Journal: Biotechnology for Biofuels

Article Title: Mutation of the Xylanase regulator 1 causes a glucose blind hydrolase expressing phenotype in industrially used Trichoderma strains

doi: 10.1186/1754-6834-6-62

Figure Lengend Snippet: Analysis of transcript levels of MIHCs-encoding genes and xyr1 in T. reesei RutC30 and Iogen-M8. The xylanase overexpression strain Iogen-M8 ( A - E ) and its ancestor strain Rut-C30 ( F - J ) were precultured on glycerol and thereafter transferred to MA media without carbon source (NC) or containing 50 mM (w/v) glucose ( G ) or D-xylose (XO) at the indicated concentration, or 1.5 mM sophorose (SO) as an inducer. Samples were taken directly before transfer of mycelium (PRE), after 3 h of incubation (light grey bars), and after 6 h of incubation (dark grey bars). Transcription analyses of indicated genes from both strains were performed via qPCR using sar1 and act transcript levels for normalization. Results are given as relative transcript ratios in logarithmic scale (lg). The values provided in the figures are means from three biological experiments. Error bars indicate standard deviations. Transcript levels always refer to the reference sample (wild-type QM6a, NC 3 h).

Article Snippet: Sophorose induces cbh1 and cbh2 gene expression in Iogen-M4 (Figure C-E) as already observed before for Rut-C30 (Figure H-J) and, to a lesser extent, for Iogen-M8 (Figure C-E).

Techniques: Over Expression, Concentration Assay, Incubation

Journal: Biotechnology for Biofuels

Article Title: Mutation of the Xylanase regulator 1 causes a glucose blind hydrolase expressing phenotype in industrially used Trichoderma strains

doi: 10.1186/1754-6834-6-62

Figure Lengend Snippet: Analysis of transcript levels of MIHCs-encoding genes and xyr1 in T. reesei Iogen-M4. The parental strain of Iogen-M8, Iogen-M4, was precultured on glycerol and thereafter transferred to MA media without carbon source (NC) or containing 50 mM (w/v) glucose ( G ) or D-xylose (XO) at the indicated concentration, or 1.5 mM sophorose (SO) as an inducer. Samples were taken directly before transfer of mycelium (PRE), after 3 h of incubation (light grey bars), and after 6 h of incubation (dark grey bars). Transcription analyses of indicated genes ( A-E ) were performed via qPCR using sar1 and act transcript levels for normalization. Results are given as relative transcript ratios in logarithmic scale (lg). The values provided in the figures are means from three biological experiments. Error bars indicate standard deviations. Transcript levels always refer to the reference sample (wild-type QM6a, NC 3 h).

Article Snippet: Sophorose induces cbh1 and cbh2 gene expression in Iogen-M4 (Figure C-E) as already observed before for Rut-C30 (Figure H-J) and, to a lesser extent, for Iogen-M8 (Figure C-E).

Techniques: Concentration Assay, Incubation

Journal: Biotechnology for Biofuels

Article Title: Mutation of the Xylanase regulator 1 causes a glucose blind hydrolase expressing phenotype in industrially used Trichoderma strains

doi: 10.1186/1754-6834-6-62

Figure Lengend Snippet: Analysis of transcript levels of MIHCs-encoding genes and xyr1 in T. reesei Iogen-M4X. Iogen-M4X, which is Iogen-M4 with the mutated Xyr1, was precultured on glycerol and thereafter transferred to MA media without carbon source (NC) or containing 50 mM (w/v) glucose ( G ) or D-xylose (XO) at the indicated concentration, or 1.5 mM sophorose (SO) as an inducer. Samples were taken directly before transfer of mycelium (PRE), after 3 h of incubation (light grey bars), and after 6 h of incubation (dark grey bars). Transcription analyses of indicated genes ( A-E ) were performed via qPCR using sar1 and act transcript levels for normalization. Results are given as relative transcript ratios in logarithmic scale (lg). The values provided in the figures are means from three biological experiments. Error bars indicate standard deviations. Transcript levels always refer to the reference sample (wild-type QM6a, NC 3 h).

Article Snippet: Sophorose induces cbh1 and cbh2 gene expression in Iogen-M4 (Figure C-E) as already observed before for Rut-C30 (Figure H-J) and, to a lesser extent, for Iogen-M8 (Figure C-E).

Techniques: Concentration Assay, Incubation

Journal: Biotechnology for Biofuels

Article Title: Mutation of the Xylanase regulator 1 causes a glucose blind hydrolase expressing phenotype in industrially used Trichoderma strains

doi: 10.1186/1754-6834-6-62

Figure Lengend Snippet: Analysis of transcript levels of MIHCs-encoding genes and xyr1 in T. reesei QM6a. The wild-type QM6a was precultured on glycerol and thereafter transferred to MA media without carbon source (NC) or containing 50 mM (w/v) glucose ( G ) or D-xylose (XO) at the indicated concentration, or 1.5 mM sophorose (SO) as an inducer. Samples were taken directly before transfer of mycelium (PRE), after 3 h of incubation (light grey bars), and after 6 h of incubation (dark grey bars). Transcription analyses of indicated genes ( A-E ) were performed via qPCR using sar1 and act transcript levels for normalization. Results are given as relative transcript ratios in logarithmic scale (lg). The values provided in the figures are means from three biological experiments. Error bars indicate standard deviations. Transcript levels always refer to the reference sample (wild-type QM6a, NC 3 h). * indicates the reference sample, ND means not detected.

Article Snippet: Sophorose induces cbh1 and cbh2 gene expression in Iogen-M4 (Figure C-E) as already observed before for Rut-C30 (Figure H-J) and, to a lesser extent, for Iogen-M8 (Figure C-E).

Techniques: Concentration Assay, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Structural insights into glycoside hydrolase family 1 β-glucosidase: Selective oligosaccharide hydrolysis, synthesis, and product profiling

doi: 10.1016/j.jbc.2026.111293

Figure Lengend Snippet: The active site structures of ligand-bound Td2F2 E352Q. A , Td2F2 E352Q ( light beige ) in complex with sophorose ( lime ). B , Td2F2 E352Q ( gray ) in complex with laminaribiose ( cyan ). Water molecules are shown as red spheres . Direct and water-mediated hydrogen bonds are represented by magenta and light blue dashed lines , respectively. Distances between the carbon atoms of Arg325 and sophorose are shown as light orange dashed lines . The anomeric C1 atom of the nonreducing-end glucose, which is subject to the nucleophilic attack, is labeled. C , active site pocket of the sophorose complex. D , structural overlay of the sophorose and laminaribiose complexes. The black line indicates the distance between the C2 hydroxy groups of nonreducing end glucose. Numbers within the sugar rings correspond to subsite positions in the active site. Labels for the catalytic residues (nucleophile Glu352 mutated to Gln and acid/base catalyst Glu166) are underlined.

Article Snippet: Sucrose, glucose, cellobiose, and gentiobiose were purchased from Sigma-Aldrich (St Louis); sophorose, cellotriose, and cellotetraose were purchased from Megazyme; and laminaribiose was purchased from Carbosynth.

Techniques: Labeling

Journal: The Journal of Biological Chemistry

Article Title: Structural insights into glycoside hydrolase family 1 β-glucosidase: Selective oligosaccharide hydrolysis, synthesis, and product profiling

doi: 10.1016/j.jbc.2026.111293

Figure Lengend Snippet: MD simulations. A , C , E , and G , time evolution of RMSD for sugar ring atoms bound in the active site of three independent runs. B , D , F , and H , overlay of snapshot structures of the active site sampled every 50 ns in a selected run. A and B , the WT (Q352E) Td2F2 complexed with sophorose. C and D , the WT (Q352E) Td2F2 complexed with laminaribiose. E and F , the glucosyl–enzyme intermediate structure binding a glucose at subsite +1′. G and H , the glucosyl–enzyme intermediate structure binding a glucose at subsite +1.

Article Snippet: Sucrose, glucose, cellobiose, and gentiobiose were purchased from Sigma-Aldrich (St Louis); sophorose, cellotriose, and cellotetraose were purchased from Megazyme; and laminaribiose was purchased from Carbosynth.

Techniques: Binding Assay

Journal: The Journal of Biological Chemistry

Article Title: Structural insights into glycoside hydrolase family 1 β-glucosidase: Selective oligosaccharide hydrolysis, synthesis, and product profiling

doi: 10.1016/j.jbc.2026.111293

Figure Lengend Snippet: Schematic catalytic cycles of Td2F2 in hydrolysis and transglycosylation. A , hydrolysis : Sophorose can bind at subsites –1′/+1′ (waiting position, top panel) and then migrate to a catalytic pose at subsites −1/+1 ( middle panel). Cleavage of the glycosidic bond yields the release of the reducing-end glucose and the formation of the covalent glucosyl-enzyme intermediate, followed by hydrolysis of the intermediate to produce free glucose. B , transglycosylation : After cleavage of pNP-β-D-Glc (donor substrate) and release of pNP ( top panel), glucose (acceptor substrate) can bind in the alternative +1′ binding mode that orients the O2 hydroxy group to the anomeric carbon of the glucosyl-enzyme intermediate ( middle panel). Subsequently, glucose migrates to the catalytic +1 subsite, and sophorose is produced by transglycosylation ( bottom panel).

Article Snippet: Sucrose, glucose, cellobiose, and gentiobiose were purchased from Sigma-Aldrich (St Louis); sophorose, cellotriose, and cellotetraose were purchased from Megazyme; and laminaribiose was purchased from Carbosynth.

Techniques: Binding Assay, Produced

Journal: The Journal of Biological Chemistry

Article Title: Structural insights into glycoside hydrolase family 1 β-glucosidase: Selective oligosaccharide hydrolysis, synthesis, and product profiling

doi: 10.1016/j.jbc.2026.111293

Figure Lengend Snippet: Comparison of Td2F2 E352Q/sophorose with GH1 β-glucosidase/disaccharide complexes. Superimposition of the active site residues in the Td2F2 E352Q ( light beige )/sophorose ( lime ) complex with A , SGR_2426 ( hot pink )/sophorose ( gold ) (PDB code 9U98 ); B , Os3bglu7 ( cornflower blue )/laminaribiose ( tomato ) (PDB code 3AHT ); C , NkBgl ( orange )/cellobiose ( purple ) (PDB code 3VIK ); and D , BglB ( cyan )/thiocellobiose ( purple ) (PDB code 2O9R ). Numbers inside the sugar rings indicate the subsite positions of each sugar moiety. Labels for the catalytic residues are underlined .

Article Snippet: Sucrose, glucose, cellobiose, and gentiobiose were purchased from Sigma-Aldrich (St Louis); sophorose, cellotriose, and cellotetraose were purchased from Megazyme; and laminaribiose was purchased from Carbosynth.

Techniques: Comparison