Recombinant Macrocyclic Lanthipeptides Incorporating Non-Canonical Amino Acids

Area of Science:

• Biochemistry
• Molecular Biology
• Synthetic Biology

Background:

• Nisin is a ribosomally synthesized lantibiotic with broad-spectrum antibacterial properties.
• Existing research aims to expand the structural diversity of lantibiotics like Nisin.
• Genetic incorporation of noncanonical amino acids (ncAAs) offers a route to novel Nisin variants.

Purpose of the Study:

• To achieve recombinant expression of Nisin variants incorporating ncAAs.
• To explore novel macrocyclic topologies in Nisin variants.
• To establish a methodology for creating diverse lanthipeptide variants with potentially enhanced activities.

Main Methods:

• Recombinant expression of the nisA gene, nisC, and nisB in Escherichia coli.
• Utilizing an orthogonal nonsense suppressor tRNA/aminoacyl-tRNA synthetase pair for ncAA incorporation.
• Genetic incorporation of ncAAs, including an α-chloroacetamide-containing ncAA, into the NisA structure.

Main Results:

• Successful recombinant expression of Nisin variants with genetically incorporated ncAAs.
• Demonstrated incorporation of an α-chloroacetamide-containing ncAA, leading to novel macrocyclic topologies.
• Established a versatile platform for generating structurally diverse Nisin variants.

Conclusions:

• The developed methodology enables the creation of Nisin variants with unique structural features.
• This approach facilitates the exploration of lanthipeptide variants with new or enhanced biological activities.
• The study provides a foundation for future research into Nisin and other lantibiotics.