caa a22 4500 605505101 CHVBK 20210128100620.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00253-014-6199-0 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6199-0 Bacillus subtilis as heterologous host for the secretory production of the non-ribosomal cyclodepsipeptide enniatin [Elektronische Daten] [Sophia Zobel, Jana Kumpfmüller, Roderich Süssmuth, Thomas Schweder] The heterologous expression of genes or gene clusters in microbial hosts, followed by metabolic engineering of biosynthetic pathways, is key to access industrially and pharmaceutically relevant compounds in an economically affordable and sustainable manner. Therefore, platforms need to be developed, which provide tools for the controlled synthesis of bioactive compounds. The Gram-positive bacterium Bacillus subtilis is a promising candidate for such applications, as it is generally regarded as a safe production host, its physiology is well investigated and a variety of tools is available for its genetic manipulation. Furthermore, this industrially relevant bacterium provides a high secretory potential not only for enzymes but also for primary and secondary metabolites. In this study, we present the first heterologous expression of an eukaryotic non-ribosomal peptide synthetase gene (esyn) coding for the biosynthesis of the small molecule enniatin in B. subtilis. Enniatin is a pharmaceutically used cyclodepsipeptide for treatment of topical bacterial and fungal infections. We generated various enniatin-producing B. subtilis strains, allowing for either single chromosomal or plasmid-based multi-copy expression of the esyn cluster under the control of an acetoin-inducible promoter system. Optimization of cultivation conditions, combined with modifications of the genetic background and multi-copy plasmid-based esyn expression, resulted in a secretory production of enniatin B. This work presents B. subtilis as a suitable host for the expression of heterologous eukaryotic non-ribosomal peptide synthetases (NRPS) clusters. The Author(s), 2014 Heterologous expression nationallicence Iterative NRPS nationallicence Cyclodepsipeptide nationallicence acoA promoter nationallicence Acetoin nationallicence Metabolic engineering nationallicence Zobel Sophia Institut für Chemie, Technische Universität Berlin, Strasse des 17. Juni 124, 10623, Berlin, Germany aut Kumpfmüller Jana Institut für Pharmazie, Ernst-Moritz-Arndt-Universität, Felix-Hausdorff-Strasse 3, 17489, Greifswald, Germany aut Süssmuth Roderich Institut für Chemie, Technische Universität Berlin, Strasse des 17. Juni 124, 10623, Berlin, Germany aut Schweder Thomas Institut für Pharmazie, Ernst-Moritz-Arndt-Universität, Felix-Hausdorff-Strasse 3, 17489, Greifswald, Germany aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/2(2015-01-01), 681-691 0175-7598 99:2<681 2015 99 253 https://doi.org/10.1007/s00253-014-6199-0 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6199-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zobel Sophia Institut für Chemie, Technische Universität Berlin, Strasse des 17. Juni 124, 10623, Berlin, Germany aut NATIONALLICENCE 700 1- Kumpfmüller Jana Institut für Pharmazie, Ernst-Moritz-Arndt-Universität, Felix-Hausdorff-Strasse 3, 17489, Greifswald, Germany aut NATIONALLICENCE 700 1- Süssmuth Roderich Institut für Chemie, Technische Universität Berlin, Strasse des 17. Juni 124, 10623, Berlin, Germany aut NATIONALLICENCE 700 1- Schweder Thomas Institut für Pharmazie, Ernst-Moritz-Arndt-Universität, Felix-Hausdorff-Strasse 3, 17489, Greifswald, Germany aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/2(2015-01-01), 681-691 0175-7598 99:2<681 2015 99 253