Supplementary information files for An iron ore-based catalyst for producing hydrogen and metallurgical carbon via catalytic methane pyrolysis for decarbonisation of the steel industry

Supplementary files for article An iron ore-based catalyst for producing hydrogen and metallurgical carbon via catalytic methane pyrolysis for decarbonisation of the steel industry Experiments to investigate the catalytic pyrolysis of methane using an iron ore-based catalyst were carried out to optimize catalytic activity and examine the purity of the carbon produced from the process for the first time. Ball milling of the iron ore at 300 rpm for varying times – from 30 to 330 minutes – was studied to determine the effect of milling time on methane conversion. Optimal milling for 270 minutes led to a five-fold increase in methane conversion from ca. 1% to 5%. Further grinding resulted in a decline of methane conversion to 4% shown by SEM to correspond to an increase in particle size caused by agglomeration. Data from Raman and Mössbauer spectroscopy and H2 temperature programmed reduction indicated a change in phase from magnetite to maghemite and hematite (at the particle surface) as the grinding time increased. Analysis of the carbon produced as a byproduct of the reaction indicated a highly pure material with the potential to be used as an additive for steel production. </p

Differential localization of TMR-PAF26 in different <i>S. cerevisiae</i> strains.

<p>(A) Graph showing the percentage of cells that show TMR-PAF26 fluorescence limited to the cell envelope (class 1, white bars), in intracellular organelles (class 2, light grey bars), or filling whole cells (class 3, black bars), after treatment for 1 h with the peptide for each strain (as labeled on the x-axis). Cell location was determined by confocal microscopy. Results represent the means ± standard deviations of three experiments, each performed in triplicate. Dots above bars indicate statistical significance as compared to each class in the BY4741 parental strain control. Black dots indicate percentage lower than BY4741 and white dots higher. Two dots indicate p<0.01, and one dot p<0.05. (B) Representative confocal microscopy images are shown from the data set quantified in (A). Two examples of classes 1, 2 and 3 are labelled in strain BY4741. Bar: 10 µm.</p

Localization of fluorescently labeled PAF peptides in <i>N. crassa</i> cells.

<p>(A) Percentage of conidial population that do not show any TMR-peptide fluorescence signal (class “0”, striped bars), or show TMR-peptide fluorescence limited to cell envelopes (class 1, white bars), intracellular organelles (class 2, light grey bars), or filling the whole cytoplasm of the cells (class 3, black bars), after 1 h of treatment with 5 µM of each of the TMR-peptides. (B) Representative confocal images showing the localization of TMR-PAF26, TMR-PAF95 and TMR-PAF96 (in red) in conidia (left panels) and conidial germlings (right panels). For each peptide, the most common pattern of localization is shown (see A). Bars: 2 µm in conidia and 5 µm in germlings.</p

Activity and localization of PAF peptides in <i>A. fumigatus</i>.

<p>(A) Influence of PAF peptides on conidial germination. Percentage of germinated conidia was quantified by microscopy ∼5 h after treatment with the peptides at different concentrations. (B) Confocal microscopy of the localization of 5 µM TMR-labeled peptides (in red) in germlings of <i>A. fumigatus</i> expressing H1-GFP (in green) 1 h after treatment with the three peptides. Note that the left hand panels show brightfield images of germlings and the right panels show corresponding GFP-labeled nuclei in the same cells. The germlings were all treated for 1 h with the different PAF peptides before imaging. Note that the whole germlings treated with TMR-PAF26 are labeled by the fluorescent peptide, lack nuclei and are dead. This contrasts with both the TMR-PAF95 and TMR-PAF96 treated germlings that possess nuclei and are alive. The TMR-PAF95 treated germlings lack any labeling with the fluorescent peptide whilst TMR-PAF96 is bound only to the cell envelope of the germlings. Bar: 10 µm.</p

Saccharomyces cerevisiae gene deletion mutants used in this study.

<p>Saccharomyces cerevisiae gene deletion mutants used in this study.</p

Dose-response curves of the effects of the PAF peptides on conidial germination and viability of conidia of <i>N. crassa</i>.

<p>(A) Percentage of germinated conidia was quantified by light microscopy 3 h after treatment with the peptides. (B) Percentage of viable cells after 3 h of peptide treatment was quantified using the cell death marker propidium iodide. The peptides used were PAF26 (black circles), PAF95 (white triangles) and PAF96 (white squares).</p

Activity and localization of PAF peptides in yeast.

<p>(A) Serial 5-fold dilutions of exponentially growing <i>S. cerevisiae</i> BY4741 cells treated with 64 µM of each peptide for 24 h, and subsequently plated onto YPD peptide-free plates. (B) Brightfield (images on the left) and corresponding confocal microscopy (images on the right) showing the localization in yeast cells (1×10⁶ cells/ml) after treatment with 2.5 µM of each of the TMR-labeled peptides (in red) for 30 min. Bar: 10 µm.</p

Interaction and internalization of TMR-PAF26 in <i>A. fumigatus</i>.

<p>(A) Sequential images show the interaction and internalization of 5 µM TMR-PAF26 (in red) in actively growing conidial germlings of <i>A. fumigatus</i> expressing H1-GFP (in green) over a period of 25 min. (B) Detail of a germling present in (A) showing merged fluorescent and brightfield images. Note accumulation of TMR-PAF26 in vacuolar compartments (arrows) which undergo expansion and become more intensely fluorescent with time. This coincided with the initiation of the breakdown and loss of fluorescence of the nuclei (asterisks). (C) Detailed images of a germling undergoing these effects over a period of 5 min recorded 50 min after peptide addition. See also Movie S1 for the entire sequential time course of panel (A). Bar: 10 µm in (A) and 4 µm in (B) and (C).</p

Subcellular localization of PAF95 and PAF96 in conidial germlings of <i>N. crassa</i>.

<p>(A) Co-localization of TMR-PAF95 (in red) with the vacuolar marker cDFFDA (in green) after treatment of the germlings with 5 µM TMR-PAF95 for 1 h. Note co-localization within the vacuolar network in these merged images. (B) Localization of TMR-PAF96 (in red) and the cell wall stain CFW (in blue) after treatment with TMR-PAF96 for 1 h. Note that in regions of the cell envelope the calcofluor white staining is exterior to that of the TMR-PAF96 labelling (see inset) in the merged image. Bar: 5 µm.</p

Effect of T-720 on the oxalic acid (OA) production of wood decay basidiomycetes.

<p>Effect of T-720 on the oxalic acid (OA) production of wood decay basidiomycetes.</p