Global Transcriptional Response of Aspergillus niger to Blocked Active Citrate Export through Deletion of the Exporter Gene

Laothanachareon, Thanaporn; Bruinsma, Lyon; Nijsse, Bart; Schonewille, Tom; Suárez-Diez, María; Tamayo-Ramos, Juan Antonio; Santos, Vítor A. P. Martins dos; Schaap, Peter J.

doi:10.3390/jof7060409

Cited by 7 publications

(5 citation statements)

References 52 publications

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“…We observed that CexA overexpression dramatically influenced the transcriptional pattern of genes involved in carbohydrate transport and metabolism ( Figure S4 ), suggesting its impacts on carbon metabolic flux redistribution. Three glucose transporters MstC, MstG, and MstH displayed the similar expression pattern as the cexA gene, which was consistent with the previous transcriptional analysis [ 47 ]. Additionally, we also discovered that several genes beneficial for intracellular citric acid accumulation were significantly regulated, including starch hydrolysis, precursor supply, mitochondrial citrate/malate shuttle, alternative oxidase respiratory chain, citric acid degradation, and by-product biosynthesis.…”

Section: Discussionsupporting

confidence: 90%

Evaluation of Aspergillus niger Six Constitutive Strong Promoters by Fluorescent-Auxotrophic Selection Coupled with Flow Cytometry: A Case for Citric Acid Production

Zheng

Wang

et al. 2022

JoF

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Aspergillus niger is an important industrial workhorse for the biomanufacturing of organic acids, proteins, etc. Well-controlled genetic regulatory elements, including promoters, are vital for strain engineering, but available strong promoters for A. niger are limited. Herein, to efficiently assess promoters, we developed an accurate and intuitive fluorescent-auxotrophic selection workflow based on mCherry, pyrG, CRISPR/Cas9 system, and flow cytometry. With this workflow, we characterized six endogenous constitutive promoters in A. niger. The endogenous glyceraldehyde-3-phosphate dehydrogenase promoter PgpdAg showed a 2.28-fold increase in promoter activity compared with the most frequently used strong promoter PgpdAd from A. nidulans. Six predicted conserved motifs, including the gpdA-box, were verified to be essential for the PgpdAg activity. To demonstrate its application, the promoter PgpdAg was used for enhancing the expression of citrate exporter cexA in a citric acid-producing isolate D353.8. Compared with the cexA controlled by PgpdAd, the transcription level of the cexA gene driven by PgpdAg increased by 2.19-fold, which is consistent with the promoter activity assessment. Moreover, following cexA overexpression, several genes involved in carbohydrate transport and metabolism were synergically upregulated, resulting in up to a 2.48-fold increase in citric acid titer compared with that of the parent strain. This study provides an intuitive workflow to speed up the quantitative evaluation of A. niger promoters and strong constitutive promoters for fungal cell factory construction and strain engineering.

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Section: Discussionsupporting

confidence: 90%

Evaluation of Aspergillus niger Six Constitutive Strong Promoters by Fluorescent-Auxotrophic Selection Coupled with Flow Cytometry: A Case for Citric Acid Production

Zheng

Wang

et al. 2022

JoF

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“…Shi et al 102 stated that increasing fungal biomass during fermentation resulted in increasing the protein content of the end products. The ability of microorganisms to secrete a variety of enzymes, 103 biotransformation of soluble carbohydrate into microbial protein, 104 secretion of some extracellular components, 105 and free hydrolyzed peptides that are proteinaceous in nature, 106 might result in an increase in protein content during fermentation. In addition, increasing non‐proteinaceous substances like chitin which is rich in the fungal cell walls and nucleic acids on account of fungal proliferation could also help to boost the protein content 107 .…”

Section: Beneficial Effects Of a Nigermentioning

confidence: 99%

“…Likewise, several positive reports on genetic engineering of A. niger have recently been reported. 103,171 The great potential of genome editing and engineering should also be exploited to reduce the challenges related to limiting the utilization of A. niger in the future, which will help to sustain the shrimp aquaculture with increased economic benefits.…”

Section: Genetic Engineeringmentioning

confidence: 99%

Beneficial effects, challenges and opportunities of the filamentous fungus,Aspergillus nigerwith special reference to the shrimp feed industry—A review

Jannathulla

Dayal

2022

Reviews in Aquaculture

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The shrimp industry, including its culture and feed sectors, exhibited steady growth over the years with a reduced cost–benefit ratio due to increasing production costs attributed mainly to enhanced feed costs. The unsustainable availability and rapid increase in fishmeal cost have forced the aquafeed sector to reduce dietary inclusion levels. The filamentous fungus, Aspergillus niger plays an important role in utilising plant protein sources for formulating cost‐effective shrimp feed. A. niger has been reported to produce numerous hydrolytic enzymes to degrade the limiting factors of plant proteins such as antinutrients and fibre fractions. This fungus enhanced not only the quantity but also the quality of proteins and amino acids, especially methionine and lysine. Likewise, it also increased the availability of soluble minerals, particularly phosphorous, by degrading the phytic‐phosphorus, thereby helping to replace dietary fishmeal up to 80% in penaeid shrimps. Consequently, this fungus could reduce feed formulation cost significantly. However, the mycotoxins produced by this fungus pose difficulties during large‐scale fermentation, and its implications on the workers and environment cast doubt about its efficacy and utility. However, recent developments indicate that A. niger could be an alternative choice in reducing the pressure on fishmeal demand by enhancing the nutrient utilization of plant‐based materials leading to the formulation of, cost‐effective feeds that can sustain shrimp culture and production in the future.

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“…Aspergillus niger is a powerful cell factory and among others, is efficiently used to produce organic acids such as citric-, oxalic-and gluconic acid (2,3). Citric-and oxalic acid are intracellularly produced and actively secreted (4)(5)(6)(7). In contrast, gluconic acid is the product of an extracellular conversion of glucose in a collaboration of three extracellular enzymes: glucose oxidase (GOx), lactonase and catalase (8).…”

mentioning

confidence: 99%

“…FIG6 Expression analysis of A. niger wildtype and AQP knock-out strains under hydrogen peroxide inducing conditions. Results of strain N402 (A), ΔaqpD (B), ΔaqpE (C) and ΔaqpF (D).…”

mentioning

confidence: 99%

Identification of Aspergillus niger aquaporins involved in hydrogen peroxide signaling

Laothanachareon

Asin-Garcia

Volkers

et al. 2022

Preprint

Self Cite

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Aspergillus niger is a robust microbial cell factory for organic acids production. However, the regulation of many industrially important pathways is still poorly understood. Recently the regulation of glucose oxidase (GOx) expression system involved in the biosynthesis of gluconic acid has been uncovered. Hydrogen peroxide, a by-product of the enzymatic conversion of glucose to gluconate, has a pivotal role as a signaling molecule in the induction of this system. In this study, facilitated diffusion of hydrogen peroxide via aquaporin water channels (AQPs) was studied. AQPs are transmembrane proteins of the major intrinsic proteins (MIPs) superfamily. In addition to water and glycerol, they may also transport small solutes such as hydrogen peroxide. Accordingly, the genome sequence of A. niger N402 was screened for putative AQPs. Seven were found which could be classified in three main groups. Their ability to facilitate diffusion of hydrogen peroxide was identified using yeast phenotypic growth assays and by studying AQP gene knock-outs in A. niger. Complementing from both yeast and A. niger experiments, the X-intrinsic protein AQPF plays apparent roles in facilitating hydrogen peroxide transport across cellular membrane.IMPORTANCEAquaporins (AQP) are integral membrane proteins that facilitate transfer of water, small solutes and hydrogen peroxide across the plasma membrane. As they have conserved structural features, it is not difficult to detect their presence in fungal genomes. Their specific roles in fungi are however less understood. Previously we have elucidated the role of the hydrogen peroxide as a second messenger in the induction and regulation of the expression of glucose oxidase involved in the extracellular conversion of glucose in gluconate (1). Here we identified the involvement of AQPF, a member of the X-intrinsic protein class in facilitating transport of the hydrogen peroxide signal across the cell membrane.

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Global Transcriptional Response of Aspergillus niger to Blocked Active Citrate Export through Deletion of the Exporter Gene

Cited by 7 publications

References 52 publications

Evaluation of Aspergillus niger Six Constitutive Strong Promoters by Fluorescent-Auxotrophic Selection Coupled with Flow Cytometry: A Case for Citric Acid Production

Evaluation of Aspergillus niger Six Constitutive Strong Promoters by Fluorescent-Auxotrophic Selection Coupled with Flow Cytometry: A Case for Citric Acid Production

Beneficial effects, challenges and opportunities of the filamentous fungus,Aspergillus nigerwith special reference to the shrimp feed industry—A review

Identification of Aspergillus niger aquaporins involved in hydrogen peroxide signaling

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