- Transcriptome‑based Mining of the Constitutive Promoters for Tuning Gene Expression in Aspergillus oryzae
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Kobkul Laoteng , Jutamas Anantayanon , Chanikul Chutrakul , Sarocha Panchanawaporn , Sukanya Jeennor
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J. Microbiol. 2023;61(2):199-210. Published online February 6, 2023
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DOI: https://doi.org/10.1007/s12275-023-00020-0
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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  - Regulatory mechanisms underlying stress-induced accumulation of plant secondary metabolites
Sujing He, Jiangna Wang, Ning Yang, Hongwei Li, Kunqian Li, Lin Li, Wei’e Wen, Tingting Luo, Juanjuan Zhao, Delin Xu
Journal of Applied Research on Medicinal and Aromatic Plants.2026; 50: 100689. CrossRef - Two-Step Engineering of Food-Grade Aspergillus oryzae via Endogenous Signal Peptides and Vesicle Trafficking Proteins to Enhance Carrier-Free Protein Secretion
Sarocha Panchanawaporn, Nakul Rattanaphan, Sukanya Jeennor, Jutamas Anantayanon, Weerapong Woraprayote, Laphaslada Pumpuang, Thipphiya Karirat, Nuttamon Prompakdee, Kobkul Laoteng, Chanikul Chutrakul
Journal of Fungi.2026; 12(4): 289. CrossRef - Mining and characterization of endogenous constitutive promoters in Aspergillus oryzae
Sha Huang, Laimao Luo, Jingyin Zhou, Zhui Tu, Qinhua He, Yanping Li
Food Bioscience.2026; 81: 109086. CrossRef - Bacillus subtilis expressing double‐stranded RNA induced RNA interference in Tribolium castaneum
Li He, Yifan Zhou, Junhua Xie, Yucheng Gu, Yanna Huang, Xueming Tang
Pest Management Science.2026; 82(10): 10073. CrossRef - 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.2025; 197(2): 873. CrossRef - Transcriptome-Based Mining of the Strong Promoters for Hyperproduction of Gibberellin GA3 in Fusarium fujikuroi
Qi Guo, Yue-Feng Zhong, Xin-Yu Chen, Ya-Wen Li, Yu-Xin Yang, Zhi-Kui Nie, Tian-Qiong Shi
Journal of Agricultural and Food Chemistry.2025; 73(14): 8440. CrossRef - Promoter engineering of filamentous fungi for novel natural product discovery
Xiangzhou Gong, Jing Tian, Huawei Zhang
Bioorganic Chemistry.2025; 163: 108798. 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 - Development of Aspergillus oryzae BCC7051 as a Robust Cell Factory Towards the Transcriptional Regulation of Protease-Encoding Genes for Industrial Applications
Sarocha Panchanawaporn, Chanikul Chutrakul, Sukanya Jeennor, Jutamas Anantayanon, Kobkul Laoteng
Journal of Fungi.2024; 11(1): 6. 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
- Promoter exchange of the cryptic nonribosomal peptide synthetase gene for oligopeptide production in Aspergillus oryzae
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Chanikul Chutrakul , Sarocha Panchanawaporn , Sukanya Jeennor , Jutamas Anantayanon , Kobkul Laoteng
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J. Microbiol. 2022;60(1):47-56. Published online November 9, 2021
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DOI: https://doi.org/10.1007/s12275-022-1442-3
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Abstract
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Oligopeptides with functional activities are of current interest
in the nutraceutical and medical sectors. The development of
the biosynthetic process of oligopeptides through a nonribosomal
peptide synthetase (NRPS) system has become more
challenging. To develop a production platform for nonribosomal
peptides (NRPs), reprogramming of transcriptional
regulation of the acv gene encoded ACV synthetase (ACVS)
was implemented in Aspergillus oryzae using the CRISPRCas9
system. Awakening silent acv expression was successfully
achieved by promoter substitution. Among the three exchanged
promoters, AoPgpdA, AoPtef1, and PtPtoxA, the
replacement of the native promoter with AoPgpdA led to the
highest ACV production in A. oryzae. However, the ACV production
of the AoPGpdA strain was also dependent on the
medium composition, in which urea was the best nitrogen
source, and a C:N ratio of 20:1 was optimal for tripeptide production.
In addition to cell growth, magnesium ions are an
essential element for ACV production and might participate
in ACVS activity. It was also found that ACV was the growthassociated
product of the engineered strain that might be a
result
of constitutive transcriptional control by the AoPgpdA
promoter. This study offers a potential strategy for nonribosomal
ACV production using the fungal system, which is applicable
for redesigning bioactive oligopeptides with industrial
relevance.
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Citations
Citations to this article as recorded by - Two-Step Engineering of Food-Grade Aspergillus oryzae via Endogenous Signal Peptides and Vesicle Trafficking Proteins to Enhance Carrier-Free Protein Secretion
Sarocha Panchanawaporn, Nakul Rattanaphan, Sukanya Jeennor, Jutamas Anantayanon, Weerapong Woraprayote, Laphaslada Pumpuang, Thipphiya Karirat, Nuttamon Prompakdee, Kobkul Laoteng, Chanikul Chutrakul
Journal of Fungi.2026; 12(4): 289. CrossRef - Advances in Fungal Promoter Engineering for Enhancing Secondary Metabolite Biosynthesis
Nan Pu, Huawei Zhang
Biotechnology Journal.2025;[Epub] CrossRef - CRISPR-Cas9 Gene Editing in Aspergillus: From Pathogenesis to Metabolic Engineering
Danni Hu, Ruoyu Zhao, Yingxu Lin, Chunmiao Jiang
Biology.2025; 15(1): 53. CrossRef - Strategies for the Enhancement of Secondary Metabolite Production via Biosynthesis Gene Cluster Regulation in Aspergillus oryzae
Xiao Jia, Jiayi Song, Yijian Wu, Sai Feng, Zeao Sun, Yan Hu, Mengxue Yu, Rui Han, Bin Zeng
Journal of Fungi.2024; 10(5): 312. CrossRef - Development of Aspergillus oryzae BCC7051 as a Robust Cell Factory Towards the Transcriptional Regulation of Protease-Encoding Genes for Industrial Applications
Sarocha Panchanawaporn, Chanikul Chutrakul, Sukanya Jeennor, Jutamas Anantayanon, Kobkul Laoteng
Journal of Fungi.2024; 11(1): 6. CrossRef - Transcriptome-based Mining of the Constitutive Promoters for Tuning Gene Expression in Aspergillus oryzae
Kobkul Laoteng, Jutamas Anantayanon, Chanikul Chutrakul, Sarocha Panchanawaporn, Sukanya Jeennor
Journal of Microbiology.2023; 61(2): 199. 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 - Synthetic microbes and biocatalyst designs in Thailand
Duangthip Trisrivirat, Ruchanok Tinikul, Pimchai Chaiyen
Biotechnology Notes.2023; 4: 28. CrossRef - Potential of Aspergillus oryzae as a biosynthetic platform for indigoidine, a non-ribosomal peptide pigment with antioxidant activity
Sarocha Panchanawaporn, Chanikul Chutrakul, Sukanya Jeennor, Jutamas Anantayanon, Nakul Rattanaphan, Kobkul Laoteng, Daniel Cullen
PLOS ONE.2022; 17(6): e0270359. CrossRef - CRISPR/Cas9-Based Genome Editing and Its Application in Aspergillus Species
Feng-Jie Jin, Bao-Teng Wang, Zhen-Dong Wang, Long Jin, Pei Han
Journal of Fungi.2022; 8(5): 467. CrossRef
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