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Applied and Environmental Microbiology, January 2004, p. 224-228, Vol. 70, No. 1
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.1.224-228.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Novel Pathway for Utilization of Cyclopropanecarboxylate by Rhodococcus rhodochrous

Tetsuo Toraya,1* Takayuki Oka,1 Manabu Ando,1 Mamoru Yamanishi,1 and Hiroshi Nishihara2

Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University, Tsushima-Naka, Okayama 700-8530,1 Department of Chemistry, Faculty of Education, Kagawa University, Saiwai-cho, Takamatsu 760-8522, Japan2

Received 13 June 2003/ Accepted 23 September 2003

A new strain isolated from soil utilizes cyclopropanecarboxylate as the sole source of carbon and energy and was identified as Rhodococcus rhodochrous (H. Nishihara, Y. Ochi, H. Nakano, M. Ando, and T. Toraya, J. Ferment. Bioeng. 80:400-402, 1995). A novel pathway for the utilization of cyclopropanecarboxylate, a highly strained compound, by this bacterium was investigated. Cyclopropanecarboxylate-dependent reduction of NAD+ in cell extracts of cyclopropanecarboxylate-grown cells was observed. When intermediates accumulated in vitro in the absence of NAD+ were trapped as hydroxamic acids by reaction with hydroxylamine, cyclopropanecarboxohydroxamic acid and 3-hydroxybutyrohydroxamic acid were formed. Cyclopropanecarboxyl-coenzyme A (CoA), 3-hydroxybutyryl-CoA, and crotonyl-CoA were oxidized with NAD+ in cell extracts, whereas methacrylyl-CoA and 3-hydroxyisobutyryl-CoA were not. When both CoA and ATP were added, organic acids corresponding to the former three CoA thioesters were also oxidized in vitro by NAD+, while methacrylate, 3-hydroxyisobutyrate, and 2-hydroxybutyrate were not. Therefore, it was concluded that cyclopropanecarboxylate undergoes oxidative degradation through cyclopropanecarboxyl-CoA and 3-hydroxybutyryl-CoA. The enzymes catalyzing formation and ring opening of cyclopropanecarboxyl-CoA were shown to be inducible, while other enzymes involved in the degradation were constitutive.

* Corresponding author. Mailing address: Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University, Tsushima-Naka, Okayama 700-8530, Japan. Phone: 81-86-251-8194. Fax: 81-86-251-8264. E-mail: toraya{at}biotech.okayama-u.ac.jp.

Applied and Environmental Microbiology, January 2004, p. 224-228, Vol. 70, No. 1
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.1.224-228.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





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