Isolation of a novel promoter for efficient protein expression by <Emphasis Type="Italic">Aspergillus oryzae</Emphasis> in solid-state culture |
| |
Authors: | Hiroki Bando Hiromoto Hisada Hiroki Ishida Yoji Hata Yoshio Katakura Akihiko Kondo |
| |
Institution: | (1) Research Institute, Gekkeikan Sake Co. Ltd., 101 Shimotoba-koyanagi-cho, Fushimi-ku, Kyoto 612-8385, Japan;(2) Department of Life Science and Bioengineering, Faculty of Chemistry, Materials and Bioengineering, Kansai University, 3-3-35 Tamate-cho, Suita-shi, Osaka 564-8680, Japan;(3) Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan |
| |
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. |
| |
Keywords: | |
本文献已被 PubMed SpringerLink 等数据库收录! |
|