       Document 0464
 DOCN  M9640464
 TI    Large-scale production of HIV-1 protease from Escherichia coli using
       selective extraction and membrane fractionation.
 DT    9604
 AU    Gustafson ME; Junger KD; Foy BA; Baez JA; Bishop BF; Rangwala SH;
       Michener ML; Leimgruber RM; Houseman KA; Mueller RA; et al; G.D. Searle
       and Co., Chesterfield, Missouri 63198, USA.
 SO    Protein Expr Purif. 1995 Aug;6(4):512-8. Unique Identifier : AIDSLINE
       MED/96136639
 AB    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 40:60 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.
 DE    Acetic Acids  Amino Acid Sequence  Escherichia coli/*GENETICS
       Fermentation  Fractionation  Genetic Vectors  HIV
       Protease/*GENETICS/*ISOLATION & PURIF  Inclusion Bodies/ENZYMOLOGY
       Intracellular Membranes/ENZYMOLOGY  Molecular Sequence Data  Plasmids
       Recombinant Fusion Proteins/GENETICS/ISOLATION & PURIF  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

