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Transcriptome‑based Mining of the Constitutive Promoters for Tuning Gene Expression in Aspergillus oryzae
Kobkul Laoteng , Jutamas Anantayanon , Chanikul Chutrakul , Sarocha Panchanawaporn , Sukanya Jeennor
J. Microbiol. 2023;61(2):199-210.   Published online February 6, 2023
DOI: https://doi.org/10.1007/s12275-023-00020-0
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AbstractAbstract
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

Citations to this article as recorded by  
  • Construction of an Aspergillus oryzae △nptB△pyrG Host for Homologous Expression of Lipase and Catalytic Property Characterization of Recombinant Lipase
    Yueting Zhang, Hongmei Nie, Fei Zhang, Mengmeng Jin, Zhao Wang, Jianyong Zheng
    Applied Biochemistry and Biotechnology.2024;[Epub]     CrossRef
  • Mining and Understanding of New Transcriptional Regulatory Elements from Licorice-Derived Endophyte Serratia Rubidaea W12-1
    Ying Zhang, Yunyang Ma, Bing Hu, H.M. Zabed, A.K. Singh, M.A. Ibrahim, N. Chen
    BIO Web of Conferences.2024; 142: 03018.     CrossRef
  • Exploring and Engineering Novel Strong Promoters for High-Level Protein Expression in Bacillus subtilis DB104 through Transcriptome Analysis
    Ji-Su Jun, Hyang-Eun Jeong, Kwang-Won Hong
    Microorganisms.2023; 11(12): 2929.     CrossRef
  • Efficient de novo production of bioactive cordycepin by Aspergillus oryzae using a food-grade expression platform
    Sukanya Jeennor, Jutamas Anantayanon, Sarocha Panchanawaporn, Chanikul Chutrakul, Wanwipa Vongsangnak, Kobkul Laoteng
    Microbial Cell Factories.2023;[Epub]     CrossRef

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