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Applied and Environmental Microbiology, May 2004, p. 2830-2835, Vol. 70, No. 5
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.5.2830-2835.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Addition of Aromatic Substrates Restores Trichloroethylene Degradation Activity in Pseudomonas putida F1

Yuki Morono, Hajime Unno, Yasunori Tanji, and Katsutoshi Hori*

Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama 226-8501, Japan

Received 26 October 2003/ Accepted 27 January 2004

The rate of trichloroethylene (TCE) degradation by toluene dioxygenase (TDO) in resting cells of Pseudomonas putida F1 gradually decreased and eventually stopped within 1.5 h, as in previous reports. However, the subsequent addition of toluene, which is the principal substrate of TDO, resulted in its immediate degradation without a lag phase. After the consumption of toluene, degradation of TCE restarted at a rate similar to its initial degradation, suggesting that this degradation was mediated by TDO molecules that were present before the cessation of TCE degradation. The addition of benzene and cumene, which are also substrates of TDO, also caused restoration of TCE degradation activity: TCE was degraded simultaneously with cumene, and a larger amount of TCE was degraded after cumene was added than after toluene or benzene was added. But substrates that were expected to supply the cells with NADH or energy did not restore TCE degradation activity. This cycle of pseudoinactivation and restoration of TCE degradation was observed repeatedly without a significant decrease in the number of viable cells, even after six additions of toluene spread over 30 h. The results obtained in this study demonstrate a new type of restoration of TCE degradation that has not been previously reported.

* Corresponding author. Present address: Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan. Phone: 81-52-735-5214. Fax: 81-52-735-5255. E-mail: hori.katsutoshi{at}nitech.ac.jp.

Applied and Environmental Microbiology, May 2004, p. 2830-2835, Vol. 70, No. 5
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.5.2830-2835.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





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