Cloned (Comment) | Organism |
---|---|
constitutive recombinant expression of His-tagged enzyme in Escherichia coli strain MC1061 | Thermotoga maritima |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged enzyme in complex with a branched tetrasaccharide O-beta-D-xylopyranosyl-(1->4)-O-alpha-L-arabinofuranosyl-(1->3)-O-beta-D-xylopyranosyl-(1->4)-O-beta-D-xylopyranoside, sitting drop vapor-diffusion method, mixing 0.0015 ml of protein solution containing 14 mg/ml protein, 50 mM Tris-HCl, pH 7.5, 50 mM NaCl, and 5.0% 2-mercaptoethanol, with an equal volume of the reservoir solution, 16.1°C, X-ray diffraction structure determination and analysis | Thermotoga maritima |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-arabinan + H2O | Thermotoga maritima | - |
? | - |
? | |
L-arabinan + H2O | Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 | - |
? | - |
? | |
L-arabinoxylan + H2O | Thermotoga maritima | - |
? | - |
? | |
L-arabinoxylan + H2O | Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 | - |
? | - |
? | |
additional information | Thermotoga maritima | a highly thermostable exo-acting hemicellulase that exhibits a relatively higher activity towards arabinan and arabinoxylan, compared with other glycoside hydrolase 51 family enzymes | ? | - |
? | |
additional information | Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 | a highly thermostable exo-acting hemicellulase that exhibits a relatively higher activity towards arabinan and arabinoxylan, compared with other glycoside hydrolase 51 family enzymes | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermotoga maritima | Q9WYB7 | - |
- |
Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 | Q9WYB7 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain MC1061 by nickel affinity chromatography, dialysis, and gel filtration, to homogeneity | Thermotoga maritima |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
1,5-alpha-L-arabinofuranohexaose + 5 H2O = 6 alpha-L-arabinofuranose | the enzyme hydrolyze the glycosidic bond via the double-displacement mechanism in the active site of a funnel-shaped pocket, catalytic residues are the acid/base Glu172 and the enzymatic nucleophile Glu281 | Thermotoga maritima |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-arabinan + H2O | - |
Thermotoga maritima | ? | - |
? | |
L-arabinan + H2O | - |
Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 | ? | - |
? | |
L-arabinoxylan + H2O | - |
Thermotoga maritima | ? | - |
? | |
L-arabinoxylan + H2O | - |
Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 | ? | - |
? | |
additional information | a highly thermostable exo-acting hemicellulase that exhibits a relatively higher activity towards arabinan and arabinoxylan, compared with other glycoside hydrolase 51 family enzymes | Thermotoga maritima | ? | - |
? | |
additional information | a highly thermostable exo-acting hemicellulase that exhibits a relatively higher activity towards arabinan and arabinoxylan, compared with other glycoside hydrolase 51 family enzymes | Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme is organized by two domains: a catalytic N-terminal (beta/alpha)8-barrel domain and a C-terminal beta-strand sandwich domain, three-dimensional structure, interaction at the dimer interface, overview | Thermotoga maritima |
Synonyms | Comment | Organism |
---|---|---|
alpha-L-arabinofuranosidase | - |
Thermotoga maritima |
TmAFase | - |
Thermotoga maritima |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | - |
- |
Thermotoga maritima |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
4.5 | - |
- |
Thermotoga maritima |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the glycoside hydrolase 51 family, GH51 | Thermotoga maritima |
additional information | tight domain associations found in the enzyme, such as an interdomain disulfide bond (Cys306 and Cys476) in each monomer, an extended arm (amino acids 374-385) at the dimer interface, and total 12 salt bridges in the hexamer, may account for the thermostability of the enzyme. One of the xylan binding determinants (Trp96) is identified in the active site, and a region of amino acids (374-385) protrudes out forming an obvious wall at the substrate-binding groove to generate a cavity. The altered cavity shape with a strong negative electrostatic distribution is likely related to the unique substrate preference of TmAFase towards branched polymeric substrates | Thermotoga maritima |