Nonribosomal biosynthesis of backbone-modified peptides
| Autor: | David L. Niquille, Takahiro Mori, David Fercher, Douglas A. Hansen, Hajo Kries, Donald Hilvert |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Peptide Biosynthesis General Chemical Engineering Protein Engineering 01 natural sciences Ribosome 03 medical and health sciences chemistry.chemical_compound Fluorescence-Activated Cell Sorting Biosynthesis Nonribosomal peptide Peptide Synthases chemistry.chemical_classification DNA ligase Molecular Structure 010405 organic chemistry General Chemistry Protein engineering Yeast 0104 chemical sciences 030104 developmental biology chemistry Biochemistry Biocatalysis Peptides Ribosomes |
| Zdroj: | Nature chemistry. 10(3) |
| ISSN: | 1755-4349 |
| Popis: | Biosynthetic modification of nonribosomal peptide backbones represents a potentially powerful strategy to modulate the structure and properties of an important class of therapeutics. Using a high-throughput assay for catalytic activity, we show here that an L-Phe-specific module of an archetypal nonribosomal peptide synthetase can be reprogrammed to accept and process the backbone-modified amino acid (S)-β-Phe with near-native specificity and efficiency. A co-crystal structure with a non-hydrolysable aminoacyl-AMP analogue reveals the origins of the 40,000-fold α/β-specificity switch, illuminating subtle but precise remodelling of the active site. When the engineered catalyst was paired with downstream module(s), (S)-β-Phe-containing peptides were produced at preparative scale in vitro (~1 mmol) and high titres in vivo (~100 mg l |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink