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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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