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 (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 | Comment | Organism | Structure |
---|---|---|---|
Ag+ | 18.5% inhibition at 1 mM, 62.5% 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; complete inhibition at 1-5 mM | Thermochaetoides thermophila | |
Fe3+ | 5 mM, 70.3% loss of activity, substrate: carboxymethyl cellulose; 5 mM, 78.5% loss of activity, substrate: xylan; 8.3% inhibition at 1 mM, 78.5% inhibition at 5 mM | Thermochaetoides thermophila | |
Mg2+ | 22.7% inhibition at 1 mM, 50.6% inhibition at 5 mM; 5 mM, 51% loss of activity, substrate: xylan; 5 mM, 64.5% loss of activity, substrate: carboxymethyl cellulose | Thermochaetoides thermophila | |
Zn2+ | 17.1% inhibition at 1 mM, 55.2% at 5 mM; 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 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 10.28 mg/ml for beta-1,4-D-xylan | Thermochaetoides thermophila |
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 | |
Ca2+ | activates 19.5% at 5 mM | Thermochaetoides thermophila | |
Co2+ | 5 mM, 1.4fold activation, substrate: carboxymethyl cellulose | 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 47.5% at 1 mM, 38.6% at 5 mM | Thermochaetoides thermophila | |
additional information | poor effects by 1-5 mM Co2+ | Thermochaetoides thermophila |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
47000 | - |
calculated from sequence | Thermochaetoides thermophila |
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 | 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 (Comment) | Organism |
---|---|
- |
Thermochaetoides thermophila |
recombinant His-tagged enzyme from Pichia pastoris strain GS115 by nickel affinity chromatography | Thermochaetoides thermophila |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
3.05 | - |
purified recombinant His-tagged enzyme, pH 5.0, 55°C, substrate beta-xylan | Thermochaetoides thermophila |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
beta-1,4-D-glucan + H2O | bifunctional endoglucanase/xylanase enzyme | Thermochaetoides thermophila | ? | - |
? | |
beta-1,4-D-glucan + H2O | bifunctional endoglucanase/xylanase enzyme | Thermochaetoides thermophila IMI 039719 | ? | - |
? | |
beta-1,4-D-glucan + H2O | bifunctional endoglucanase/xylanase enzyme | Thermochaetoides thermophila DSM 1495 | ? | - |
? | |
beta-1,4-D-glucan + H2O | bifunctional endoglucanase/xylanase enzyme | Thermochaetoides thermophila CBS 144.50 | ? | - |
? | |
beta-1,4-D-xylan + H2O | 0.5% w/v xylan from beechwood | Thermochaetoides thermophila | ? | - |
? | |
beta-1,4-D-xylan + H2O | bifunctional endoglucanase/xylanase enzyme | Thermochaetoides thermophila | ? | - |
? | |
beta-1,4-D-xylan + H2O | 0.5% w/v xylan from beechwood | Thermochaetoides thermophila IMI 039719 | ? | - |
? | |
beta-1,4-D-xylan + H2O | bifunctional endoglucanase/xylanase enzyme | Thermochaetoides thermophila IMI 039719 | ? | - |
? | |
beta-1,4-D-xylan + H2O | 0.5% w/v xylan from beechwood | Thermochaetoides thermophila DSM 1495 | ? | - |
? | |
beta-1,4-D-xylan + H2O | bifunctional endoglucanase/xylanase enzyme | Thermochaetoides thermophila DSM 1495 | ? | - |
? | |
beta-1,4-D-xylan + H2O | 0.5% w/v xylan from beechwood | Thermochaetoides thermophila CBS 144.50 | ? | - |
? | |
beta-1,4-D-xylan + H2O | bifunctional endoglucanase/xylanase enzyme | Thermochaetoides thermophila CBS 144.50 | ? | - |
? | |
carboxymethyl cellulose + H2O | bifunctional endoglucanase/xylanase enzyme | Thermochaetoides thermophila | ? | - |
? | |
carboxymethyl cellulose + H2O | bifunctional endoglucanase/xylanase enzyme | Thermochaetoides thermophila IMI 039719 | ? | - |
? | |
carboxymethyl cellulose + H2O | bifunctional endoglucanase/xylanase enzyme | Thermochaetoides thermophila DSM 1495 | ? | - |
? | |
carboxymethyl cellulose + H2O | bifunctional endoglucanase/xylanase enzyme | Thermochaetoides thermophila CBS 144.50 | ? | - |
? | |
additional information | the bifunctional enzyme also shows endoglucanase activity (EC 3.2.1.4) hydrolyzing carboxymethyl cellulose and barley beta-glucan. Substrate specificity, overview. The enzyme shows endoglucanase, but not xylanase, 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. No xylanase activity with (+)-arabinogalactan, D-galacto-D-mannan, amylose, chitin, and sucrose | Thermochaetoides thermophila | ? | - |
? | |
additional information | the bifunctional enzyme also shows endoglucanase activity (EC 3.2.1.4) hydrolyzing carboxymethyl cellulose and barley beta-glucan. Substrate specificity, overview. The enzyme shows endoglucanase, but not xylanase, 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. No xylanase activity with (+)-arabinogalactan, D-galacto-D-mannan, amylose, chitin, and sucrose | Thermochaetoides thermophila IMI 039719 | ? | - |
? | |
additional information | the bifunctional enzyme also shows endoglucanase activity (EC 3.2.1.4) hydrolyzing carboxymethyl cellulose and barley beta-glucan. Substrate specificity, overview. The enzyme shows endoglucanase, but not xylanase, 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. No xylanase activity with (+)-arabinogalactan, D-galacto-D-mannan, amylose, chitin, and sucrose | Thermochaetoides thermophila DSM 1495 | ? | - |
? | |
additional information | the bifunctional enzyme also shows endoglucanase activity (EC 3.2.1.4) hydrolyzing carboxymethyl cellulose and barley beta-glucan. Substrate specificity, overview. The enzyme shows endoglucanase, but not xylanase, 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. No xylanase activity with (+)-arabinogalactan, D-galacto-D-mannan, amylose, chitin, and sucrose | Thermochaetoides thermophila CBS 144.50 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 47000, about, sequence calculation, x * 48000, recombinant His6-tagged enzyme, SDS-PAGE | Thermochaetoides thermophila |
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 3.2.1.4 | Thermochaetoides thermophila |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
- |
Thermochaetoides thermophila |
55 | - |
for CTendo7's xylanase activity and endoglucanase activity | Thermochaetoides thermophila |
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 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, half-life of endoxylanase activity is 100 min, 40% 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, half-life of endoxylanase activity is 40 min, 10% activity remaining after 80 min, inactivation 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, inactivation after 80 min | Thermochaetoides thermophila |
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 | |
0.00158 | - |
beta-1,4-D-xylan | pH 5.0, 55°C | Thermochaetoides thermophila | |
0.00158 | - |
beta-1,4-D-xylan | recombinant His-tagged enzyme, pH 5.0, 55°C | Thermochaetoides thermophila | |
0.00211 | - |
carboxymethyl cellulose | pH 5.0, 55°C | Thermochaetoides thermophila |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
- |
Thermochaetoides thermophila |
5 | - |
for CTendo7's xylanase activity and endoglucanase activity | Thermochaetoides thermophila |
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 |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Thermochaetoides thermophila | sequence calculation | - |
6.02 |
Thermochaetoides thermophila | calculated from sequence | - |
6.02 |
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 |