High-level synthesis of recombinant HIV-1 protease and the recovery of active enzyme from inclusion bodies

Gene. 1990 Mar 15;87(2):243-8. doi: 10.1016/0378-1119(90)90308-e.


A complete chemical synthesis and assembly of genes for the production of human immunodeficiency virus type-I protease (HIV-PR) and its precursors are described. The T7 expression system was used to produce high levels of active HIV-PR and its precursors in Escherichia coli inclusion bodies. The gene encoding the open reading frames of HIV-PR was expressed in E. coli as a 10-kDa protein, while the genes encoding HIV-PR precursors were expressed as larger proteins, which were partially processed in E. coli to the 10-kDa form. These processing events are autoproteolytic, since a single-base mutation, changing the active-site aspartic acid to glycine, completely abolished the conversion. HIV-PR can be released with 8 M urea from washed cellular inclusion bodies, resulting in a preparation with few bacterial host proteins. After refolding, this preparation contains no nonspecific protease or peptidase activities. The recombinant HIV-PR isolated from inclusion bodies cleaves HIV-PR substrates specifically with a specific activity comparable to column-purified HIV-PR.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Endopeptidases / biosynthesis
  • Endopeptidases / genetics*
  • Endopeptidases / isolation & purification
  • Endopeptidases / metabolism
  • Enzyme Precursors / biosynthesis
  • Enzyme Precursors / genetics
  • Escherichia coli / genetics*
  • Gene Expression*
  • Gene Products, pol / biosynthesis
  • Gene Products, pol / genetics*
  • Gene Products, pol / isolation & purification
  • Gene Products, pol / metabolism
  • Genes, Synthetic*
  • Genes, Viral
  • HIV Protease
  • HIV-1 / enzymology*
  • Inclusion Bodies / enzymology
  • Molecular Sequence Data
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Sequence Homology, Nucleic Acid


  • Enzyme Precursors
  • Gene Products, pol
  • Recombinant Proteins
  • Endopeptidases
  • HIV Protease

Associated data

  • GENBANK/M36450