Isolation of a novel promoter for efficient protein production in Aspergillus oryzae.
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 beta-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.
Ishida
Hiroki
H
Research Institute, Gekkeikan Sake Co., Ltd, Fushimi-ku, Kyoto, Japan. h-ishida@gekkeikan.co.jp
Hata
Yoji
Y
Kawato
Akitsugu
A
Abe
Yasuhisa
Y
Kashiwagi
Yutaka
Y
D001236:Aspergillus oryzae / Q000235:genetics*; D016386:Chromosome Walking; D003001:Cloning, Molecular / Q000379:methods; D020224:Expressed Sequence Tags; D005285:Fermentation; D005966:Glucuronidase / Q000235:genetics; D037102:Lectins / Q000096:biosynthesis; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013482:Superoxide Dismutase / Q000235:genetics
D037102:Lectins; D011994:Recombinant Proteins; D013482:Superoxide Dismutase; D005966:Glucuronidase