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

• Published in: Applied microbiology and biotechnology Vol. 99; no. 15; pp. 6351 - 6361
• Main Authors: Kuthning, Anja, Mösker, Eva, Süssmuth, Roderich D
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.08.2015; Springer; Springer Nature B.V
• Subjects: Amino acids; Analysis; Antibiotics; Antimicrobial agents; Applied Genetics and Molecular Biotechnology; Bacillus licheniformis; bacteriocins; Bacteriocins - genetics; Bacteriocins - metabolism; Bioavailability; Biomedical and Life Sciences; Biotechnology; Blias; Cloning; Cloning, Molecular; E coli; engineering; Enzymes; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Gene Expression; Genes; Genetic engineering; Gram-positive bacteria; heterologous gene expression; Life Sciences; Metabolic Engineering - methods; Metabolic Networks and Pathways - genetics; Microbial Genetics and Genomics; Microbiology; Mutagenesis; Natural products; peptide antibiotics; Peptides; Pharmaceutical industry; Pharmaceutical sciences; Plasmids; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Studies; translation (genetics); Yeast; yields; Genetic engineering; Lantibiotic; Synthetic biology; Lichenicidin; Heterologous expression
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-015-6557-6

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 4 mg L⁻¹ for Bliα and 6 mg L⁻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.