Please use this identifier to cite or link to this item: http://dspace.library.iitb.ac.in/xmlui/handle/10054/11995
Title: Biodegradation of phenanthrene by Pseudomonas sp strain PPD: purification and characterization of 1-hydroxy-2-naphthoic acid dioxygenase
Authors: DEVERYSHETTY, J
PHALE, PS
Keywords: Gentisate 1,2-Dioxygenase
Crystal-Structure
Pseudaminobacter-Salicylatoxidans
Protocatechuate 3,4-Dioxygenase
Molecular Characterization
Catechol 2,3-Dioxygenase
Genetic-Characterization
Affinity Chromatography
Bacterial-Degradation
Phthalate Isomers
Issue Date: 2009
Publisher: SOC GENERAL MICROBIOLOGY
Citation: MICROBIOLOGY-SGM, 155(), 3083-3091
Abstract: Pseudomonas sp. strain PPID 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 similar to 39 kDa. It required Fe(II) as the cofactor and was specific for 1-hydroxy-2-naphthoic acid (1-H2NA), with K(m) 13.5 mu M and V(max) 114 mu mol min(-1) mg(-1). 1-HNDO failed to show activity with gentisic acid, salicylic acid and other hydroxynaphthoic acids tested. Interestingly, the enzyme showed substrate inhibition with a K(i) of 116 mu M. 1-HNDO was found to be competitively inhibited by 3-H2NA with a K(i) of 24 mu M. 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 02 into the substrate to yield 1 mol product. These results suggest that 1-HNDO from Pseudomonas sp. strain PPD is an extradiol-type ring-cleaving dioxygenase.
URI: http://dx.doi.org/10.1099/mic.0.030460-0
http://dspace.library.iitb.ac.in/xmlui/handle/10054/11995
http://hdl.handle.net/10054/11995
ISSN: 1350-0872
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