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Literature summary for 3.2.1.4 extracted from
- Characterization of a novel thermostable GH7 endoglucanase from Chaetomium thermophilum capable of xylan hydrolysis (2018), Int. J. Biol. Macromol., 117, 342-349 .
Application
| Application | Comment | Organism |
|---|---|---|
| industry | the enzyme is a promising candidate for industrial lignocellulosic biomass conversion. Generation of soluble oligosaccharides from lignocellulose is a critical step in bioethanol production. The enzyme produces cello-oligosaccharides and xylo-oligosaccharides from the continuous enzymatic saccharification of sodium carboxymethyl cellulose and xylan, respectively | Thermochaetoides thermophila |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene ctendo7, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of C-terminally His6-tagged enzyme in Pichia pastoris strain GS115 | Thermochaetoides thermophila |
| heterologously expressed in Pichia pastoris | Thermochaetoides thermophila |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Ag+ | 23% inhibition at 1 mM, 68% inhibition at 5 mM; 5 mM, 62.5% loss of activity, substrate: xylan; 5 mM, 68.25% loss of activity, substrate: carboxymethyl cellulose | Thermochaetoides thermophila |  |
| Co2+ | 5 mM, 2% loss of activity, substrate: xylan | Thermochaetoides thermophila | |
| Cu2+ | 5 mM, complete loss of activity, substrate: carboxymethyl cellulose; 5 mM, complete loss of activity, substrate: xylan; 75% inhibition at 1 mM, complete inhibition at 5 mM | Thermochaetoides thermophila | |
| Fe3+ | 14% inhibition at 1 mM, 70% inhibition at 5 mM; 5 mM, 70.3% loss of activity, substrate: carboxymethyl cellulose; 5 mM, 78.5% loss of activity, substrate: xylan | Thermochaetoides thermophila | |
| Mg2+ | 5 mM, 51% loss of activity, substrate: xylan; 5 mM, 64.5% loss of activity, substrate: carboxymethyl cellulose; 6% inhibition at 1 mM, 64.5% inhibition at 5 mM | Thermochaetoides thermophila | |
| Zn2+ | 5 mM, 4.4% loss of activity, substrate: carboxymethyl cellulose; 5 mM, 55% loss of activity, substrate: xylan | Thermochaetoides thermophila | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
beta-1,4-D-xylan | Km-value: 10.28 mg/ml, pH 5.0, 55°C | Thermochaetoides thermophila | |
| additional information | - |
carboxymethyl cellulose | Km-value: 79.19 mg/ml, pH 5.0, 55°C | Thermochaetoides thermophila | |
| additional information | - |
beta-1,4-D-glucan | Km-value: 9.82 mg/ml, pH 5.0, 55°C | Thermochaetoides thermophila | |
| additional information | - |
additional information | Michaelis-Menten kinetics, Km is 9.82 mg/ml for barley beta-1,4-D-glucan | Thermochaetoides thermophila |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | 5 mM, 1.1fold activation, substrate: carboxymethyl cellulose | Thermochaetoides thermophila | |
| Ca2+ | 5 mM, 1.2fold activation, substrate: xylan | Thermochaetoides thermophila | |
| Co2+ | 5 mM, 1.4fold activation, substrate: carboxymethyl cellulose | Thermochaetoides thermophila | |
| Co2+ | activates at 1-5 mM | Thermochaetoides thermophila | |
| Mn2+ | 5 mM, 1.4fold activation, substrate: xylan | Thermochaetoides thermophila | |
| Mn2+ | 5 mM, 1.7fold activation, substrate: carboxymethyl cellulose | Thermochaetoides thermophila | |
| Mn2+ | activates at 1-5 mM | Thermochaetoides thermophila | |
| additional information | poor effects by Ca2+ on endoglucanase activity at 1-5 mM | Thermochaetoides thermophila | |
| Zn2+ | activates at 1 mM, slight inhibition at 5 mM | Thermochaetoides thermophila | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 47000 | - |
calculated from sequence | Thermochaetoides thermophila |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Thermochaetoides thermophila | G0S9G3 | - |
- |
| Thermochaetoides thermophila CBS 144.50 | G0S9G3 | - |
- |
| Thermochaetoides thermophila DSM 1495 | G0S9G3 | - |
- |
| Thermochaetoides thermophila IMI 039719 | G0S9G3 | - |
- |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| glycoprotein | - |
Thermochaetoides thermophila |
| glycoprotein | predicted one N-linked glycosylation site (N261) and three O-linked glycosylation sites (T32, T312 and T332), respectively | Thermochaetoides thermophila |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Thermochaetoides thermophila |
| recombinant His-tagged enzyme from Pichia pastoris strain GS115 by nickel affinity chromatography | Thermochaetoides thermophila |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 2.22 | - |
purified recombinant His-tagged enzyme, pH 5.0, 55°C, substrate beta-glucan | Thermochaetoides thermophila |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| barley beta-1,4-D-glucan + H2O | best endoglucanase substrate | Thermochaetoides thermophila | ? | - |
? | |
| beta-1,4-D-glucan + H2O | bifunctional endoglucanase/xylanase enzyme | Thermochaetoides thermophila | ? | - |
? | |
| beta-1,4-D-xylan + H2O | bifunctional endoglucanase/xylanase enzyme | Thermochaetoides thermophila | ? | - |
? | |
| carboxymethyl cellulose + H2O | 1% w/v CMC-Na | Thermochaetoides thermophila | ? | - |
? | |
| carboxymethyl cellulose + H2O | bifunctional endoglucanase/xylanase enzyme | Thermochaetoides thermophila | ? | - |
? | |
| additional information | the bifunctional enzyme also shows endoxylanase activity (EC 3.2.1.8) hydrolyzing beta-1,4-D-xylan from beechwood. Substrate specificity, overview. No endoglucanase activity on (+)-arabinogalactan, D-galacto-D-mannan, amylose, chitin, and sucrose. The enzyme shows activity with pretreated wheat straw and filter paper. CTendo7 produces cellooligosaccharides and xylooligosaccharides from the continuous enzymatic saccharification of carboxymethyl cellulose-Na and xylan, respectively | Thermochaetoides thermophila | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 47000, about, sequence calculation, x * 48000, recombinant His6-tagged enzyme, SDS-PAGE | Thermochaetoides thermophila |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| bifunctional endoglucanase/xylanase | - |
Thermochaetoides thermophila |
| ctendo7 | - |
Thermochaetoides thermophila |
| CTHT_0045780 | gene name, UniProt | Thermochaetoides thermophila |
| GH7 endoglucanase | - |
Thermochaetoides thermophila |
| More | cf. EC 3.2.1.8 | Thermochaetoides thermophila |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 55 | - |
- |
Thermochaetoides thermophila |
| 55 | - |
for CTendo7's xylanase activity and endoglucanase activity | Thermochaetoides thermophila |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | 90 | activity range, profile overview | Thermochaetoides thermophila |
| 40 | 80 | 40°C: about 40% of maximal activity, 80°C: about 50% of maximal activity, substrate: xylan | Thermochaetoides thermophila |
| 40 | 80 | 40°C: about 50% of maximal activity, 80°C: about 50% of maximal activity, substrate: carboxymethyl cellulose | Thermochaetoides thermophila |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 60 | - |
120 min, enzyme retains about 85% of its initial activity, xylanase ativity | Thermochaetoides thermophila |
| 60 | - |
120 min, enzyme retains more than 80% of its initial activity, endoglucanase activity | Thermochaetoides thermophila |
| 60 | - |
purified recombinant His-tagged enzyme, 90% activity remaining after 120 min | Thermochaetoides thermophila |
| 70 | - |
60 min, enzyme retains 66.2% of its initial activity, xylanase ativity | Thermochaetoides thermophila |
| 70 | - |
60 min, enzyme retains 74.3% of its initial activity, endoglucanase activity | Thermochaetoides thermophila |
| 70 | - |
purified recombinant His-tagged enzyme, 74.3% remaining after 60 min, half-life of endoxylanase activity is 110 min, 30% activity remaining after 120 min | Thermochaetoides thermophila |
| 80 | - |
60 min, enzyme retains 61.3% of its initial activity, endoglucanase activity | Thermochaetoides thermophila |
| 80 | - |
60 min, enzyme retains about 50% of its initial activity, xylanase ativity | Thermochaetoides thermophila |
| 80 | - |
purified recombinant His-tagged enzyme, 61.3% activity remaining after 60 min, half-life of endoxylanase activity is 80 min, 18% activity remaining after 120 min | Thermochaetoides thermophila |
| 90 | - |
40 min, enzyme retains about 28% of its initial activity, endoglucanase activity | Thermochaetoides thermophila |
| 90 | - |
60 min, enzyme retains about 30% of its initial activity, xylanase ativity | Thermochaetoides thermophila |
| 90 | - |
purified recombinant His-tagged enzyme, half-life of endoxylanase activity is 30 min, 20% activity remaining after 60 min, after 120 min | Thermochaetoides thermophila |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0007 | - |
beta-1,4-D-glucan | pH 5.0, 55°C | Thermochaetoides thermophila | |
| 0.0007 | - |
barley beta-1,4-D-glucan | recombinant His-tagged enzyme, pH 5.0, 55°C | Thermochaetoides thermophila | |
| 0.00158 | - |
beta-1,4-D-xylan | pH 5.0, 55°C | Thermochaetoides thermophila | |
| 0.00211 | - |
carboxymethyl cellulose | pH 5.0, 55°C | Thermochaetoides thermophila | |
| 0.00211 | - |
carboxymethyl cellulose | recombinant His-tagged enzyme, pH 5.0, 55°C | Thermochaetoides thermophila |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 5 | - |
- |
Thermochaetoides thermophila |
| 5 | - |
for CTendo7's xylanase activity and endoglucanase activity | Thermochaetoides thermophila |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 3.5 | 9.5 | activity range, profile overview | Thermochaetoides thermophila |
| 4 | 7 | pH 4.0: about 75% of maximal activity, pH 7.0: about 35% of maximal activity, substrate: carboxymethyl cellulose | Thermochaetoides thermophila |
| 4 | 7 | pH 4.0: about 85% of maximal activity, pH 7.0: about 45% of maximal activity, substrate: xylan | Thermochaetoides thermophila |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Thermochaetoides thermophila | sequence calculation | - |
6.02 |
| Thermochaetoides thermophila | calculated from sequence | - |
6.02 |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | the enzyme belongs to the glycosyl hydrolase family 7, GH7. Residue Glu197 acts as the active-site nucleophile and Glu202 is the acid-base catalyst, and this is consistent with a catalytic mechanism leading to a net retention of configuration at the anomeric carbon in the deep substrate binding cleft | Thermochaetoides thermophila |
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