Mapping the Nonribosomal Specificity Code through Promiscuity-Guided A-Domain Engineering

Key Points

• The aim is to understand how nonribosomal peptide synthetases can be engineered to change their specificity towards different building blocks.
• Analysis of binding pocket residues responsible for specificity in nonribosomal peptide synthetases.
• Engineering of A-domains to assess changes in building block preferences.
• Comparison of engineered constructs to evaluate specificity outcomes.
• Identified key residues that influence binding pocket specificity.
• Demonstrated that certain engineered A-domains show altered specificity for various building blocks.
• Provided insights into the design of novel NRPS products.