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Volume 155, Issue 9

Biodegradation of phenanthrene by sp. strain PPD: purification and characterization of 1-hydroxy-2-naphthoic acid dioxygenase Free

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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Keyword(s): 1-H2NA, 1-hydroxy-2-naphthoic acid , 1-HNDO, 1-hydroxy-2-naphthoic acid dioxygenase , 2-CBADH, 2-carboxybenzaldehyde dehydrogenase , 2-H1NA, 2-hydroxy-1-naphthoic acid , 3-H2NA, 3-hydroxy-2-naphthoic acid , 6-H2NA, 6-hydroxy-2-naphthoic acid , GDO, gentisic acid 1,2-dioxygenase , P3,4DO, protocatechuate 3,4-dioxygenase , P4,5DO, protocatechuic acid 4,5-dioxygenase and SDO, salicylic acid 1,2-dioxygenase
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2009-09-01
2024-12-03
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Biodegradation of phenanthrene by Pseudomonas sp. strain PPD: purification and characterization of 1-hydroxy-2-naphthoic acid dioxygenase
Microbiology 155, 3083 (2009); https://doi.org/10.1099/mic.0.030460-0
/content/journal/micro/10.1099/mic.0.030460-0
/content/journal/micro/10.1099/mic.0.030460-0
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