Expanding the Genetic Code of Lactococcus lactis and Escherichia coli to Incorporate Non-canonical Amino Acids for Production of Modified Lantibiotics

Frontiers in Microbiology

Nediljko Budisa¹  Jessica H. Nickling¹  Tobias Baumann¹  Oscar P. Kuipers²  Maike Bartholomae²  David Peterhoff³  Ralf Wagner³ 

[1] Biocatalysis Group, Department of Chemistry, Technische Universität Berlin (Berlin Institute of Technology), Berlin, Germany;Department of Molecular Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, Netherlands;Institute of Medical Microbiology and Hygiene, Universität Regensburg, Regensburg, Germany;

关键词: stop codon suppression;    pyrrolysyl-tRNA synthetase;    nisin;    orthogonal translation system;    antimicrobial peptides;    bacteriocin;   

DOI  :  10.3389/fmicb.2018.00657

来源: DOAJ

【 摘 要 】

The incorporation of non-canonical amino acids (ncAAs) into ribosomally synthesized and post-translationally modified peptides, e.g., nisin from the Gram-positive bacterium Lactococcus lactis, bears great potential to expand the chemical space of various antimicrobials. The ncAA Nε-Boc-L-lysine (BocK) was chosen for incorporation into nisin using the archaeal pyrrolysyl-tRNA synthetase–tRNAPyl pair to establish orthogonal translation in L. lactis for read-through of in-frame amber stop codons. In parallel, recombinant nisin production and orthogonal translation were combined in Escherichia coli cells. Both organisms synthesized bioactive nisin(BocK) variants. Screening of a nisin amber codon library revealed suitable sites for ncAA incorporation and two variants displayed high antimicrobial activity. Orthogonal translation in E. coli and L. lactis presents a promising tool to create new-to-nature nisin derivatives.

【 授权许可】

Unknown