Chemical quantification of N-acyl alanine methyl ester (NAME) production and impact on temporal gene expression patterns in Roseovarius tolerans EL-164
Abstract Background Previous studies have identified structurally diverse N-acyl amino acid methyl esters (NAMEs) in culture extracts of Roseovarius tolerans EL-164 (Roseobacteraceae). NAMEs are structural analogues of the common signaling compounds N-acyl homoserine lactones (AHLs), but do not part...
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2024-11-01
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| Online Access: | https://doi.org/10.1186/s12866-024-03624-7 |
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| author | Janina Leinberger Diana Koteska Judith Boldt Jörn Petersen Sahana Shivaramu Jürgen Tomasch Stefan Schulz Thorsten Brinkhoff |
| author_facet | Janina Leinberger Diana Koteska Judith Boldt Jörn Petersen Sahana Shivaramu Jürgen Tomasch Stefan Schulz Thorsten Brinkhoff |
| author_sort | Janina Leinberger |
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| description | Abstract Background Previous studies have identified structurally diverse N-acyl amino acid methyl esters (NAMEs) in culture extracts of Roseovarius tolerans EL-164 (Roseobacteraceae). NAMEs are structural analogues of the common signaling compounds N-acyl homoserine lactones (AHLs), but do not participate in AHL-mediated signaling. NAMEs show minor antialgal and antimicrobial activity, but whether this activity serves as the primary ecological role remains unclear. Results To enable dose-dependent bioactivity-testing, we have established a chromatographic method for quantification of NAMEs in bacterial culture extracts. The concentrations determined for the two major NAMEs produced by EL-164, C16:1-NAME and C17:1-NAME, ranged between 0.685 and 5.731 mg L− 1 (2.0-16.9 µM) and 5.3–86.4 µg L− 1 (15.0-244.3 nM), respectively. Co-quantification of the C14:1-AHL showed concentrations ranging between 17.5 and 58.7 mg L− 1 (56.6-189.7 µM). We observed distinct production patterns for NAMEs and AHLs, with a continuous NAME production during the entire incubation period. We conducted a spike-in experiment, using the determined metabolite concentrations. By comparing the transcriptomes of pre- and post-metabolite-spikes, we identified three clusters of differentially expressed genes with distinct temporal expression patterns. Expression levels of stress response genes differed between NAME- and AHL-spiked EL-164 cultures in the stationary phase. Conclusions Our findings support previous studies suggesting an ecological role for C16:1-NAME as antibiotic, by proving that NAME concentrations in batch cultures were higher than the minimal inhibitory concentrations against Maribacter sp. 62 − 1 (Flavobacteriia) and Skeletonema costatum CCMP 1332 (Coscinodiscophyceae) reported in the literature. Our study further exemplified the broad application range of dose-dependent testing and highlighted the different biological activities of NAMEs and AHLs. |
| format | Article |
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| institution | OA Journals |
| issn | 1471-2180 |
| language | English |
| publishDate | 2024-11-01 |
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| series | BMC Microbiology |
| spelling | doaj-art-49a554fb8f73451091cf97ead18f55382025-08-20T02:32:56ZengBMCBMC Microbiology1471-21802024-11-0124111310.1186/s12866-024-03624-7Chemical quantification of N-acyl alanine methyl ester (NAME) production and impact on temporal gene expression patterns in Roseovarius tolerans EL-164Janina Leinberger0Diana Koteska1Judith Boldt2Jörn Petersen3Sahana Shivaramu4Jürgen Tomasch5Stefan Schulz6Thorsten Brinkhoff7Institute for Chemistry and Biology of the Marine Environment, University of OldenburgInstitute of Organic Chemistry, Technische Universität BraunschweigLeibniz Institute DSMZ - German Collection of Microorganisms and Cell CulturesLeibniz Institute DSMZ - German Collection of Microorganisms and Cell CulturesLaboratory of Anoxygenic Phototrophs, Institute of Microbiology of the Czech Acad SciLaboratory of Anoxygenic Phototrophs, Institute of Microbiology of the Czech Acad SciInstitute of Organic Chemistry, Technische Universität BraunschweigInstitute for Chemistry and Biology of the Marine Environment, University of OldenburgAbstract Background Previous studies have identified structurally diverse N-acyl amino acid methyl esters (NAMEs) in culture extracts of Roseovarius tolerans EL-164 (Roseobacteraceae). NAMEs are structural analogues of the common signaling compounds N-acyl homoserine lactones (AHLs), but do not participate in AHL-mediated signaling. NAMEs show minor antialgal and antimicrobial activity, but whether this activity serves as the primary ecological role remains unclear. Results To enable dose-dependent bioactivity-testing, we have established a chromatographic method for quantification of NAMEs in bacterial culture extracts. The concentrations determined for the two major NAMEs produced by EL-164, C16:1-NAME and C17:1-NAME, ranged between 0.685 and 5.731 mg L− 1 (2.0-16.9 µM) and 5.3–86.4 µg L− 1 (15.0-244.3 nM), respectively. Co-quantification of the C14:1-AHL showed concentrations ranging between 17.5 and 58.7 mg L− 1 (56.6-189.7 µM). We observed distinct production patterns for NAMEs and AHLs, with a continuous NAME production during the entire incubation period. We conducted a spike-in experiment, using the determined metabolite concentrations. By comparing the transcriptomes of pre- and post-metabolite-spikes, we identified three clusters of differentially expressed genes with distinct temporal expression patterns. Expression levels of stress response genes differed between NAME- and AHL-spiked EL-164 cultures in the stationary phase. Conclusions Our findings support previous studies suggesting an ecological role for C16:1-NAME as antibiotic, by proving that NAME concentrations in batch cultures were higher than the minimal inhibitory concentrations against Maribacter sp. 62 − 1 (Flavobacteriia) and Skeletonema costatum CCMP 1332 (Coscinodiscophyceae) reported in the literature. Our study further exemplified the broad application range of dose-dependent testing and highlighted the different biological activities of NAMEs and AHLs.https://doi.org/10.1186/s12866-024-03624-7Secondary metabolitesSignalingAntibiosisRoseobacterGrowth phasesQuantification |
| spellingShingle | Janina Leinberger Diana Koteska Judith Boldt Jörn Petersen Sahana Shivaramu Jürgen Tomasch Stefan Schulz Thorsten Brinkhoff Chemical quantification of N-acyl alanine methyl ester (NAME) production and impact on temporal gene expression patterns in Roseovarius tolerans EL-164 BMC Microbiology Secondary metabolites Signaling Antibiosis Roseobacter Growth phases Quantification |
| title | Chemical quantification of N-acyl alanine methyl ester (NAME) production and impact on temporal gene expression patterns in Roseovarius tolerans EL-164 |
| title_full | Chemical quantification of N-acyl alanine methyl ester (NAME) production and impact on temporal gene expression patterns in Roseovarius tolerans EL-164 |
| title_fullStr | Chemical quantification of N-acyl alanine methyl ester (NAME) production and impact on temporal gene expression patterns in Roseovarius tolerans EL-164 |
| title_full_unstemmed | Chemical quantification of N-acyl alanine methyl ester (NAME) production and impact on temporal gene expression patterns in Roseovarius tolerans EL-164 |
| title_short | Chemical quantification of N-acyl alanine methyl ester (NAME) production and impact on temporal gene expression patterns in Roseovarius tolerans EL-164 |
| title_sort | chemical quantification of n acyl alanine methyl ester name production and impact on temporal gene expression patterns in roseovarius tolerans el 164 |
| topic | Secondary metabolites Signaling Antibiosis Roseobacter Growth phases Quantification |
| url | https://doi.org/10.1186/s12866-024-03624-7 |
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