Production of recombinant Der fI (a major mite allergen) by Aspergillus oryzae

Biosci Biotechnol Biochem. 1999 Apr;63(4):703-9. doi: 10.1271/bbb.63.703.

Abstract

Der fI is a major mite allergen. To produce Der fI by Aspergillus oryzae, we placed a DNA fragment encoding precursor-type recombinant Der fI E(-1)K (reDer fI E(-1) K), which had the C-terminal amino acid of the pro-sequence (Glu) changed to Lys, downstream of the glaA gene promoter and introduced it into Aspergillus oryzae. In liquid culture, most of the reDer fI E(-1)K produced by the transformants was degraded when culture was shaken vigorously. However, the degradation of reDer fI E(-1)K was suppressed when it was shaken gently. The processed reDer fI E(-1)K could be obtained after lysylendopeptidase and endoglycosidase Hf (Endo Hf) treatment. The yield of processed reDer fI E(-1)K was 8 mg/l. When the transformant was grown on a wheat bran culture, the yield of processed reDer fI E(-1)K reached 48 mg/kg. Because processed reDer fI E(-1)Ks obtained from both cultures had almost the same IgE-binding activity and elicited the same skin reaction as native Der fI, they could be very useful for diagnostic purposes or immunotherapy.

Publication types

  • Clinical Trial

MeSH terms

  • Allergens / biosynthesis*
  • Animals
  • Antigens, Dermatophagoides
  • Aspergillus oryzae / genetics
  • Aspergillus oryzae / metabolism*
  • Blotting, Southern
  • Culture Media
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Glycoproteins / biosynthesis*
  • Glycoproteins / chemistry
  • Humans
  • Mites / immunology*
  • Plasmids / genetics
  • Recombinant Proteins / biosynthesis
  • Skin Tests
  • Transformation, Bacterial
  • Trichloroacetic Acid

Substances

  • Allergens
  • Antigens, Dermatophagoides
  • Culture Media
  • Glycoproteins
  • Recombinant Proteins
  • Trichloroacetic Acid