title: Isolation of a novel promoter for efficient protein expression by Aspergillus oryzae in solid-state culture.
abstract: A novel promoter from a hemolysin-like protein encoding the gene, hlyA, was characterized for protein overexpression in Aspergillus oryzae grown in solid-state culture. Using endo-1,4-β-glucanase from A. oryzae (CelA) as the reporter, promoter activity was found to be higher than that of the α-amylase (amyA) and manganese superoxide dismutase (sodM) genes not only in wheat bran solid-state culture but also in liquid culture. Expression of the A. oryzae endoglucanase CelB and two heterologous endoglucanases (TrEglI and TrEglIII from Trichoderma reesei) under the control of the hlyA promoter were also found to be stronger than under the control of the amyA promoter in A. oryzae grown in wheat bran solid-state culture, suggesting that the hlyA promoter may be useful for the overproduction of other proteins as well. In wheat bran solid-state culture, the productivity of the hlyA promoter in terms of protein produced was high when the cultivation temperature was 30°C or 37°C, when the water content was 0.6 or 0.8 ml/g wheat bran, and from 48 to 72 h after inoculation. Because A. oryzae sporulated actively under these conditions and because hemolysin has been reported to play a role in fungal fruiting body formation, high-level expression of hlyA may be related to sporulation.
authors: Research Institute, Gekkeikan Sake Co. Ltd., 101 Shimotoba-koyanagi-cho, Fushimi-ku, Kyoto, 612-8385, Japan. bandou@gekkeikan.co.jp
authors: Hiroki
authors: H
authors: Bando
authors: Hiromoto
authors: H
authors: Hisada
authors: Hiroki
authors: H
authors: Ishida
authors: Yoji
authors: Y
authors: Hata
authors: Yoshio
authors: Y
authors: Katakura
authors: Akihiko
authors: A
authors: Kondo
mesh_terms: D001236:Aspergillus oryzae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D001483:Base Sequence; D002480:Cellulase / Q000096:biosynthesis* / Q000235:genetics; D004043:Dietary Fiber / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013696:Temperature; D013997:Time Factors; D014242:Trichoderma / Q000201:enzymology / Q000235:genetics
chemical_list: D004043:Dietary Fiber; D011994:Recombinant Proteins; D002480:Cellulase