A Subdomain Swap Strategy for Reengineering Nonribosomal Peptides
| Autor: | Hajo Kries, David L. Niquille, Donald Hilvert |
|---|---|
| Jazyk: | angličtina |
| Předmět: |
Clinical Biochemistry
Peptide Gramicidin S Biology Protein Engineering Biochemistry Mass Spectrometry chemistry.chemical_compound Protein structure Nonribosomal peptide Drug Discovery Amino Acid Sequence Peptide Synthases Molecular Biology Peptide sequence Adenylylation Amino Acid Isomerases Pharmacology chemistry.chemical_classification General Medicine Protein engineering Recombinant Proteins Protein Structure Tertiary chemistry Molecular Medicine Peptides Swap (computer programming) |
| Zdroj: | Chemistry & Biology. (5):640-648 |
| ISSN: | 1074-5521 |
| DOI: | 10.1016/j.chembiol.2015.04.015 |
| Popis: | SummaryNonribosomal peptide synthetases (NRPSs) protect microorganisms from environmental threats by producing diverse siderophores, antibiotics, and other peptide natural products. Their modular molecular structure is also attractive from the standpoint of biosynthetic engineering. Here we evaluate a methodology for swapping module specificities of these mega-enzymes that takes advantage of flavodoxin-like subdomains involved in substrate recognition. Nine subdomains encoding diverse specificities were transplanted into the Phe-specific GrsA initiation module of gramicidin S synthetase. All chimeras could be purified as soluble protein. One construct based on a Val-specific subdomain showed sizable adenylation activity and functioned as a Val-Pro diketopiperazine synthetase upon addition of the proline-specific GrsB1 module. These results suggest that subdomain swapping could be a viable alternative to previous NRPS design approaches targeting binding pockets, domains, or entire modules. The short length of the swapped sequence stretch may facilitate straightforward exploitation of the wealth of existing NRPS modules for combinatorial biosynthesis. |
| Databáze: | OpenAIRE |
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