Application | Comment | Organism |
---|---|---|
industry | DNA shuffling improves the catalytic activity and alkaline pH stability of Thermobifida fusca xylanase A toward the hydrolysis of xylan evolved enzyme 2TfxA98 can meet the requirements of biobleaching by its stability and activation under conditions of alkaline and high temperatures | Thermobifida fusca |
Cloned (Comment) | Organism |
---|---|
TfxA-cat gene cleaved by BamHI and PstI and ligated with vector pQE30 | Thermobifida fusca |
Protein Variants | Comment | Organism |
---|---|---|
G217L | increases activity and alkaline pH stability to some extent | Thermobifida fusca |
G25P | mainly leads to increase in alkaline pH stability without improving the specific activity | Thermobifida fusca |
I91T | results in the main contribution to catalytic activity rather than alkaline pH stability | Thermobifida fusca |
T21A | mainly leads to increase in alkaline pH stability without improving the specific activity | Thermobifida fusca |
T21A/G25P/V87P/J91T/G217L | mutant 2TfxA98, with approximately 12fold increased kcat/Km and 4.5fold decreased Km compared with its parent. Mutant 2TfxA98 inherits its thermostability from parent TfxA and enhances its alkaline pH stability through DNA shuffling. 2Tfx98 has significantly improved catalytic activities in performing the xylan hydrolysis. It is most active at 75°C, almost the same as the parental TfxA, but 2TfxA98 produces 1.9 times more reducing sugar than TfxA at 75°C of pH 9.0 | Thermobifida fusca |
V87P | significantly improves both the activity and alkaline pH stability | Thermobifida fusca |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermobifida fusca | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant TfxA and the mutants purified to homogeneity on nickel-chelating column | Thermobifida fusca |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
506 | - |
mutant G25P | Thermobifida fusca |
565 | - |
mutant T21A | Thermobifida fusca |
686 | - |
wild-type | Thermobifida fusca |
851 | - |
mutant G217L | Thermobifida fusca |
896 | - |
mutant I91T | Thermobifida fusca |
940 | - |
mutant V87P | Thermobifida fusca |
1108 | - |
mutant 2TfxA98 | Thermobifida fusca |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
xylan + H2O | - |
Thermobifida fusca | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
xylanase A | - |
Thermobifida fusca |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
both recombinant TfxA and mutant 2TfxA98 are stable at 70°C during 30 min incubation, although the mutant is slightly less stable than the parent at 70°C after 30 min of incubation | Thermobifida fusca |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
86 | - |
xylan | wild-type | Thermobifida fusca | |
225 | - |
xylan | mutant 2TfxA98 | Thermobifida fusca |