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Correlating citric acid formation and manganese limitation in Aspergillus niger using transcriptome and proteome analysis

ABSTRACT: Aspergillus niger is well known for its capability to produce citrate in high amounts but the detailed metabolic response causing citrate production has not been fully elucidated. Manganese is known to have an important effect as its limitation is a requirement to obtain high-level citrate formation. To identify the translational regulation causing citric acid overflow metabolism, transcriptome and proteome data from cultivations in manganese limitation and manganese excess conditions were analyzed. In addition to four already described main responses, two novel events were identified. The first metabolic response was down regulation of phosphoenolpyruvate carboxykinase (PEPCK) during manganese limited conditions, which was confirmed by in vivo experiments. Down regulation of the first step in the gluconeogenesis, while maintaining a high activity through glycolysis, promoted secretion of citrate into the medium as an alternative regulatory mechanism for adjusting the intracellular concentrations of TCA intermediates. The other novel observation was down regulation of two cation transporters at manganese limited conditions. It was hypothesized that lowered cation transport across the mitochondrial membrane reduced the ability of the cell to maintain homeostasis thereby favoring citric acid secretion. Finally, upregulation of an ABC transporter was measured, which was assumed to be a citrate permease. 9 samples in total. Three conditions in triplicates

ORGANISM(S): Aspergillus niger

SUBMITTER: Lars Poulsen

PROVIDER: E-GEOD-61985 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Correlating citric acid formation and manganese limitation in Aspergillus niger using transcriptome and proteome analysis

Project description:Aspergillus niger is well known for its capability to produce citrate in high amounts but the detailed metabolic response causing citrate production has not been fully elucidated. Manganese is known to have an important effect as its limitation is a requirement to obtain high-level citrate formation. To identify the translational regulation causing citric acid overflow metabolism, transcriptome and proteome data from cultivations in manganese limitation and manganese excess conditions were analyzed. In addition to four already described main responses, two novel events were identified. The first metabolic response was down regulation of phosphoenolpyruvate carboxykinase (PEPCK) during manganese limited conditions, which was confirmed by in vivo experiments. Down regulation of the first step in the gluconeogenesis, while maintaining a high activity through glycolysis, promoted secretion of citrate into the medium as an alternative regulatory mechanism for adjusting the intracellular concentrations of TCA intermediates. The other novel observation was down regulation of two cation transporters at manganese limited conditions. It was hypothesized that lowered cation transport across the mitochondrial membrane reduced the ability of the cell to maintain homeostasis thereby favoring citric acid secretion. Finally, upregulation of an ABC transporter was measured, which was assumed to be a citrate permease.

2014-10-03 | GSE61985 | GEO

Cap analysis gene expression (CAGE) of Aspergillus luchuensis mut. kawachii control and laeA disruptant strains

Project description:LaeA, a putative methyltransferase, is a global regulator for metabolic and development process in filamentous fungi. We characterized the laeA homologous genes in the white koji fungus, Aspergillus luchuensis mut. kawachii (A. kawachii) to examine their role in citric acid production. The ΔlaeA strain showed a significant reduction in the citric acid productivity. The cap-analysis gene expression (CAGE) revealed the laeA is required for the gene expression of a putative citrate exporter encoding cexA, which has a critical role for the citric acid production. The deficient citric acid productivity of the ΔlaeA strain was remedied by overexpression of cexA to a comparative level to that of the cexA overexpressing ΔcexA strain. In addition, chromatin immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) analysis indicated that LaeA regulates gene expression of the citrate exporter encoding cexA gene via histone H3K4 and histone H3K9 methylation levels. These results indicate that LaeA is involved in the citric acid production through epigenetic regulation of cexA in A. kawachii.

2019-08-15 | GSE135849 | GEO

Pseudomonas aerugiosa tctED deletion strain compared to wildtype

Project description:Purpose of study was to investigate whole genome expression changes of a strain with deletion of the two-component system TctD-TctE and determine genes dysregulate relative to the parental wildtype to gain insight into possible regulatory targets of TctD-TctE. TctD-TctE is a two-component system in Pseudomonas aeruginosa that responds to and regulates uptake of tricarboxylic acids such as citric acid. It accomplishes this through derepression of the porin encoding the gene opdH, thereby regulating influx of citrate metabolites from the surrounding environment. Deletion of the tctED operon (ΔtctED) resulted in a reduced growth phenotype when citric acid is present in media. In the ΔtctED strain the presence of citric acid was found to have an inhibitory effect on growth. When the alternative carbon source arginine was present, wildtype levels of growth could not be restored. Static cultures of ΔtctED had low cell density in the presence of citric acid but maintained the same levels of biofilm formation compared to conditions when no citric acid was present. This suggests a dysregulation of biofilm formation in the presence of citric acid. In the ΔtctED strain there was also greater accumulation of tobramycin within the biofilm compared to the PA14 wildtype strain. Additionally, analysis of whole-genome expression found that multiple metabolic genes were dysregulated in ΔtctED. Here it is concluded that TctD-TctE is involved in biofilm tolerance to tobramycin in the presence of citrate metabolites.

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Expanding the Pathways of Manganese Homeostasis: Role of a Small Manganese Chaperone Protein, MntS

Project description:Escherichia coli possesses >65 small proteins of <50 amino acids, many of which are uncharacterized. We have identified a new small protein, MntS, involved in manganese homeostasis. Manganese is a critical micronutrient, serving as an enzyme cofactor and protecting against oxidative stress. Yet manganese is toxic in excess and little is known about its function in cells. Bacteria carefully control intracellular manganese levels using the transcription regulator MntR. Before this work, mntH, which encodes a manganese importer, was the only gene known to respond to manganese via MntR repression in E. coli K12. We demonstrated that mntS is another member of the MntR manganese regulon. We also identified yebN, which encodes a putative manganese efflux pump, as the first gene positively regulated by MntR in Enterobacteria. Since MntS is expressed when manganese levels are low, causes manganese sensitivity when overexpressed, and binds manganese, we propose that MntS may be a manganese chaperone. This study reveals new factors involved in manganese regulation and metabolism and expands our knowledge of how small proteins function. Two E. coli strains, MG1655 (wild type) and GSO458 (Delta-mntR) were grown to OD600 ~ 0.5 in M9 glucose media at 37 M-BM-:C and treated with 10 microM MnCl2. In the first experiment, this incubation with 10 microM MnCl2 was for 60 min and in the second experiment, it was for 10 min. RNA was extracted using the hot phenol method and cDNA prepared and hybridized according the manufacturer's instructions (Affymetrix).

2011-12-08 | E-GEOD-25318 | biostudies-arrayexpress

Medium pH influences citric acid and lipid production in oleaginous yeast Yarrowia lipolytica

Project description:The goal of this study is to find out how Yarrowia lipolytica W29’s transcriptome changes in response to different pHs in certain medium, where the production of citric acid and lipid vary significantly at different pHs. The changes in transcriptome will give us insight about the mechanism of citric acid and/or lipid production are regulated. We harvested Y. lipolytica cells from bioreactor cultures from three different pHs: 2.0, 4.0 and 6.0 in triplicates. We sequenced the mRNA by Illumina HiSeq 2500 and generated an average of 20 million reads for each sample. We did not find distinct gene expression changes for the enzymes right downstream the intersection where citrate is cleaved into acetyl-CoA. Instead, from gene ontology enrichment analysis, we observed proteins with transport activities were overrepresented in gene groups that were up-regulated higher pH conditions, where citric acid secretion is favored.

2019-04-26 | GSE80717 | GEO

Gene expression profiling of SHSY5Y cells exposed to manganese

Project description:The trace element manganese is essential for normal development for all the organisms. Overexposure of manganese may leads to multiple neuronal disorders such as Parkinson, manganism. To explore the molecular mechanism of manganese induced neurotoxicity, gene expression profiling was performed on human neuroblastoma SHSY-5Y cells. Cells were exposed to sub-lethal concentration of manganese (100 Î¼M) for 24 hrs. Our result demonstrates that manganese alter multiple biological pathways including chromatin assembly, neurogenesis and apoptotic pathways.Cells grown in 75mm2 flask. three replicates for each sample SHSY5Y cells grown in MEM:F12 media (1:1) were treated with manganese and total RNA was isolated from cells after 24 hour exposure Three replicate were used for the experiment

2016-07-01 | E-GEOD-70845 | biostudies-arrayexpress

Expanding the Pathways of Manganese Homeostasis: Role of a Small Manganese Chaperone Protein, MntS

2011-12-08 | GSE25318 | GEO

Aspergillus niger

Project description:The manganese (II) ion dependent transcriptome of the citric acid overproducer fungus Aspergillus niger

| PRJNA1032880 | ENA

Aspergillus niger

Project description:Correlating citric acid formation and manganese limitation in Aspergillus niger using transcriptome and proteome analysis

| PRJNA262895 | ENA

MntR(Rv2788): a transcriptional regulator that controls manganese homeostasis in Mycobacterium tuberculosis.

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