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Antimicrobial Agents and Chemotherapy, February 2004, p. 514-520, Vol. 48, No. 2
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.2.514-520.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Streptomyces clavuligerus Has a Second Copy of the Proclavaminate Amidinohydrolase Gene

Susan E. Jensen,1* Annie Wong,1 Alison Griffin,2 and Barry Barton2

Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada T6G 2E9,1 Glaxo SmithKline Pharmaceuticals, Worthing, West Sussex BN14 8QH, England2

Received 29 April 2003/ Returned for modification 30 August 2003/ Accepted 19 October 2003

Past genetic studies have indicated that the genes encoding early enzymes of clavulanic acid biosynthesis may be duplicated in Streptomyces clavuligerus. We observed cross-hybridizing bands upon Southern analyses of proclavaminate amidinohydrolase (pah)-defective mutant strains of S. clavuligerus screened with a pah-specific probe. The DNA fragment responsible for this cross hybridization was cloned and sequenced and shown to encode a second copy of the pah gene. The new pah gene (pah1) was 1,056 bp in length, and its sequence was 72% identical to that of the original pah gene (pah2). Disruption mutants with defects in pah1 showed no significant effects on production of clavulanic acid or any of the clavam metabolites with stereochemistries opposite that of clavulanic acid (5S clavams) produced by S. clavuligerus when they were grown on starch asparagine or soy medium. However, double mutants with defects in both pah1 and pah2 were defective in the production of both clavulanic acid and all of the 5S clavam metabolites.

* Corresponding author. Mailing address: Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada T6G 2E9. Phone (780) 492-0830. Fax: (780) 492-9234. E-mail: susan.jensen{at}ualberta.ca.

Antimicrobial Agents and Chemotherapy, February 2004, p. 514-520, Vol. 48, No. 2
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.2.514-520.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Zelyas, N. J., Cai, H., Kwong, T., Jensen, S. E. (2008). Alanylclavam Biosynthetic Genes Are Clustered Together with One Group of Clavulanic Acid Biosynthetic Genes in Streptomyces clavuligerus. J. Bacteriol. 190: 7957-7965 [Abstract] [Full Text]  
  • Arulanantham, H., Kershaw, N. J., Hewitson, K. S., Hughes, C. E., Thirkettle, J. E., Schofield, C. J. (2006). ORF17 from the Clavulanic Acid Biosynthesis Gene Cluster Catalyzes the ATP-dependent Formation of N-Glycyl-clavaminic Acid. J. Biol. Chem. 281: 279-287 [Abstract] [Full Text]  
  • Bignell, D. R. D., Tahlan, K., Colvin, K. R., Jensen, S. E., Leskiw, B. K. (2005). Expression of ccaR, Encoding the Positive Activator of Cephamycin C and Clavulanic Acid Production in Streptomyces clavuligerus, Is Dependent on bldG. Antimicrob. Agents Chemother. 49: 1529-1541 [Abstract] [Full Text]  
  • Tahlan, K., Anders, C., Jensen, S. E. (2004). The Paralogous Pairs of Genes Involved in Clavulanic Acid and Clavam Metabolite Biosynthesis Are Differently Regulated in Streptomyces clavuligerus. J. Bacteriol. 186: 6286-6297 [Abstract] [Full Text]  


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