Efficient recombinant expression of SFTI-1 in bacterial cells using intein-mediated protein trans-splicing.
Biopolymers
; 106(6): 818-824, 2016 Nov.
Article
em En
| MEDLINE
| ID: mdl-27178003
ABSTRACT
We report for the first time the recombinant expression of bioactive wild-type sunflower trypsin inhibitor 1 (SFTI-1) inside E. coli cells by making use of intracellular protein trans-splicing in combination with a high efficient split-intein. SFTI-1 is a small backbone-cyclized polypeptide with a single disulfide bridge and potent trypsin inhibitory activity. Recombinantly produced SFTI-1 was fully characterized by NMR and was observed to actively inhibit trypsin. The in-cell expression of SFTI-1 was very efficient reaching intracellular concentration ≈ 40 µM. This study clearly demonstrates the possibility of generating genetically encoded SFTI-based peptide libraries in live E. coli cells, and is a critical first step for developing in-cell screening and directed evolution technologies using the cyclic peptide SFTI-1 as a molecular scaffold. © 2016 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 106 818-824, 2016.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Peptídeos Cíclicos
/
Expressão Gênica
/
Processamento de Proteína
/
Inteínas
/
Helianthus
Idioma:
En
Ano de publicação:
2016
Tipo de documento:
Article