Improvement of Heterologous Protein Production in Aspergillus oryzae by RNA Interference with .ALPHA.-Amylase Genes

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  • Improvement of heterologous protein production in Aspergillus oryzae by RNA interference with α-amylase genes
  • Improvement of heterologous protein production in Aspergillus oryzae by RNA interference with a amylase genes
  • Improvement of Heterologous Protein Production in<i>Aspergillus oryzae</i>by RNA Interference with α-Amylase Genes

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Abstract

Aspergillus oryzae RIB40 has three α-amylase genes (amyA, amyB, and amyC), and secretes α-amylase abundantly. However, large amounts of endogenous secretory proteins such as α-amylase can compete with heterologous protein in the secretory pathway and decrease its production yields. In this study, we examined the effects of suppression of α-amylase on heterologous protein production in A. oryzae, using the bovine chymosin (CHY) as a reporter heterologous protein. The three α-amylase genes in A. oryzae have nearly identical DNA sequences from those promoters to the coding regions. Hence we performed silencing of α-amylase genes by RNA interference (RNAi) in the A. oryzae CHY producing strain. The silenced strains exhibited a reduction in α-amylase activity and an increase in CHY production in the culture medium. This result suggests that suppression of α-amylase is effective in heterologous protein production in A. oryzae.

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