Effect of metal ion on itaconic acid fermentability for Aspergillus terreus
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ABSTRACT: Zn (Ⅱ) and Fe (Ⅱ) are the essential metal elements for the growth of microorganisms. It illustrated that more itaconic acid was achieved for 0.25 g/L ZnSO4·2H2O and 0.40 g/L FeSO4·2H2O than the control with 0.15 g/L ZnSO4·2H2O and 0.16 g/L FeSO4·2H2O after single factor assays for Aspergillus terreus.We furhter carried out transcriptome assays to uncover molecular mechanism of the enhanced itaconic acid fermentability for A. terreus with metal ion.Therefore, our study would provide a reference metal ion concentration ratio for itaconic and other biochemicals production.
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:N. oceanica IMET1 was inoculated into modified f/2 liquid medium, which was prepared with 35 g L−1 sea salt, 1 g L−1 NaNO3, 67 mg L−1 NaH2PO4*H2O, 3.65 mg L−1 FeCl3*6H2O, 4.37 mg L−1 Na2EDTA*2H2O, trace metal mix (0.0196 mg L−1 CuSO4*5H2O, 0.0126 mg L−1 NaMoO4*2H2O, 0.044 mg L−1 ZnSO4*7H2O, 0.01 mg L−1 CoCl2, and 0.36 mg L−1 MnCl2*4H2O), and vitamin mix (2.5 µg L−1 VB12, 2.5 µg L−1 biotin, and 0.5 µg L−1 thiamine HCl). The algal cells were grown in liquid cultures under continuous light (∼500 µmol photons m−2 s−1) at 25°C and aerated by bubbling with a mixture of 1.5% CO2 in air.
Project description:Methylorubrum extorquens AM1 is engineered to produce itaconic acid by heterologous expression of cis-aconitic acid decarboxylase. Mutation was also performed on phaR in Methylorubrum extorquens AM1, which regulate poly-beta-hydroxybutyrate accumulation, in attempt to increase carbon flux toward itaconic acid production. However, in our case, itaconic acid production by phaR mutant strain was not higher than that of the wildtype. Transcriptomic analysis was utilized in order to examine the cause for this phenomenon. RNA-seq analysis revealed that phaR mutation in the itaconic acid-producing strain might result in a complex regulatory rewiring at the gene expression level, which could cause a reduced resource flux toward ITA production. Also, RNA profiling gave a hint at the broad regulatory role of PhaR.
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:Acetate is a cost-effective and sustainable carbon source that, despite its potential, remains underutilized. This study employed biosensor-assisted adaptive laboratory evolution (ALE) to enhance itaconic acid production and acetate metabolism in Escherichia coli. The evolved E. coli W strains exhibited 65% increase in itaconic acid production and 71% increase in growth rate, and 45% increase in itaconic acid yield. A common 31-kb genomic deletion was identified in the evolved strains, with two genes, ecw_m2276 and ecw_m2277, driving the observed phenotypic changes. The evolved strains exhibited an intensified stringent response, which enhanced the acetate-utilizing pathway and resulted in over a 5,000% increase in the expression of the glyoxylate shunt, thereby boosting microbial growth. Overexpression of relA further replicated these enhanced phenotypes. Our findings highlight not only significant physiological improvements but also present a novel strategy for enhancing microbial growth and bioproduction from acetate, offering valuable insights for industrial biotechnology applications.
Project description:WT and ISG15 KO macrophages were stimulated with interferon-a and treated with itaconic acid and 4-octyl itaconate as they are known for anti-inflammatory effects. In this experiment, we further explored how itaconic acid and 4-octyl itaconate can play its role keeping ruxolitinib as a positive control in order to inhibit JAK/STAT pathway
Project description:Zinc and magnesium were the essectial activators for bioenzyme. Here, we assayed single factor assays for the effect of zinc and magnesium for Monascus purpureus.Then, we carried out transcriptome assays to uncover molecular mechanism of the enhanced fermentability of bioethanol and pigments for M. purpureus under the optimal metal ion concentration.This work would provide the fermentation process for biofuels and biochemicals.
Project description:Expression of target genes in itaconic acid cluster and electron transport chain in Aspergillus terreus NRRL1960 cultivated under low dissolved oxygen