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Applied and Environmental Microbiology, March 2004, p. 1385-1392, Vol. 70, No. 3
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.3.1385-1392.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Aerobic and Anaerobic Toluene Degradation by a Newly Isolated Denitrifying Bacterium, Thauera sp. Strain DNT-1

Yoshifumi Shinoda,1 Yasuyoshi Sakai,1 Hiroshi Uenishi,1 Yasumitsu Uchihashi,1 Akira Hiraishi,2 Hideaki Yukawa,3 Hiroya Yurimoto,1 and Nobuo Kato1*

Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502,1 Department of Ecological Engineering, Toyohashi University of Technology, Toyohashi 441-8580,2 Research Institute of Innovative Technology for the Earth, Soraku-gun, Kyoto 619-0292, Japan3

Received 16 September 2003/ Accepted 25 November 2003

A newly isolated denitrifying bacterium, Thauera sp. strain DNT-1, grew on toluene as the sole carbon and energy source under both aerobic and anaerobic conditions. When this strain was cultivated under oxygen-limiting conditions with nitrate, first toluene was degraded as oxygen was consumed, while later toluene was degraded as nitrate was reduced. Biochemical observations indicated that initial degradation of toluene occurred through a dioxygenase-mediated pathway and the benzylsuccinate pathway under aerobic and denitrifying conditions, respectively. Homologous genes for toluene dioxygenase (tod) and benzylsuccinate synthase (bss), which are the key enzymes in aerobic and anaerobic toluene degradation, respectively, were cloned from genomic DNA of strain DNT-1. The results of Northern blot analyses and real-time quantitative reverse transcriptase PCR suggested that transcription of both sets of genes was induced by toluene. In addition, the tod genes were induced under aerobic conditions, whereas the bss genes were induced under both aerobic and anaerobic conditions. On the basis of these results, it is concluded that strain DNT-1 modulates the expression of two different initial pathways of toluene degradation according to the availability of oxygen in the environment.

* Corresponding author. Mailing address: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa, Sakyo-ku, Kyoto 606-8502, Japan. Phone: 81-75-753-6385. Fax: 81-75-753-6385. E-mail: nkato{at}kais.kyoto-u.ac.jp.

Applied and Environmental Microbiology, March 2004, p. 1385-1392, Vol. 70, No. 3
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.3.1385-1392.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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