapyrocatachase: a new catechol-cleaving enzyme. J Biol Chem236:2223–2228
    [Google Scholar]
  25. Laemmli U. K.. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature227:680–685
    [Google Scholar]
  26. March S. C., Parikh I., Cuatrecasas P.. 1974; A simplified method for cyanogen bromide activation of agarose for affinity chromatography. Anal Biochem60:149–152
    [Google Scholar]
  27. Mastrangelo G., Fadda E., Marzia V.. 1996; Polycyclic aromatic hydrocarbons and cancer in man. Environ Health Perspect104:1166–1170
    [Google Scholar]
  28. Matera I., Ferraroni M., Burger S., Scozzafava A., Stolz A., Briganti F.. 2008; Salicylate 1,2-dioxygenase from Pseudaminobacter salicylatoxidans: crystal structure of a peculiar ring-cleaving dioxygenase. J Mol Biol380:856–868
    [Google Scholar]
  29. Ono K., Nozaki M., Hayaishi O.. 1970; Purification and some properties of protocatechuate 4,5-dioxygenase. Biochim Biophys Acta220:224–238
    [Google Scholar]
  30. Phale P. S., Basu A., Majhi P. D., Deveryshetty J., Vamsee-Krishna C., Shrivastava R.. 2007; Metabolic diversity in bacterial degradation of aromatic compounds. OMICS11:252–279
    [Google Scholar]
  31. Prabhu Y., Phale P. S.. 2003; Biodegradation of phenanthrene by Pseudomonas sp. strain PP2: novel metabolic pathway, role of biosurfactant and cell surface hydrophobicity in hydrocarbon assimilation. Appl Microbiol Biotechnol61:342–351
    [Google Scholar]
  32. Samanta S. K., Chakraborti A. K., Jain R. K.. 1999; Degradation of phenanthrene by different bacteria: evidence for novel transformation sequences involving the formation of 1-naphthol. Appl Microbiol Biotechnol53:98–107
    [Google Scholar]
  33. Shu L., Chiou Y. M., Orville A. M., Miller M. A., Lipscomb J. D., Que L. Jr. 1995; X-ray absorption spectroscopic studies of the Fe(II) active site of catechol 2,3-dioxygenase. Implications for the extradiol cleavage mechanism. Biochemistry34:6649–6659
    [Google Scholar]
  34. Stanier R. Y., Ingraham J. L.. 1954; Protocatechuic acid oxidase. J Biol Chem210:799–808
    [Google Scholar]
  35. Sugumaran M., Vaidyanathan C. S.. 1978; Affinity chromatography of homogentisate 1,2-dioxygenase from Aspegillus niger. FEMS Microbiol Lett4:343–347
    [Google Scholar]
  36. Vamsee-Krishna C., Phale P. S.. 2008; Bacterial degradation of phthalate isomers and their esters. Indian J Microbiol48:19–34
    [Google Scholar]
  37. Vamsee-Krishna C., Mohan Y., Phale P. S.. 2006; Biodegradation of phthalate isomers by Pseudomonas aeruginosa PP4, Pseudomonas sp. PPD and Acinetobacter lwoffii ISP4. Appl Microbiol Biotechnol72:1263–1269
    [Google Scholar]
  38. Vetting M. W., Ohlendorf D. H.. 2000; The 1.8 Å crystal structure of catechol 1,2-dioxygenase reveals a novel hydrophobic helical zipper as a subunit linker. Structure8:429–440
    [Google Scholar]
  39. Vetting M. W., D'Argenio D. A., Ornston L. N., Ohlendorf D. H.. 2000; Structure of Acinetobacter strain ADP1 protocatechuate 3, 4-dioxygenase at 2.2 Å resolution: implications for the mechanism of an intradiol dioxygenase. Biochemistry39:7943–7955
    [Google Scholar]
  40. Werwath J., Arfmann H. A., Pieper D. H., Timmis K. N., Wittich R. M.. 1998; Biochemical and genetic characterization of a gentisate 1,2-dioxygenase from Sphingomonas sp. strain RW5. J Bacteriol180:4171–4176
    [Google Scholar]
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