Transcriptional regulation has been adopted for developing metabolic engineering tools. The regulatory promoter is a crucial
genetic element for strain optimization. In this study, a gene set of Aspergillus oryzae with highly constitutive expression
across different growth stages was identified through transcriptome data analysis. The candidate promoters were functionally
characterized in A. oryzae by transcriptional control of β-glucuronidase (GUS) as a reporter. The results showed that
the glyceraldehyde triphosphate dehydrogenase promoter (PgpdA1) of A. oryzae with a unique structure displayed the most
robust strength in constitutively controlling the expression compared to the PgpdA2 and other putative promoters tested. In
addition, the ubiquitin promoter (Pubi) of A. oryzae exhibited a moderate expression strength. The deletion analysis revealed
that the 5' untranslated regions of gpdA1 and ubi with the length of 1028 and 811 nucleotides, counted from the putative
translation start site (ATG), respectively, could efficiently drive the GUS expression. Interestingly, both promoters could
function on various carbon sources for cell growth. Glucose was the best fermentable carbon source for allocating high constitutive
expressions during cell growth, and the high concentrations (6–8% glucose, w/v) did not repress their functions. It
was also demonstrated that the secondary metabolite gene coding for indigoidine could express under the control of PgpdA1
or Pubi promoter. These strong and moderate promoters of A. oryzae provided beneficial options in tuning the transcriptional
expression for leveraging the metabolic control towards the targeted products.
Citations
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