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Journal of Bacteriology, December 1998, p. 6571-6580, Vol. 180, No. 24
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Cytochrome bd Biosynthesis in Bacillus subtilis: Characterization of the cydABCD Operon

Lena Winstedt,1 Ken-Ichi Yoshida,2 Yasutaro Fujita,2 and Claes von Wachenfeldt1,*

Department of Microbiology, Lund University, Lund, Sweden,1 and Department of Biotechnology, Faculty of Engineering, Fukuyama University, Fukuyama, Japan2

Received 26 June 1998/Accepted 14 October 1998

Under aerobic conditions Bacillus subtilis utilizes a branched electron transport chain comprising various cytochromes and terminal oxidases. At present there is evidence for three types of terminal oxidases in B. subtilis: a caa3-, an aa3-, and a bd-type oxidase. We report here the cloning of the structural genes (cydA and cydB) encoding the cytochrome bd complex. Downstream of the structural genes, cydC and cydD are located. These genes encode proteins showing similarity to bacterial ATP-binding cassette (ABC)-type transporters. Analysis of isolated cell membranes showed that inactivation of cydA or deletion of cydABCD resulted in the loss of spectral features associated with cytochrome bd. Gene disruption experiments and complementation analysis showed that the cydC and cydD gene products are required for the expression of a functional cytochrome bd complex. Disruption of the cyd genes had no apparent effect on the growth of cells in broth or defined media. The expression of the cydABCD operon was investigated by Northern blot analysis and by transcriptional and translational cyd-lacZ fusions. Northern blot analysis confirmed that cydABCD is transcribed as a polycistronic message. The operon was found to be expressed maximally under conditions of low oxygen tension.

* Corresponding author. Mailing address: Lund University, Department of Microbiology, Sölvegatan 12, S-223 62 Lund, Sweden. Phone: 46 46 2223456. Fax: 46 46 157839. E-mail: claes_von.wachenfeldt{at}mikrbiol.lu.se.

Journal of Bacteriology, December 1998, p. 6571-6580, Vol. 180, No. 24
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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