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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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- 3 Web of Science
- 4 Crossref
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Abstract
- 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.
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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
- Construction of an Aspergillus oryzae △nptB△pyrG Host for Homologous Expression of Lipase and Catalytic Property Characterization of Recombinant Lipase
Research Support, Non-U.S. Gov't
- The Pectate Lyase Encoded by the pecCl1 Gene Is an Important Determinant for the Aggressiveness of Colletotrichum lindemuthianum
- Andréia Cnossen-Fassoni , Denise Mara Soares Bazzolli , Sérgio Hermínio Brommonschenkel , Elza Fernandes de Araújo , Marisa Vieira de Queiroz
- J. Microbiol. 2013;51(4):461-470. Published online August 30, 2013
- DOI: https://doi.org/10.1007/s12275-013-3078-9
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- 14 Scopus
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Abstract
- Colletotrichum lindemuthianum is the causal agent of anthracnose in the common bean, and the genes that encode its cell-wall-degrading enzymes are crucial for the development of the disease. Pectinases are the most important group of cell wall-degrading enzymes produced by phytopathogenic fungi. The pecC1l gene, which encodes a pectate lyase in C. lindemuthianum, was isolated and characterized. Possible cis-regulatory elements and transcription factor binding sites that may be involved in the regulation of genetic expression were detected in the promoter region of the gene. pecCl1 is represented by a single copy in the genome of C. lindemuthianum, though in silico analyses of the genomes of Colletotrichum graminicola and Colletotrichum higginsianum suggest that the genome of C. lindemuthianum includes other genes that encode pectate lyases. Phylogenetic analysis detected two groups that clustered based on different members of the pectate lyase family. Analysis of the differential expression of pecCl1 during different stages of infection showed a significant increase in pecCl1 expression five days after infection, at the onset of the necrotrophic phase. The split-maker technique proved to be an efficient method for inactivation of the pecCl1 gene, which allowed functional study of a mutant with a site-specific integration. Though gene inactivation did not result in complete loss of pectate lyase activity, the symptoms of anthracnose were reduced. Analysis of pectate lyases might not only contribute to the understanding of anthracnose in the common bean but might also lead to the discovery of an additional target for controlling anthracnose.
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