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Use of green fluorescent protein and luciferase biomarkers to monitor survival and activity of Arthrobacter chlorophenolicus A6 cells during degradation of 4‐chlorophenol in soil
Author(s) -
Elväng Annelie M.,
Westerberg Karolina,
Jernberg Cecilia,
Jansson Janet K.
Publication year - 2001
Publication title -
environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.954
H-Index - 188
eISSN - 1462-2920
pISSN - 1462-2912
DOI - 10.1046/j.1462-2920.2001.00156.x
Subject(s) - arthrobacter , biology , luciferase , green fluorescent protein , bioremediation , population , microbiology and biotechnology , flow cytometry , microorganism , bacteria , bioreporter , chlorophenol , soil microbiology , food science , biochemistry , reporter gene , gene , chemistry , gene expression , transfection , genetics , demography , phenol , organic chemistry , sociology
The recently isolated novel species Arthrobacter chlorophenolicus A6 is capable of growth on and degradation of high concentrations of 4‐chlorophenol (up to 350 μg ml −1 ) as the sole carbon and energy source. This strain shows promise for bioremediation of environmental sites contaminated with high levels of chlorophenols. In this study, green fluorescent protein ( gfp ) or luciferase ( luc ) genes were used as biomarkers for monitoring cell number and activity, respectively, during degradation of 4‐chlorophenol by A. chlorophenolicus cells. The individual marked strains, Arthrobacter chlorophenolicus A6L ( luc ‐tagged) and Arthrobacter chlorophenolicus A6G ( gfp ‐tagged), were monitored during degradation of 250 μg ml −1 4‐chlorophenol in pure culture and 175 μg g −1 4‐chlorophenol in soil microcosms. Both gene‐tagged strains were capable of cleaning up the contaminated soil during 9 d incubation. During the bioremediation experiments, the luc ‐tagged cells were monitored using luminometry and the gfp ‐tagged cells using flow cytometry, in addition to selective plate counting for both strains. The cells remained at high population levels in the soil (evidenced by GFP‐fluorescent cell counts) and the A. chlorophenolicus A6L population was metabolically active (evidenced by luciferase activity measurements). These results demonstrate that the Arthrobacter chlorophenolicus A6 inoculum is effective for cleaning‐up soil containing high concentrations of 4‐chlorophenol.