Production of recombinant Der fI (a major mite allergen) by Aspergillus oryzae.
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.
Shoji
H
H
Institute for Production Research and Development, Nikka Whisky Distilling Co., Ltd., Chiba, Japan. XLR03635@niftyserve.or.jp
Horiuchi
H
H
Takagi
M
M
D000485:Allergens / Q000096:biosynthesis*; D000818:Animals; D039741:Antigens, Dermatophagoides; D001236:Aspergillus oryzae / Q000235:genetics / Q000378:metabolism*; D015139:Blotting, Southern; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006023:Glycoproteins / Q000096:biosynthesis* / Q000737:chemistry; D006801:Humans; D008925:Mites / Q000276:immunology*; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D012882:Skin Tests; D014169:Transformation, Bacterial; D014238:Trichloroacetic Acid
D000485:Allergens; D039741:Antigens, Dermatophagoides; D003470:Culture Media; D006023:Glycoproteins; D011994:Recombinant Proteins; D014238:Trichloroacetic Acid