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Appl. Environ. Microbiol., Mar 1997, 1189-1191, Vol 63, No. 3
Copyright © 1997, American Society for Microbiology

Use of 4-Nitrophenoxyacetic Acid for Detection and Quantification of 2,4-Dichlorophenoxyacetic Acid (2,4-D)/(alpha)-Ketoglutarate Dioxygenase Activity in 2,4-D-Degrading Microorganisms

TM Sassanella, F Fukumori, M Bagdasarian and RP Hausinger
Department of Microbiology, Genetics Program, Department of Biochemistry, and NSF Center for Microbial Ecology, Michigan State University, East Lansing, Michigan 48824, and Research and Development Corporation of Japan, Kawaguchi, Saitama, Tokyo 100, Japan

Purified 2,4-dichlorophenoxyacetic acid (2,4-D)/(alpha)-ketoglutarate dioxygenase (TfdA) was shown to use 4-nitrophenoxyacetic acid (K(infm) = 0.89 (plusmn) 0.04 mM, k(infcat) [catalytic constant] = 540 (plusmn) 10 min(sup-1)), producing intensely yellow 4-nitrophenol. This reagent was used to develop a rapid, continuous, colorimetric assay for the detection of TfdA and analogous activities in 2,4-D-degrading bacterial cells and extracts.