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Applied and Environmental Microbiology, October 2007, p. 6620-6628, Vol. 73, No. 20
0099-2240/07/$08.00+0     doi:10.1128/AEM.00881-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

The Ability of Aneurinibacillus migulanus (Bacillus brevis) To Produce the Antibiotic Gramicidin S Is Correlated with Phenotype Variation{triangledown}

Marina Berditsch,1 Sergii Afonin,2 and Anne S. Ulrich1,2*

Institute of Organic Chemistry, University of Karlsruhe, Fritz-Haber-Weg 6, 76131 Karlsruhe, Germany,1 Institute of Biological Interfaces, Forschungszentrum Karlsruhe, POB 3640, 76021 Karlsruhe, Germany2

Received 19 April 2007/ Accepted 14 August 2007

Phenotype instability of bacterial strains can cause significant problems in biotechnological applications, since industrially useful properties may be lost. Here we report such degenerative dissociation for Aneurinibacillus migulanus (formerly known as Bacillus brevis) an established producer of the antimicrobial peptide gramicidin S (GS). Phenotypic variations within and between various strains maintained in different culture collections are demonstrated. The type strain, ATCC 9999, consists of six colony morphology variants, R, RC, RP, RT, SC, and SP, which were isolated and characterized as pure cultures. Correlations between colony morphology, growth, GS production, spore formation, and resistance to their own antimicrobial peptide were established in this study. We found the original R form to be the best producer, followed by RC, RP, and RT, while SC and SP yielded no GS at all. Currently available ATCC 9999T contains only 2% of the original R producer and is dominated by the newly described phenotypes RC and RP. No original R form is detected in the nominally equivalent strain DSM 2895T (=ATCC 9999T), which grows only as SC and SP phenotypes and has thus completely lost its value as a peptide producer. Two other strains from the same collection, DSM 5668 and DSM 5759, contain the unproductive SC variant and the GS-producing RC form, respectively. We describe the growth and maintenance conditions that stabilize certain colony phenotypes and reduce the degree of degenerative dissociation, thus providing a recommendation for how to revert the nonproducing smooth phenotypes to the valuable GS-producing rough ones.


* Corresponding author. Mailing address: University of Karlsruhe (TH), Institute of Organic Chemistry, Fritz-Haber-Weg 6, 76131 Karlsruhe, Germany. Phone: 49(0721)6083912. Fax: 49(0721)6084823. E-mail: anne.ulrich{at}ibg.fzk.de

{triangledown} Published ahead of print on 24 August 2007.


Applied and Environmental Microbiology, October 2007, p. 6620-6628, Vol. 73, No. 20
0099-2240/07/$08.00+0     doi:10.1128/AEM.00881-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.




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