Fusion with the cold-active esterase facilitates autotransporter-based surface display of the 10th human fibronectin domain in Escherichia coli.
Extremophiles
; 22(1): 141-150, 2018 Jan.
Article
em En
| MEDLINE
| ID: mdl-29256084
ABSTRACT
Cell surface display is a popular approach for the construction of whole-cell biocatalysts, live vaccines, and screening of combinatorial libraries. To develop a novel surface display system for the popular scaffold protein 10th human fibronectin type III domain (10Fn3) in Escherichia coli cells, we have used an α-helical linker and a C-terminal translocator domain from previously characterized autotransporter from Psychrobacter cryohalolentis K5T. The level of 10Fn3 passenger exposure at the cell surface provided by the hybrid autotransporter Fn877 and its C-terminal variants was low. To improve it, the fusion proteins containing 10Fn3 and the native autotransporter passenger Est877 or the cold-active esterase EstPc in different orientations were constructed and expressed as passenger domains. Using the whole-cell ELISA and activity assays, we have demonstrated that N-terminal position of EstPc in the passenger significantly improves the efficiency of the surface display of 10Fn3 in E. coli cells.
Palavras-chave
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Fibronectinas
/
Esterases
/
Sistemas de Secreção Tipo V
Limite:
Humans
Idioma:
En
Revista:
Extremophiles
Assunto da revista:
BIOLOGIA
Ano de publicação:
2018
Tipo de documento:
Article