ArticleMicrobial cell factories2020
Disruption or reduced expression of the orotidine-5'-decarboxylase gene pyrG increases citric acid production: a new discovery during recyclable genome editing in Aspergillus niger.
Article in Microbial cell factories, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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24 citing papers in PubMed, 38 citations in OpenAlex.
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- Constructing pyrG marker by CRISPR/Cas facilities the highly-efficient precise genome editing on industrial Aspergillus niger strain.Bioprocess and biosystems engineering · 2025Article
- Promoter engineering with programmable upstream activating sequences in Aspergillus Niger cell factory.Microbial cell factories · 2025Article
- Synthetic biology meets Aspergillus: engineering strategies for next-generation organic acid production.World journal of microbiology & biotechnology · 2025Review
- Reprogramming Metabolic Flux in Escherichia Coli to Enhance Chondroitin Production.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
- Utilization of CRISPR-Cas genome editing technology in filamentous fungi: function and advancement potentiality.Frontiers in microbiology · 2024Review
- Citric Acid: Properties, Microbial Production, and Applications in Industries.Molecules (Basel, Switzerland) · 2023Review
- Establishing a one-step marker-free CRISPR/Cas9 system for industrial Aspergillus niger using counter-selectable marker Ang-ace2.Biotechnology letters · 2023Article
- Transportation of citrinin is regulated by theJournal of Zhejiang University. Science. B · 2023Article
- Establishment, optimization, and application of genetic technology inFrontiers in microbiology · 2023Review
- Genome-wide transcription landscape of citric acid producingFrontiers in bioengineering and biotechnology · 2023Article
- Protocol for gene characterization in Aspergillus niger using 5S rRNA-CRISPR-Cas9-mediated Tet-on inducible promoter exchange.STAR protocols · 2022Article
- Comprehensively dissecting the hub regulation of PkaC on high-productivity and pellet macromorphology in citric acid producing Aspergillus niger.Microbial biotechnology · 2022Article
- Evaluation ofJournal of fungi (Basel, Switzerland) · 2022Article
- CRISPR/Cas9-Based Genome Editing and Its Application inJournal of fungi (Basel, Switzerland) · 2022Review
- Development of versatile and efficient genetic tools for the marine-derived fungus Aspergillus terreus RA2905.Current genetics · 2022Article
- Construction of gene modification system with highly efficient and markerless forFrontiers in microbiology · 2022Article
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
Abstract
backgroundAspergillus niger is a filamentous fungus used for the majority of global citric acid production. Recent developments in genome editing now enable biotechnologists to engineer and optimize A. niger. Currently, however, genetic-leads for maximizing citric acid titers in industrial A. niger isolates is limited.
resultsIn this study, we try to engineer two citric acid A. niger production isolates, WT-D and D353, to serve as platform strains for future high-throughput genome engineering. Consequently, we used genome editing to simultaneously disrupt genes encoding the orotidine-5'-decarboxylase (pyrG) and non-homologous end-joining component (kusA) to enable use of the pyrG selection/counter selection system, and to elevate homologous recombination rates, respectively. During routine screening of these pyrG mutant strains, we unexpectedly observed a 2.17-fold increase in citric acid production when compared to the progenitor controls, indicating that inhibition of uridine/pyrimidine synthesis may increase citric acid titers. In order to further test this hypothesis, the pyrG gene was placed under the control of a tetracycline titratable cassette, which confirmed that reduced expression of this gene elevated citric acid titers in both shake flask and bioreactor fermentation. Subsequently, we conducted intracellular metabolomics analysis, which demonstrated that pyrG disruption enhanced the glycolysis flux and significantly improved abundance of citrate and its precursors.
conclusionsIn this study, we deliver two citric acid producing isolates which are amenable to high throughput genetic manipulation due to pyrG/kusA deletion. Strikingly, we demonstrate for the first time that A. niger pyrG is a promising genetic lead for generating citric acid hyper-producing strains. Our data support the hypothesis that uridine/pyrimidine biosynthetic pathway offer future avenues for strain engineering efforts.
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