Biochemical Characterization of Inducible 'Reductase' Component of Benzoate Dioxygenase and Phthalate Isomer Dioxygenases from Pseudomonas aeruginosa strain PP4
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Title |
Biochemical Characterization of Inducible 'Reductase' Component of Benzoate Dioxygenase and Phthalate Isomer Dioxygenases from Pseudomonas aeruginosa strain PP4
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Creator |
KARANDIKAR, R
BADRI, A PHALE, PS |
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Subject |
IRON-SULFUR FLAVOPROTEIN
ELECTRON-TRANSPORT PROTEINS TEREPHTHALATE 1,2-DIOXYGENASE NAPHTHALENE DIOXYGENASE OXYGENASE REDUCTASE PURIFICATION ENZYME CRYSTALLIZATION BIODEGRADATION MECHANISM Pseudomonas aeruginosa Phthalate isomer metabolism Phthalate isomer dioxygenase Reductase component Induction studies Kinetic properties |
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Description |
The first step involved in the degradation of phthalate isomers (phthalate, isophthalate and terephthalate) is the double hydroxylation by respective aromatic-ring hydroxylating dioxygenases. These are two component enzymes consisting of 'oxygenase' and 'reductase' components. Soil isolate Pseudomonas aeruginosa strain PP4 degrades phthalate isomers via protocatechuate and benzoate via catechol 'ortho' ring cleavage pathway. Metabolic studies suggest that strain PP4 has carbon source-specific inducible phthalate isomer dioxygenase and benzoate dioxygenase. Thus, it was of interest to study the properties of reductase components of these enzymes. Reductase activity from phthalate isomer-grown cells was 3-5-folds higher than benzoate grown cells. In-gel activity staining profile showed a reductase activity band of R (f) 0.56 for phthalate isomer-grown cells as compared to R (f) 0.73 from benzoate-grown cells. Partially purified reductase components from phthalate isomer grown cells showed K (m) in the range of 30-40 mu M and V (max) = 34-48 mu mol min(-1) mg(-1). However, reductase from benzoate grown cells showed K (m) = 49 mu M and V (max) = 10 mu mol min(-1) mg(-1). Strikingly similar molecular and kinetic properties of reductase component from phthalate isomer-grown cells suggest that probably the same reductase component is employed in three phthalate isomer dioxygenases. However, reductase component is different, with respect to kinetic properties and zymogram analysis, from benzoate-grown cells when compared to that from phthalate isomer grown cells of PP4.
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Publisher |
HUMANA PRESS INC
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Date |
2016-01-14T12:29:42Z
2016-01-14T12:29:42Z 2015 |
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Type |
Article
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Identifier |
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY, 177(2)318-333
0273-2289 1559-0291 http://dx.doi.org/10.1007/s12010-015-1744-6 http://dspace.library.iitb.ac.in/jspui/handle/100/17500 |
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Language |
en
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