2004 Volume 68 Issue 9 Pages 1849-1857
A novel protein overexpression system of Aspergillus oryzae was constructed. Five promoters which originate from A. oryzae expressed sequence tag (EST) clones in submerged culture were obtained by genome walking. These were subjected to β-glucuronidase (GUS) reporter assays. The promoter of manganese superoxide dismutase-encoding gene (sodM) showed the most GUS production. The sodM gene was abundantly expressed in submerged culture but little expressed in solid-state culture. The sodM promoter was approximately 3-fold induced by the addition of 0.01% H2O2. Glucoamylase production in A. oryzae using the sodM promoter led to secretion of approximately 1 g/l-broth in Czapek-Dox medium for 3 d. Fucose lectin production in A. oryzae using the sodM promoter led to overexpression as a specific and abundant intracellular protein.
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