A novel degradative pathway of 2-nitrobenzoate via 3-hydroxyanthranilate in Pseudomonas fluorescens strain KU-7.
A bacterial strain KU-7, identified as a Pseudomonas fluorescens by 16S rDNA sequencing, was one of the 12 new isolates that are able to grow on 2-nitrobenzoate as a sole source of carbon, nitrogen, and energy. Resting cells of KU-7 were found to accumulate ammonia in the medium indicating that degradation of 2-NBA proceeds through a reductive route. Metabolite analyses by thin layer chromatography and high pressure liquid chromatography indicated that 3-hydroxyanthranilate is an intermediate of 2-nitrobenzoate metabolism in KU-7 cells. This offers an alternative route to 2-nitrobenzoate metabolism since anthranilate (2-aminobenzoate) or catechol were detected as intermediates in other bacteria. Crude extracts of KU-7 cells converted 2-nitrobenzoate to 3-hydroxyanthranilate with oxidation of 2 mol of NADPH. Ring cleavage of 3-hydroxyanthranilate produced a transient yellow product, identified as 2-amino-3-carboxymuconic 6-semialdehyde, that has a maximum absorbance at 360 nm. The initial enzymes of the 2-nitrobenzoate degradation pathway were found to be inducible since succinate-grown cells produced very low enzyme activities. A pathway for 2-nitrobenzoate degradation in KU-7 was proposed.
Hasegawa
Y
Y
Department of Biotechnology, Faculty of Engineering and High Technology Research Center, Kansai University, Osaka, Japan. yoshie@ipcku.kansai-u.ac.jp
Muraki
T
T
Tokuyama
T
T
Iwaki
H
H
Tatsuno
M
M
Lau
P C
PC
D015095:3-Hydroxyanthranilic Acid / Q000378:metabolism*; D001673:Biodegradation, Environmental; D003470:Culture Media; D009579:Nitrobenzoates / Q000378:metabolism*; D010578:Petroleum; D011551:Pseudomonas fluorescens / Q000254:growth & development / Q000302:isolation & purification / Q000378:metabolism*; D012988:Soil Microbiology; D014871:Water Microbiology; D014876:Water Pollution
D003470:Culture Media; D009579:Nitrobenzoates; D010578:Petroleum; D015095:3-Hydroxyanthranilic Acid