caa a22 4500 605506671 CHVBK 20210128100628.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s00253-015-6557-6 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6557-6 Engineering the heterologous expression of lanthipeptides in Escherichia coli by multigene assembly [Elektronische Daten] [Anja Kuthning, Eva Mösker, Roderich Süssmuth] Lantibiotics are an important class of ribosomally synthesised peptide antibiotics with a remarkable pharmacological potential. Structural variants of lantibiotics generated by peptide engineering in vivo are an important aspect for improving the peptide's efficacy, stability and bioavailability as well as production titre, which severely impacts the potential exploitation in pharmaceutical applications. Therefore, expression systems are needed which allow for a robust genetic access for ample mutagenesis experiments. Based on previous heterologous expression of the two-component lanthipeptide lichenicidin (Bliα and Bliβ) in Escherichia coli BLic5, we now employ a multigene assembly strategy for recombinant lantibiotic peptide production in the Gram-negative host. Two E. coli high copy plasmids for separate and increased expression of a two-component lantibiotic were cloned and tested for expression. From these E. coli HP expression strains, an up to 100 times increased expression was found compared with Bacillus licheniformis I89 and E. coli BLic5. Total expression yields reach 4mgL−1 for Bliα and 6mgL−1 for Bliβ. The expression system developed in this study constitutes an important cornerstone for future in vivo peptide engineering studies and is of significance for potential applications aiming at higher production titres of ribosomally synthesised, post translationally modified peptides. Springer-Verlag Berlin Heidelberg, 2015 Heterologous expression nationallicence Genetic engineering nationallicence Lantibiotic nationallicence Lichenicidin nationallicence Synthetic biology nationallicence Kuthning Anja Institut für Chemie, Technische Universität Berlin, Straße des 17 Juni 124, 10623, Berlin, Germany aut Mösker Eva Institut für Chemie, Technische Universität Berlin, Straße des 17 Juni 124, 10623, Berlin, Germany aut Süssmuth Roderich Institut für Chemie, Technische Universität Berlin, Straße des 17 Juni 124, 10623, Berlin, Germany aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/15(2015-08-01), 6351-6361 0175-7598 99:15<6351 2015 99 253 https://doi.org/10.1007/s00253-015-6557-6 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/s00253-015-6557-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kuthning Anja Institut für Chemie, Technische Universität Berlin, Straße des 17 Juni 124, 10623, Berlin, Germany aut NATIONALLICENCE 700 1- Mösker Eva Institut für Chemie, Technische Universität Berlin, Straße des 17 Juni 124, 10623, Berlin, Germany aut NATIONALLICENCE 700 1- Süssmuth Roderich Institut für Chemie, Technische Universität Berlin, Straße des 17 Juni 124, 10623, Berlin, Germany aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/15(2015-08-01), 6351-6361 0175-7598 99:15<6351 2015 99 253