Applied Microbiology and Biotechnology, Vol.92, No.3, 561-569, 2011
Isolation of a novel promoter for efficient protein expression by Aspergillus oryzae in solid-state culture
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-beta-glucanase from A. oryzae (CelA) as the reporter, promoter activity was found to be higher than that of the alpha-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 30A degrees C or 37A degrees 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:Hemolysin-like protein-encoding gene (hlyA);Aspergillus oryzae;Solid-state culture;Endo-1,4-beta-glucanase;Promoter