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| Natura: | Artículo científico |
| Lingua: | en |
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BMC microbiology
2024
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| Accesso online: | https://pubmed.ncbi.nlm.nih.gov/39574024/ |
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| author | Leinberger, Janina Koteska, Diana Boldt, Judith Petersen, Jörn Shivaramu, Sahana Tomasch, Jürgen Schulz, Stefan Brinkhoff, Thorsten |
| author_facet | Leinberger, Janina Koteska, Diana Boldt, Judith Petersen, Jörn Shivaramu, Sahana Tomasch, Jürgen Schulz, Stefan Brinkhoff, Thorsten Leinberger, Janina Koteska, Diana Boldt, Judith Petersen, Jörn Shivaramu, Sahana Tomasch, Jürgen Schulz, Stefan Brinkhoff, Thorsten |
| collection | PubMed - marine biology |
| contents | Chemical quantification of N-acyl alanine methyl ester (NAME) production and impact on temporal gene expression patterns in Roseovarius tolerans EL-164. Leinberger, Janina Koteska, Diana Boldt, Judith Petersen, Jörn Shivaramu, Sahana Tomasch, Jürgen Schulz, Stefan Brinkhoff, Thorsten Gene Expression Regulation, Bacterial Rhodobacteraceae Alanine Acyl-Butyrolactones Esters 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. 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 (2.0-16.9 µM) and 5.3-86.4 µg L (15.0-244.3 nM), respectively. Co-quantification of the C14:1-AHL showed concentrations ranging between 17.5 and 58.7 mg L (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. 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 | Artículo científico |
| id | pubmed_39574024 |
| institution | PubMed |
| language | en |
| publishDate | 2024 |
| publisher | BMC microbiology |
| record_format | pubmed |
| spellingShingle | Chemical quantification of N-acyl alanine methyl ester (NAME) production and impact on temporal gene expression patterns in Roseovarius tolerans EL-164. Leinberger, Janina Koteska, Diana Boldt, Judith Petersen, Jörn Shivaramu, Sahana Tomasch, Jürgen Schulz, Stefan Brinkhoff, Thorsten Gene Expression Regulation, Bacterial Rhodobacteraceae Alanine Acyl-Butyrolactones Esters Chemical quantification of N-acyl alanine methyl ester (NAME) production and impact on temporal gene expression patterns in Roseovarius tolerans EL-164. Leinberger, Janina Koteska, Diana Boldt, Judith Petersen, Jörn Shivaramu, Sahana Tomasch, Jürgen Schulz, Stefan Brinkhoff, Thorsten Gene Expression Regulation, Bacterial Rhodobacteraceae Alanine Acyl-Butyrolactones Esters 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. 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 (2.0-16.9 µM) and 5.3-86.4 µg L (15.0-244.3 nM), respectively. Co-quantification of the C14:1-AHL showed concentrations ranging between 17.5 and 58.7 mg L (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. 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. |
| title | Chemical quantification of N-acyl alanine methyl ester (NAME) production and impact on temporal gene expression patterns in Roseovarius tolerans EL-164. |
| topic | Gene Expression Regulation, Bacterial Rhodobacteraceae Alanine Acyl-Butyrolactones Esters |
| url | https://pubmed.ncbi.nlm.nih.gov/39574024/ |