Metabolically independent and accurately adjustable Aspergillus sp. expression system.
Filamentous fungi are well-established expression hosts often used to produce extracellular proteins of use in the food and pharmaceutical industries. The expression systems presently used in Aspergillus species rely on either strong constitutive promoters, e.g., that for glyceraldehyde-3-phosphate dehydrogenase, or inducible systems derived from metabolic pathways, e.g., glaA (glucoamylase) or alc (alcohol dehydrogenase). We describe for Aspergillus nidulans and Aspergillus niger a novel expression system that utilizes the transcriptional activation of the human estrogen receptor by estrogenic substances. The system functions independently from metabolic signals and therefore can be used with low-cost, complex media. A combination of positive and negative regulatory elements in the promoter drives the expression of a reporter gene, yielding a linear dose response to the inducer. The off status is completely tight, yet the system responds within minutes to induction and reaches a level of expression of up to 15% of total cell protein after 8 h. Both Aspergillus species are very sensitive to estrogenic substances, and low-cost inducers function in the picomolar concentration range, at which estrogenic substances also can be found in the environment. Given this high sensitivity to estrogens, Aspergillus cells carrying estrogen-responsive units could be used to detect xenoestrogens in food or in the environment.
Pachlinger
Robert
R
Institut für Angewandte Genetik und Zellbiologie, BOKU-University of Natural Resources and Applied Life Sciences, Muthgasse 18, A-1190 Vienna, Austria.
Mitterbauer
Rudolf
R
Adam
Gerhard
G
Strauss
Joseph
J
D001233:Aspergillus nidulans / Q000235:genetics* / Q000378:metabolism; D001234:Aspergillus niger / Q000235:genetics* / Q000378:metabolism; D001709:Biotechnology / Q000379:methods; D003470:Culture Media; D004054:Diethylstilbestrol / Q000378:metabolism; D047628:Estrogen Receptor alpha / Q000235:genetics / Q000378:metabolism*; D004967:Estrogens / Q000032:analysis; D015966:Gene Expression Regulation, Fungal; D017930:Genes, Reporter; D005954:Glucosephosphate Dehydrogenase; D006801:Humans; D011401:Promoter Regions, Genetic; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D003470:Culture Media; D047628:Estrogen Receptor alpha; D004967:Estrogens; D004054:Diethylstilbestrol; D005954:Glucosephosphate Dehydrogenase; D001616:beta-Galactosidase