Project description:A. terreus LYT10 is an industrial strain for itaconic acid production, in which the biosynthesis of itaconic acid and glucose conversion rate were affected by temperature and initial concentration of itaconic acid in industrial production. RNA-seq was used to identify the key regulators related to tolerance mechanism toward various stress conditions.
Project description:A. terreus LYT10 is an industrial strain for itaconic acid production, in which the biosynthesis of itaconic acid and glucose conversion rate were affected by temperature and initial concentration of itaconic acid in industrial production. RNA-seq was used to identify the key regulators related to tolerance mechanism toward various stress conditions. A total of 4 samples were analzyed. The mycelia cultivated in itaconic acid production medium (IPM) on a rotary shaker at 220 rpm and 37°C for 36 h was considered as a reference. IPMs with 5 g/L and 40 g/L itaconic acid (pH 3.25) were used for high-acid culture respectively. High-temperature condition was possesed at 42°C for 36 h in IPM.
Project description:Aspergillus terreus is an emerging fungal pathogen in immunocompromised patients. Due to intrinsic resistance of AmB against A. terreus and acquiring resistance to azoles, alternative antifungal strategy needs investigation. Thus, we explored the activity of phytochemicals such as Shikonin, gallic acid, coumaric acid and quercetin against A. terreus. Amongst these, shikonin showed significant inhibition at MIC50;2 µg/ml, considered for proteome profiling.
Project description:Aim: We aimed to study the differential gene expressions of the target genes encoding H+-ATPase, the mitochondrial respiratory chain complex, and the alternative oxidase (AOX) in Aspergillus terreus NRRL1960 cultivated under low dissolved oxygen Methods:The fermentation samples were collected for RNA extraction.The barcoded RNA libraries were prepared using Lexogen’s Quant-Seq 3’ mRNA seq kit (Ion Torrent, Lexogen, Vienna, Austria). DNA templates for sequencing were prepared from 200 bp v3 OT2 kit and the Ion One Touch 2 platform. Sequencing was performed on the Ion Proton by Ion Torrent Suite v5.0.4. Raw sequences form each sample were uploaded and the dapter sequences were trimmed. The reads were aligned to A. terreus NIH2624 genome references downloaded from http://fungi.ensembl.org/info/website/ftp/index.html using Star 2.4.1d. The bam index files were generated. The quantification of gene expression was performed by Htseq v0.6.0 with -f and -s options indicating bam inputs and un-stranded reads, respectively. Cuffdiff v.2.1.1 was used to estimate the transcript abundance with the -no-diff and default options to generate the differential analysis for each described comparison. Results:From the gene expression (FPKM) results, genes in Cytochrome C complex and aoxA had the high expression level compared to pma encoding H+-ATPase. However, the different gene expression analysis results shown that the target genes in the mitochondrial respiratory chain complex were predominantly down regulated as observed from the log2(fold_change) less than -1 and and the gene encoding AOX (aoxA) were down regulated. Conclusions: In this study, the transcriptomic analysis were performed for better understanding the biosynthesis pathway and the regulation of target genes in itaconic acid cluster and the ATP regeneration pathway.
Project description:The filamentous fungus Aspergillus terreus is known to produce both industrially and pharmaceutically important secondary metabolites. The objective of this study is to investigate the effect of exogenously added butyrolactone I (BI) on the submerged culture of A. terreus, especially on the possible regulation of the secondary metabolism on the transcriptional level. In order to elucidate the presumed regulative role of butyrolactone I, a large-scale microarray gene expression study was designed and conducted with an industrially utilised A. terreus strain MUCL38669. A. terreus MUCL38669 was cultured in secondary metabolism inducing submerged conditions for nine days, where butyrolactone I was added at the beginning of the growth phase (at 24 hours p.i.), in the middle of the growth phase (at 96 hours p.i.) or in the late growth phase (at 120 hours p.i.), in addition to the control culture where no exogenous butyrolactone I was added. To obtain comprehensive gene expression profiles over the whole culture time, samples were taken at six time points: 24 hours, 48 hours, 96 hours, 120 hours, 144 hours and 216 hours post inoculation.
Project description:Aspergillus terreus has been described as an allergenic fungus in addition to causing infection to both human beings and plants. However, the allergens in this fungus are still unknown. The identification of allergens is essential to develop diagnostic and therapeutic avenues. Employing a proteomic approach, sixteen allergens were identified based on two-dimensional immuno-blotting with A. terreus susceptible patient sera. Amongst them, triosephosphate isomerase (Asp t 36), one of the dominant immunoglobulin E (IgE)-reactive proteins, was purified under native conditions. To characterize Asp t 36, the gene was cloned and expressed in E. coli. Phylogenetic analysis showed it to be highly conserved with close similarity to Dermatophagoides farinae. Four immuno-dominant epitopes were identified using synthetic peptides and mapped on the tertiary structure of Asp t 36. Among these, two were found to create a continuous surface patch on the 3D structure rendering it to be an IgE binding hotspot. Asp t 36 showed similar secondary structure and temperature sensitivity with other reported triosephosphate isomerase allergens. In-vivo studies using the murine model demonstrated rAsp t 36 able to stimulate airway inflammation, such as an influx of eosinophils, goblet cell hyperplasia, elevated serum immunoglobulins and induction of Th2 cytokines. Collectively, our results reveal the immunogenic property of Asp t 36, a major allergen from A. terreus for the first time in any fungi. This allergen could serve as a potent candidate for investigating component resolved diagnosis and immunotherapy.
Project description:Immunoresponsive gene 1 (IRG1) is one of the highest induced genes in macrophages under pro-inflammatory conditions and its function has been recently described: it codes for immune-responsive gene 1 protein/cis-aconitic acid decarboxylase (IRG1/CAD), an enzyme catalyzing the production of itaconic acid from cis-aconitic acid, a tricarboxylic acid (TCA) cycle intermediate. Itaconic acid possesses specific antimicrobial properties inhibiting isocitrate lyase, the first enzyme of the glyoxylate shunt, an anaplerotic pathway that bypasses the TCA cycle and enables bacteria to survive on limited carbon conditions. To elucidate the mechanisms underlying itaconic acid production through IRG1 induction in macrophages, we examined the transcriptional regulation of IRG1. Using a combination of literature information, transcription factor prediction models and genome-wide expression arrays, we inferred the regulatory network of IRG1 in mouse and human macrophages. 3 unstimulated (Control) and 3 LPS-stimulated human PBMC-derived macrophages
Project description:Immunoresponsive gene 1 (IRG1) is one of the highest induced genes in macrophages under pro-inflammatory conditions and its function has been recently described: it codes for immune-responsive gene 1 protein/cis-aconitic acid decarboxylase (IRG1/CAD), an enzyme catalyzing the production of itaconic acid from cis-aconitic acid, a tricarboxylic acid (TCA) cycle intermediate. Itaconic acid possesses specific antimicrobial properties inhibiting isocitrate lyase, the first enzyme of the glyoxylate shunt, an anaplerotic pathway that bypasses the TCA cycle and enables bacteria to survive on limited carbon conditions. To elucidate the mechanisms underlying itaconic acid production through IRG1 induction in macrophages, we examined the transcriptional regulation of IRG1. Using a combination of literature information, transcription factor prediction models and genome-wide expression arrays, we inferred the regulatory network of IRG1 in mouse and human macrophages. 3 unstimulated (Control) and 3 LPS-stimulated RAW 264.7 macrophages