Taxonomic identification, phenanthrene uptake activity, and membrane lipid alterations of the PAH degrading Arthrobacter sp. strain Sphe3

Abstract

This report describes phenanthrene uptake as well as the effect of phenanthrene on the membrane phospholipid and fatty acid composition in a newly isolated bacterial strain, Sphe3, that we taxonomically identified as Arthrobacter sp. Strain Sphe3 is able to utilize phenanthrene as a carbon source at high rates and appears to internalize phenanthrene with two mechanisms: a passive diffusion when cells are grown on glucose, and an inducible active transport system when cells are grown on phenanthrene as a sole carbon source. Active transport followed Michaelis-Menten kinetics, and it was amenable to inhibition by 2,4-dinitrophenol and sodium azide. Evidence provided here indicates that apart from inducing an active PAH uptake, the presence of phenanthrene elicits significant changes in membrane fluidity.

Arthrobacter phenanthrenivorans sp. nov., to accommodate the phenanthrene-degrading bacterium Arthrobacter sp. strain Sphe3

“…We have previously reported the phenanthrene uptake activity and membrane lipid alterations of a PAH-degrading Arthrobacter strain, Sphe3, isolated from a creosote-contaminated soil in Greece (Kallimanis et al, 2007). In the present study, on the basis of phylogenetic analysis of 16S rRNA and recA gene sequences, together with physiological and chemotaxonomic characteristics and DNA-DNA hybridization analysis, we demonstrate that strain Sphe3 represents a novel species of the genus Arthrobacter.Strain Sphe3 T was isolated from Perivleptos, a creosotepolluted site in Epirus, Greece (12 km north of the city of Ioannina), in which a wood-preserving industry had operated for over 30 years (Kallimanis et al, 2007). Soil samples (10 g) were mixed with 100 ml of minimal medium M9 (Sambrook et al, 1989) supplemented with 0.01 % (w/v) phenanthrene (in crystal form) as the sole carbon and energy source for growth.…”

“…Among these micro-organisms, several Arthrobacter species are able to degrade PAHs (Grifoll et al, 1992;Seo et al, 2006). We have previously reported the phenanthrene uptake activity and membrane lipid alterations of a PAH-degrading Arthrobacter strain, Sphe3, isolated from a creosote-contaminated soil in Greece (Kallimanis et al, 2007). In the present study, on the basis of phylogenetic analysis of 16S rRNA and recA gene sequences, together with physiological and chemotaxonomic characteristics and DNA-DNA hybridization analysis, we demonstrate that strain Sphe3 represents a novel species of the genus Arthrobacter.…”

“…Phospholipid and fatty acid methyl ester analyses were performed as described previously (Kallimanis et al, 2007, and references therein). Isoprenoid quinone extraction was performed as described by Rosa-Putra et al (2001) and isoprenoid quinones were characterized by electrospray ionization mass spectrometry (ESI-MS).…”

“…Strain Sphe3 T was isolated from Perivleptos, a creosotepolluted site in Epirus, Greece (12 km north of the city of Ioannina), in which a wood-preserving industry had operated for over 30 years (Kallimanis et al, 2007). Soil samples (10 g) were mixed with 100 ml of minimal medium M9 (Sambrook et al, 1989) supplemented with 0.01 % (w/v) phenanthrene (in crystal form) as the sole carbon and energy source for growth.…”

Heterologous Expression and Characterization of Two 1-Hydroxy-2-Naphthoic Acid Dioxygenases from Arthrobacter phenanthrenivorans

“…We have previously reported the isolation of Arthrobacter phenanthrenivorans strain Sphe3, which can grow on phenanthrene as the sole source of carbon and energy, efficiently catabolizing phenanthrene up to 400 mg · liter Ϫ1 at high rates (22). In the present work, we describe the purification, catalytic properties, cloning, and characterization of two 1-H2NA dioxygenases isolated from A. phenanthrenivorans sp.…”