Antibacterial Metabolites and Bacteriolytic Enzymes Produced by Bacillus pumilus During Bacteriolysis of Arthrobacter citreus
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The marine isolate Bacillus pumilus SBUG 1800 is able to lyse living cells of Arthrobacter citreus on solid media as well as pasteurized A. citreus cells in liquid mineral salt medium. The cultivation of B. pumilus in the presence of pasteurized A. citreus is accompanied by an enhanced production of 2,5-diketopiperazines (DKPs). DKPs inhibit bacterial growth, but do not seem to cause bacteriolysis. This study shows that B. pumilus also lyses living cells of A. citreus in co-culture experiments as an intraguild predator, even if the inoculum of B. pumilus is low. In order to characterize the bacteriolytic process, more precisely changes in the extracellular metabolome and proteome have been analyzed under different culture conditions. Besides the known DKPs, a number of different pumilacidins and bacteriolytic enzymes are produced. Two lipopeptides with [M + H]+ = 1008 and [M + H]+ = 1022 were detected and are proposed to be pumilacidin H and I. While the lipopeptides lyse living bacterial cells in lysis test assays, a set of extracellular enzymes degrades the dead cell material. Two of the cell wall hydrolases involved have been identified as N-acetylmuramoyl-l-alanine amidase and beta-N-acetylglucosaminidase. These findings together with electron microscopic and cell growth monitoring during co-culture experiments give a detailed view on the bacteriolytic process.
KeywordsN-Acetylmuramoyl-l-alanine amidase Beta-N-acetylglucosaminidase Diketopiperazines Pumilacidin I Pumilacidin H Bacterivory Predation
The authors thank Annette Meuche, Anne Reinhard, Stefan Bock, and Alexander Meene for laboratory assistance, Bob Jack for reviewing the manuscript, Dr. Birgit Voigt and Stefan Handtke for their helpful professional suggestions, and Dr. Dirk Albrecht for the MS/MS data analyzes of the native 1D gel and the 2D gels on which this work bases. We also thank the government of Mecklenburg-Vorpommern (Germany) for financial support in the form of a Landesgraduiertenstipendium.
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