Journal
BIOTECHNOLOGY LETTERS
Volume 36, Issue 12, Pages 2407-2416Publisher
SPRINGER
DOI: 10.1007/s10529-014-1642-y
Keywords
Biosynthesis; Directed evolution; Domain substitution; Module swapping; Non-ribosomal peptide synthetases; Rational engineering; Secondary metabolite
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Funding
- Royal Society of New Zealand Marsden Fund [VUW0901]
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Non-ribosomal peptide synthetases (NRPS) are large modular enzymes that govern the synthesis of numerous biotechnologically relevant products. Their mode of action is frequently compared to an assembly line, in which each module acts in a semi-autonomous but coordinated manner to add a specific monomer to a growing peptide chain, unfettered by ribosomal constraints. The modular nature of these systems offers tantalising prospects for synthetic biology, wherein the assembly line is re-engineered at a genetic level to generate a specific or combinatorial modified product. However, despite some success stories, a one size fits all approach to NRPS synthetic biology remains elusive. This review examines both rational and random mutagenesis strategies that have been employed to modify NRPS function, in an attempt to highlight key points that should be considered when seeking to re-engineer an NRPS biosynthetic template.
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