Directed evolution of the nonribosomal peptide synthetase AdmK generates new andrimid derivatives in vivo.
Chem Biol
; 18(5): 601-7, 2011 May 27.
Article
em En
| MEDLINE
| ID: mdl-21609841
ABSTRACT
Many lead compounds in the search for new drugs derive from peptides and polyketides whose similar biosynthetic enzymes have been difficult to engineer for production of new derivatives. Problems with generating multiple analogs in a single experiment along with lack of high-throughput methods for structure-based screening have slowed progress in this area. Here, we use directed evolution and a multiplexed assay to screen a library of >14,000 members to generate three derivatives of the antibacterial compound, andrimid. Another limiting factor in reengineering these mega-enzymes of secondary metabolism has been that commonly used hosts such as Escherichia coli often give lower product titers, so our reengineering was performed in the native producer, Pantoea agglomerans. This integrated in vivo approach can be extended to larger enzymes to create analogs of natural products for bioactivity testing.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Peptídeo Sintases
/
Evolução Molecular Direcionada
/
Antibacterianos
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article