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Journal of Bacteriology, May 2001, p. 3184-3192, Vol. 183, No. 10
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.10.3184-3192.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Role of Acid Metabolism in Streptomyces coelicolor Morphological Differentiation and Antibiotic Biosynthesis

Patrick H. Viollier,1,dagger Wolfgang Minas,2 Glenn E. Dale,1,Dagger Marc Folcher,1 and Charles J. Thompson1,*

Department of Molecular Microbiology, Biozentrum, University of Basel, Basel,1 and Institute of Biotechnology, ETH-Zürich, HPT, Zürich,2 Switzerland

Received 6 September 2000/Accepted 8 February 2001

Studies of citrate synthase (CitA) were carried out to investigate its role in morphological development and biosynthesis of antibiotics in Streptomyces coelicolor. Purification of CitA, the major vegetative enzyme activity, allowed characterization of its kinetic properties. The apparent Km values of CitA for acetyl coenzyme A (acetyl-CoA) (32 µM) and oxaloacetate (17 µM) were similar to those of citrate synthases from other gram-positive bacteria and eukaryotes. CitA was not strongly inhibited by various allosteric feedback inhibitors (NAD+, NADH, ATP, ADP, isocitrate, or alpha -ketoglutarate). The corresponding gene (citA) was cloned and sequenced, allowing construction of a citA mutant (BZ2). BZ2 was a glutamate auxotroph, indicating that citA encoded the major citrate synthase allowing flow of acetyl-CoA into the tricarboxylic acid (TCA) cycle. Interruption of aerobic TCA cycle-based metabolism resulted in acidification of the medium and defects in morphological differentiation and antibiotic biosynthesis. These developmental defects of the citA mutant were in part due to a glucose-dependent medium acidification that was also exhibited by some other bald mutants. Unlike other acidogenic bald strains, citA and bldJ mutants were able to produce aerial mycelia and pigments when the medium was buffered sufficiently to maintain neutrality. Extracellular complementation studies suggested that citA defines a new stage of the Streptomyces developmental cascade.

* Corresponding author. Mailing address: Biozentrum, University of Basel, Department of Molecular Microbiology, Klingelbergstrasse 70, CH-4056 Basel, Switzerland. Phone: 41 61 267 2116. Fax: 41 62 267 2118. E-mail: charles-j.thompson{at}unibas.ch.

dagger Present address: Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA 94305-5329.

Dagger Present address: Morphochem, Basel, BS 4058, Switzerland.

Journal of Bacteriology, May 2001, p. 3184-3192, Vol. 183, No. 10
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.10.3184-3192.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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