Large-scale production of HIV-1 protease from Escherichia coli using selective extraction and membrane fractionation.
Protein Expr Purif
; 6(4): 512-8, 1995 Aug.
Article
em En
| MEDLINE
| ID: mdl-8527938
ABSTRACT
Human immunodeficiency virus type 1 (HIV-1) protease was expressed in Escherichia coli as a fusion protein with the N-terminal sequence of IGF-2. The protein accumulated in inclusion bodies as a 4060 mixture of unprocessed fusion protein and processed protein. A simple purification procedure was developed that yielded 30-40 mg of active protease per liter of fermentation broth with a recovery of 30-40%. The purification process involved the selective extraction of HIV-1 protease from E. coli inclusion bodies with 50% acetic acid and fractional diafiltration to remove impurities and low-molecular-weight protease-related fragments. No chromatographic steps were employed, yet the HIV-1 protease produced by this procedure was greater than 95% pure by SDS-PAGE, reverse-phase HPLC, and N-terminal sequence analysis.
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Coleções:
01-internacional
Contexto em Saúde:
3_ND
Base de dados:
MEDLINE
Assunto principal:
Protease de HIV
/
Escherichia coli
Idioma:
En
Revista:
Protein Expr Purif
Ano de publicação:
1995
Tipo de documento:
Article