Engineering the heterologous expression of lanthipeptides in Escherichia coli by multigene assembly

Verfasser / Beitragende:
[Anja Kuthning, Eva Mösker, Roderich Süssmuth]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/15(2015-08-01), 6351-6361
Format:
Artikel (online)
ID: 605506671
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024 7 0 |a 10.1007/s00253-015-6557-6  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-015-6557-6 
245 0 0 |a Engineering the heterologous expression of lanthipeptides in Escherichia coli by multigene assembly  |h [Elektronische Daten]  |c [Anja Kuthning, Eva Mösker, Roderich Süssmuth] 
520 3 |a 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. 
540 |a Springer-Verlag Berlin Heidelberg, 2015 
690 7 |a Heterologous expression  |2 nationallicence 
690 7 |a Genetic engineering  |2 nationallicence 
690 7 |a Lantibiotic  |2 nationallicence 
690 7 |a Lichenicidin  |2 nationallicence 
690 7 |a Synthetic biology  |2 nationallicence 
700 1 |a Kuthning  |D Anja  |u Institut für Chemie, Technische Universität Berlin, Straße des 17 Juni 124, 10623, Berlin, Germany  |4 aut 
700 1 |a Mösker  |D Eva  |u Institut für Chemie, Technische Universität Berlin, Straße des 17 Juni 124, 10623, Berlin, Germany  |4 aut 
700 1 |a Süssmuth  |D Roderich  |u Institut für Chemie, Technische Universität Berlin, Straße des 17 Juni 124, 10623, Berlin, Germany  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/15(2015-08-01), 6351-6361  |x 0175-7598  |q 99:15<6351  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-015-6557-6  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-015-6557-6  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Kuthning  |D Anja  |u Institut für Chemie, Technische Universität Berlin, Straße des 17 Juni 124, 10623, Berlin, Germany  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Mösker  |D Eva  |u Institut für Chemie, Technische Universität Berlin, Straße des 17 Juni 124, 10623, Berlin, Germany  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Süssmuth  |D Roderich  |u Institut für Chemie, Technische Universität Berlin, Straße des 17 Juni 124, 10623, Berlin, Germany  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/15(2015-08-01), 6351-6361  |x 0175-7598  |q 99:15<6351  |1 2015  |2 99  |o 253