Cloned (Comment) | Organism |
---|---|
12 mutant genes with changes in fice conserved residues of Cel9A-68 are constructed, cloned and expressed in Escherichia coli | Thermobifida fusca |
Protein Variants | Comment | Organism |
---|---|---|
D55A | activity with carboxymethyl cellulose is 0.2% of wild-type activity, activity with phosphoric acid-swollen cellulose is 1.2% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 1.6% of wild-type activity | Thermobifida fusca |
D55A/D58A | activity with carboxymethyl cellulose is less than 0.1% of wild-type activity, activity with phosphoric acid-swollen cellulose is 0.13% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 0.5% of wild-type activity | Thermobifida fusca |
D55N | activity with carboxymethyl cellulose is 0.3% of wild-type activity, activity with phosphoric acid-swollen cellulose is 1.6% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 2.2% of wild-type activity | Thermobifida fusca |
D58A | activity with carboxymethyl cellulose is 0.4% of wild-type activity, activity with phosphoric acid-swollen cellulose is 1.8% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 3.5% of wild-type activity, mutant enzyme loses about 90% of the initial activity after 15 h at 65°C, compared to 10% loss of wild-type activity | Thermobifida fusca |
D58N | activity with carboxymethyl cellulose is 0.45% of wild-type activity, activity with phosphoric acid-swollen cellulose is 2% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 2.7% of wild-type activity, mutant enzyme loses about 20% of the initial activity after 15 h at 65°C, compared to 10% loss of wild-type activity | Thermobifida fusca |
DELTAT245-L251/R252K | activity with carboxymethyl cellulose, acid-swollen cellulose or bacterial microcrystalline cellulose from Acetobacter xylinum is nearly identical to wild-type activity | Thermobifida fusca |
E424A | activity with carboxymethyl cellulose is 0.13% of wild-type activity, activity with phosphoric acid-swollen cellulose is 0.2% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 0.43% of wild-type activity, kcat/Km for 2,4-dinitrophenyl beta-D-cellobioside is 8.5fold higher than the wild-type value | Thermobifida fusca |
E424G | activity with carboxymethyl cellulose is 0.3% of wild-type activity, activity with phosphoric acid-swollen cellulose is 1.1% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 2.5% of wild-type activity, kcat/Km for 2,4-dinitrophenyl beta-D-cellobioside is 123.8fold higher than the wild-type value | Thermobifida fusca |
E424Q | activity with carboxymethyl cellulose is less than 0.1% of wild-type activity, activity with phosphoric acid-swollen cellulose is 0.15% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 1.1% of wild-type activity | Thermobifida fusca |
additional information | the FnIII-like domain is deleted from Cel19A-90, reducing activity with bacterial microcrystalline cellulose to 43% of the wild-type | Thermobifida fusca |
Y206F | activity with carboxymethyl cellulose is 7% of wild-type activity, activity with phosphoric acid-swollen cellulose is 4.1% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 8.3% of wild-type activity, mutant enzyme loses about 20% of the initial activity after 15 h at 65°C, compared to 10% loss of wild-type activity | Thermobifida fusca |
Y206S | activity with carboxymethyl cellulose is 0.5% of wild-type activity, activity with phosphoric acid-swollen cellulose is 1.3% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 1.8% of wild-type activity | Thermobifida fusca |
Y318A | activity with carboxymethyl cellulose is 5fold higher than wild-type activity, activity with phosphoric acid-swollen cellulose is 28% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 14.2% of wild-type activity, mutant enzyme loses about 30% of the initial activity after 15 h at 65°C, compared to 10% loss of wild-type activity | Thermobifida fusca |
Y318F | activity with carboxymethyl cellulose is 6.7fold higher than wild-type activity, activity with phosphoric acid-swollen cellulose is 75% of wild-type activity, activity with bacterial microcrystalline cellulose from Acetobacter xylinum is 16.6% of wild-type activity | Thermobifida fusca |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.197 | - |
2,4-dinitrophenyl beta-D-cellobioside | 50°C, mutant enzyme E424G | Thermobifida fusca | |
0.29 | - |
2,4-dinitrophenyl beta-D-cellobioside | 50°C, mutant enzyme E424A | Thermobifida fusca | |
0.697 | - |
2,4-dinitrophenyl beta-D-cellobioside | 50°C, wild-type enzyme Cel9A-68 | Thermobifida fusca |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermobifida fusca | P26221 | endoglucanase E-4 precursor; Cel9A | - |
Thermobifida fusca Cel9A | P26221 | endoglucanase E-4 precursor; Cel9A | - |
Purification (Comment) | Organism |
---|---|
- |
Thermobifida fusca |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,4-dinitrophenyl beta-D-cellobioside + H2O | - |
Thermobifida fusca | ? | - |
? | |
2,4-dinitrophenyl beta-D-cellobioside + H2O | - |
Thermobifida fusca Cel9A | ? | - |
? | |
bacterial microcrystalline cellulose + H2O | - |
Thermobifida fusca | ? | - |
? | |
bacterial microcrystalline cellulose + H2O | - |
Thermobifida fusca Cel9A | ? | - |
? | |
carboxymethyl cellulose + H2O | - |
Thermobifida fusca | ? | - |
? | |
carboxymethyl cellulose + H2O | - |
Thermobifida fusca Cel9A | ? | - |
? | |
phosphoric acid-swollen cellulose + H2O | - |
Thermobifida fusca | ? | - |
? | |
phosphoric acid-swollen cellulose + H2O | - |
Thermobifida fusca Cel9A | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Cel9A-68 | Cel9A is secreted as a 90400 Da protein. Limited proteolysis of Cel9A produces Cel9A-68 | Thermobifida fusca |
Cel9A-90 | Cel9A is secreted as a 90400 Da protein. Limited proteolysis of Cel9A produces Cel9A-68 | Thermobifida fusca |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
15 h, wild-type enzyme and mutant enzymes Y206F, D58A and Y318A are stable, mutant enzyme D58N loses about 20% of its inital activity | Thermobifida fusca |
60 | - |
15 h, wild-type enzyme is stable, mutant enzyme D58A loses about 20% of initial activity, mutant enzymes Y206F, D58N, D58N and Y318A lose about 10% of the inital activity | Thermobifida fusca |
65 | - |
15 h, wild-type enzyme loses about 10% of initial activity, mutant enzyme D58N and Y206F loses about 20% of maximal activity, mutant enzyme D58A loses about 90% of initial activity, mutant enzyme Y318A loses about 30% of initial activity | Thermobifida fusca |
70 | - |
wild-type enzyme loses about 50% of initial activity, mutant enzyme D58A loses about 20% of initial activity, mutant enymes D58A, Y206F and Y318A nearly completely lose activity | Thermobifida fusca |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.02 | - |
2,4-dinitrophenyl beta-D-cellobioside | 50°C, wild-type enzyme Cel9A-68 | Thermobifida fusca | |
0.07 | - |
2,4-dinitrophenyl beta-D-cellobioside | 50°C, mutant enzyme E424A | Thermobifida fusca | |
0.7 | - |
2,4-dinitrophenyl beta-D-cellobioside | 50°C, mutant enzyme E424G | Thermobifida fusca |