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Applied and Environmental Microbiology, February 2002, p. 668-672, Vol. 68, No. 2
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.2.668-672.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Characterization of the norB Gene, Encoding Nitric Oxide Reductase, in the Nondenitrifying Cyanobacterium Synechocystis sp. Strain PCC6803

Andrea Büsch, Bärbel Friedrich, and Rainer Cramm*

Institut für Biologie/Mikrobiologie, Humboldt-Universität zu Berlin, D-10115 Berlin, Germany

Received 7 August 2001/ Accepted 14 November 2001

A norB gene encoding a putative nitric oxide reductase is present in the genome of the nondenitrifying cyanobacterium Synechocystis sp. strain PCC6803. The gene product belongs to the quinol-oxidizing single-subunit class of nitric oxide reductases, discovered recently in the denitrifier Ralstonia eutropha. Heterologous complementation of a nitric oxide reductase-negative mutant of R. eutropha with norB from Synechocystis restored nitric oxide reductase activity. With reduced menadione as the electron donor, an enzymatic activity of 101 nmol of NO per min per mg of protein was obtained with membrane fractions of Synechocystis wild-type cells. Virtually no nitric oxide reductase activity was present in a norB-negative mutant of Synechocystis. Growing cells of this mutant are more sensitive toward NO than wild-type cells, indicating that the presence of a nitric oxide reductase is beneficial for Synechocystis when the cells are exposed to NO. Transcriptional fusions with the chloramphenicol acetyltransferase reporter gene were constructed to monitor norB expression in Synechocystis. Transcription of norB was not enhanced by the addition of the NO-generating agent sodium nitroprusside.

* Corresponding author. Mailing address: Institut für Biologie/Mikrobiologie, Humboldt-Universität zu Berlin, Chausseestr. 117, D-10115 Berlin, Germany. Phone: 49-30-20938108. Fax: 49-30-20938102. E-mail: rainer.cramm{at}rz.hu-berlin.de.

Applied and Environmental Microbiology, February 2002, p. 668-672, Vol. 68, No. 2
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.2.668-672.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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