Application | Comment | Organism |
---|---|---|
agriculture | 1,3-1,4-beta-D-glucanase are widely used as a feed additive to help non-ruminant animals digest plant fibers, with potential in increasing nutrition turnover rate and reducing sanitary problems | Fibrobacter succinogenes |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli and Pichia pastoris | Fibrobacter succinogenes |
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), heterologous expression of extracellular wild-type and mutant enzymes in Pichia pastoris | Fibrobacter succinogenes |
Crystallization (Comment) | Organism |
---|---|
apo-form and substrate complex structures of mutant V18Y/W203Y, to 1.53 A resolution | Fibrobacter succinogenes |
purified recombinant mutant V18Y/W203Y alone and in complex with product cellotetraose , from 0.1 M TrisHCl, pH 7.5, 0.3 M calcium acetate, and 29% PEG 5000 MME for the free mutant, and 0.15 M Tris, pH 8.5, 0.4 M calcium acetate and 33% PEG 5000 MME plus soaking in mother liquor with 5 mM cellotetraose for 1 h for the complexed mutant, X-ray diffraction structure determination and analysis at 1.53 A resolution, molecular replacement method | Fibrobacter succinogenes |
Protein Variants | Comment | Organism |
---|---|---|
233Stop | 28.4% of wild-type activity | Fibrobacter succinogenes |
M27D/M39R | 0.1% of wild-type activity | Fibrobacter succinogenes |
M27D/M39R | site-directed mutagenesis, almost inactive mutant | Fibrobacter succinogenes |
M27R/M39D | 0.2% of wild-type activity | Fibrobacter succinogenes |
M27R/M39D | site-directed mutagenesis, almost inactive mutant | Fibrobacter succinogenes |
M39F | 92.8% of wild-type activity | Fibrobacter succinogenes |
M39F | site-directed mutagenesis, 7% reduced activity compared to wild-type | Fibrobacter succinogenes |
additional information | construction of mutants based on catalytic domain, residues 1-243, with higher thermostability and specific activity | Fibrobacter succinogenes |
additional information | kinetic and thermostability analysis of the mutant enzymes, overview | Fibrobacter succinogenes |
S71F | loss of activity | Fibrobacter succinogenes |
S71F | site-directed mutagenesis, inactive mutant | Fibrobacter succinogenes |
S84D | 80.9% of wild-type activity | Fibrobacter succinogenes |
S84D | site-directed mutagenesis, 19% reduced activity compared to wild-type | Fibrobacter succinogenes |
V18Y | 104% of wild-type activity. Increase in thermostability by 2 degrees | Fibrobacter succinogenes |
V18Y | site-directed mutagenesis, 4% reduced activity compared to wild-type | Fibrobacter succinogenes |
V18Y/W203Y | 134.3% of wild-type activity | Fibrobacter succinogenes |
V18Y/W203Y | site-directed mutagenesis, 34% increased activity compared to wild-type | Fibrobacter succinogenes |
V61F | 20.9% of wild-type activity | Fibrobacter succinogenes |
V61F | site-directed mutagenesis, 79% reduced activity compared to wild-type | Fibrobacter succinogenes |
W203F | 87.6% of wild-type activity | Fibrobacter succinogenes |
W203F | site-directed mutagenesis, 13% reduced activity compared to wild-type | Fibrobacter succinogenes |
W203Y | 130.2% of wild-type activity | Fibrobacter succinogenes |
W203Y | site-directed mutagenesis, 30% increased activity compared to wild-type | Fibrobacter succinogenes |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | 2.25 mg/ml for wild-type enzyme, 2.44 mg/ml for mutant V18Y, 3.67 mg/ml for mutant W203Y, and 3.29 mg/ml for mutant V18Y/W203Y, with lichenan, pH 5.0, 50°C, calculated as reducing sugar released from substrate | Fibrobacter succinogenes |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Fibrobacter succinogenes | P17989 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, tag cleavage and again nickel affinity chromatography, recombinant extracellular wild-type and mutants V18Y, W203Y, and V18Y/W203Y enzymes from Pichia pastoris cell culture supernatant by dialysis and anion exchange chromatography | Fibrobacter succinogenes |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
5694 | - |
wild-type, expressed in Escherichia coli, pH 5.0, 50°C | Fibrobacter succinogenes |
6520 | - |
mutant V18Y, expressed in Escherichia coli, pH 5.0, 50°C | Fibrobacter succinogenes |
9263 | - |
mutant W203Y, expressed in Escherichia coli, pH 5.0, 50°C | Fibrobacter succinogenes |
9967 | - |
mutant V18Y/W203Y, expressed in Escherichia coli, pH 5.0, 50°C | Fibrobacter succinogenes |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,3-1,4-beta-D-glucan + H2O | - |
Fibrobacter succinogenes | ? | - |
? | |
barley beta-glucan + H2O | - |
Fibrobacter succinogenes | ? | - |
? | |
lichenan + H2O | - |
Fibrobacter succinogenes | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
1,3-1,4-beta-D-glucanase | - |
Fibrobacter succinogenes |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
assay at | Fibrobacter succinogenes |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
57 | - |
Tm of wild-type enzyme | Fibrobacter succinogenes |
59 | - |
Tm of mutant V18Y | Fibrobacter succinogenes |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.25 | - |
1,3-1,4-beta-D-glucan | wild-type, expressed in Escherichia coli, pH 5.0, 50°C | Fibrobacter succinogenes | |
2.44 | - |
1,3-1,4-beta-D-glucan | mutant V18Y, expressed in Escherichia coli, pH 5.0, 50°C | Fibrobacter succinogenes | |
3.29 | - |
1,3-1,4-beta-D-glucan | mutant V18Y/W203Y, expressed in Escherichia coli, pH 5.0, 50°C | Fibrobacter succinogenes | |
3.67 | - |
1,3-1,4-beta-D-glucan | mutant W203Y, expressed in Escherichia coli, pH 5.0, 50°C | Fibrobacter succinogenes | |
2924 | - |
lichenan | pH 5.0, 50°C, wild-type enzyme, calculated as reducing sugar released from substrate | Fibrobacter succinogenes | |
3224 | - |
lichenan | pH 5.0, 50°C, mutant V18Y, calculated as reducing sugar released from substrate | Fibrobacter succinogenes | |
5003 | - |
lichenan | pH 5.0, 50°C, mutant W203Y, calculated as reducing sugar released from substrate | Fibrobacter succinogenes | |
5090 | - |
lichenan | pH 5.0, 50°C, mutant V18Y/W203Y, calculated as reducing sugar released from substrate | Fibrobacter succinogenes |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
assay at | Fibrobacter succinogenes |