6 research outputs found

    Additional file 1: Figure S1. of Biochemical characterization of the xylan hydrolysis profile of the extracellular endo-xylanase from Geobacillus thermodenitrificans T12

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    FPLC purification of GtXynA1. Protein fraction not bound to the nickel column eluted with the first 65┬ámL eluent. Bound GtXynA1 protein was removed from the nickel column using a imidazole gradient (yellow line) which increased from 0┬ámM to 500┬ámM over a time span of 20┬ámin (A). Fractions 22ÔÇô25 were pooled and used for desalting the purified GtXynA1 (B). Fractions 8ÔÇô10 of the desalting column were pooled and then used for further experiments. (DOCX 311 kb

    Additional file 1: of Engineering Geobacillus thermodenitrificans to introduce cellulolytic activity; expression of native and heterologous cellulase genes

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    Table S1. Codon harmonization of celS from C. thermocellum. An overview of the codon harmonization method used to adapt the C. thermocellum derived exoglucanase encoding gene celS for expression in G. thermodenitrificans. (XLSX 78 kb

    Additional file 4: of Engineering Geobacillus thermodenitrificans to introduce cellulolytic activity; expression of native and heterologous cellulase genes

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    Figure S2. Congo red assays of cellulase expressing G. thermodenitrificans cultures. Congo red assays of G. thermodenitrificans cultures grown on LB2 medium with 1% carboxymethylcellulose. Each culture produces a different cellulase. ─é┬ś: empty plasmid (pNW33n) control; CelA: GH5 endoglucanase CelA (Geobacillus 70PC53); CelK: GH9 exoglucanase (C. thermocellum); GE40wt: GH5 endoglucanase (Geobacillus metagenome derived) containing its native signal peptide; GE40: GH5 endoglucanase (Geobacillus metagenome derived). (PDF 48 kb

    Additional file 2: of Engineering Geobacillus thermodenitrificans to introduce cellulolytic activity; expression of native and heterologous cellulase genes

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    Figure S1. Phylogenetic tree of GH5 family hydrolases. The evolutionary history was inferred by using the Maximum Likelihood method based on the JTT matrix-based model [42]. The percentage of trees in which the associated taxa clustered together is shown next to the branches. Initial tree(s) for the heuristic search were obtained automatically by applying Neighbor-Join and BioNJ algorithms to a matrix of pairwise distances estimated using the Jones Thornton Taylor (JTT) model, and then selecting the topology with superior log likelihood value. The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. The analysis involved 27 amino acid sequences. All positions containing gaps and missing data were eliminated. There were a total of 198 positions in the final dataset. Evolutionary analyses were conducted in MEGA7 [43]. Open circle: eukaryotic origin; closed triangle: thermophilic organism; closed diamond: sequences obtained in this study. (PDF 12 kb

    Additional file 2: Figure S2. of Biochemical characterization of the xylan hydrolysis profile of the extracellular endo-xylanase from Geobacillus thermodenitrificans T12

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    10% SDS-PAGE of purified endo-xylanase from G. thermodenitrificans T12 followed by PageBlue staining. Lane 1: Protein marker; Lane 2: Pellet fraction; Lane 3: cell-free extract; Lane 4: non-binding protein fraction from FPLC; Lane 5: Purified recombinant GtXynA1; Lane 6: Purified and desalted recombinant GtXynA1; Lane 7: Protein marker. (DOCX 614 kb
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