Enabling low cost biopharmaceuticals: high level interferon alpha-2b production in Trichoderma reesei.
Microb Cell Fact
; 15(1): 104, 2016 Jun 10.
Article
em En
| MEDLINE
| ID: mdl-27287473
ABSTRACT
BACKGROUND:
The filamentous fungus Trichoderma reesei has tremendous capability to secrete over 100 g/L of proteins and therefore it would make an excellent host system for production of high levels of therapeutic proteins at low cost. We have developed T. reesei strains suitable for production of therapeutic proteins by reducing the secreted protease activity. Protease activity has been the major hindrance to achieving high production levels. We have constructed a series of interferon alpha-2b (IFNα-2b) production strains with 9 protease deletions to gain knowledge for further strain development.RESULTS:
We have identified two protease deletions that dramatically improved the production levels. Deletion of the subtilisin protease slp7 and the metalloprotease amp2 has enabled production levels of IFNα-2b up to 2.1 and 2.4 g/L, respectively. With addition of soybean trypsin protease inhibitor the level of production improved to 4.5 g/L, with an additional 1.8 g/L still bound to the secretion carrier protein.CONCLUSIONS:
High levels of IFNα-2b were produced using T. reesei strains with reduced protease secretion. Further strain development can be done to improve the production system by reducing protease activity and improving carrier protein cleavage.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Trichoderma
/
Proteínas Recombinantes
/
Interferon-alfa
Tipo de estudo:
Health_economic_evaluation
Idioma:
En
Revista:
Microb Cell Fact
Ano de publicação:
2016
Tipo de documento:
Article