Application | Comment | Organism |
---|---|---|
industry | alpha-L-arabinanases are essential glycosyl hydrolases participating in the complete hydrolysis of hemicellulose, a natural resource for various industrial processes, such as bioethanol or pharmaceuticals production | Fusarium graminearum |
synthesis | alpha-L-arabinanases are essential glycosyl hydrolases participating in the complete hydrolysis of hemicellulose, a natural resource for various industrial processes, such as bioethanol or pharmaceuticals production | Fusarium graminearum |
Cloned (Comment) | Organism |
---|---|
gene fg03054.1, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain TOP10 | Fusarium graminearum |
Crystallization (Comment) | Organism |
---|---|
purified recombinant free enzyme and in complex with alpha-1,5-L-arabinobiose, hanging drop vapor diffusion method, mixing of 0.001 ml of protein solution, containing 7.8 mg/ml protein, with an equal volume of reservoir solution containing 10% PEG 20000, 100 mM Tris, pH 8.5, and 100 mM MgCl2, 20°C. Selenomethionine crystals are obtained from a solution containing 25% PEG 550 MME, 100 mM HEPES, pH 7.5, and at 11.6 mg/ml protein. The complex of selenomethionine-substituted crystals with alpha-L-arabinobiose is obtained from 22% PEG 4000, 100 mM HEPES, pH 7.5, and 50 mM ammonium sulfate, for 2 h at room temperature mixed with 1 mM alpha-methyl-1,5-arabinotetraose, the precursor of alpha-1,5-L-arabinose. X-ray diffraction structure determination and analysis at 1.85 A and 2.05 A resolution, respectively | Fusarium graminearum |
Protein Variants | Comment | Organism |
---|---|---|
E170A | site-directed mutagenesis of a catalytic residue, inactive mutant | Fusarium graminearum |
E242A | site-directed mutagenesis of a catalytic residue, the mutant shows 7% of wild-type enzyme activity | Fusarium graminearum |
Y337A | site-directed mutagenesis of a catalytic residue, inactive mutant | Fusarium graminearum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics | Fusarium graminearum | |
2.03 | - |
alpha-1,5-L-arabinohexaose | pH 5.0, 40°C, wild-type enzyme | Fusarium graminearum | |
3.56 | - |
alpha-1,5-L-arabinotriose | pH 5.0, 40°C, wild-type enzyme | Fusarium graminearum | |
3.96 | - |
alpha-1,5-L-arabinopentaose | pH 5.0, 40°C, wild-type enzyme | Fusarium graminearum | |
9.26 | - |
alpha-1,5-L-arabinotetraose | pH 5.0, 40°C, wild-type enzyme | Fusarium graminearum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
alpha-L-arabinan + H2O | Fusarium graminearum | the enzyme is highly active and displays a strict substrate specificity for linear alpha-1,5-L-arabinan | alpha-1,5-L-arabinobiose + ? | the enzyme releases alpha-1,5-L-arabinobiose from the nonreducing end of the polysaccharide | ? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Fusarium graminearum | - |
gene fg03054.1 | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain TOP10 by metal affinity and anion exchange chromatography | Fusarium graminearum |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
1,5-alpha-L-arabinofuranohexaose + 5 H2O = 6 alpha-L-arabinofuranose | catalytic mechanism and substrate binding structure, overview | Fusarium graminearum |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
113 | - |
substrate sugar beet arabinan | Fusarium graminearum |
1065 | - |
substrate sugar beet debranched alpha-1,5-L-arabinan | Fusarium graminearum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
alpha-1,5-L-arabinohexaose + H2O | - |
Fusarium graminearum | alpha-1,5-L-arabinobiose + alpha-1,5-L-arabinotetraose | - |
? | |
alpha-1,5-L-arabinopentaose + H2O | - |
Fusarium graminearum | alpha-1,5-L-arabinobiose + alpha-1,5-L-arabinotriose | - |
? | |
alpha-1,5-L-arabinotetraose + H2O | - |
Fusarium graminearum | alpha-1,5-L-arabinobiose | - |
? | |
alpha-1,5-L-arabinotriose + H2O | - |
Fusarium graminearum | alpha-1,5-L-arabinobiose + alpha-L-arabinose | - |
? | |
alpha-L-arabinan + H2O | the enzyme is highly active and displays a strict substrate specificity for linear alpha-1,5-L-arabinan | Fusarium graminearum | alpha-1,5-L-arabinobiose + ? | the enzyme releases alpha-1,5-L-arabinobiose from the nonreducing end of the polysaccharide | ? | |
alpha-L-arabinan + H2O | - |
Fusarium graminearum | oligo-alpha-1,5-L-arabinose + ? | the enzyme releases alpha-1,5-L-arabinobiose from the nonreducing end of the polysaccharide | ? | |
additional information | no activity with 4-nitrophenyl-alpha-L-arabinofuranoside, wheat arabinoxylan, and alpha-L-arabinobiose, the configuration of bound arabinobiose is consistent with the retaining mechanism proposed for the GH93 family | Fusarium graminearum | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | Arb93A has a six-bladed beta-propeller fold | Fusarium graminearum |
Synonyms | Comment | Organism |
---|---|---|
Arb93A | - |
Fusarium graminearum |
exo-alpha-L-arabinanase | - |
Fusarium graminearum |
More | the enzyme belongs to the glycosylhydrolase family GH93 | Fusarium graminearum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
- |
Fusarium graminearum |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | 60 | over 80% of maximal activity at 30-50°C | Fusarium graminearum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
81000 | - |
alpha-1,5-L-arabinopentaose | pH 5.0, 40°C, wild-type enzyme | Fusarium graminearum | |
94000 | - |
alpha-1,5-L-arabinohexaose | pH 5.0, 40°C, wild-type enzyme | Fusarium graminearum | |
105000 | - |
alpha-1,5-L-arabinotetraose | pH 5.0, 40°C, wild-type enzyme | Fusarium graminearum | |
113000 | - |
alpha-1,5-L-arabinotriose | pH 5.0, 40°C, wild-type enzyme | Fusarium graminearum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
- |
Fusarium graminearum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
3 | 9 | over 80% of maximal activity at pH 4.0-6.0 | Fusarium graminearum |
General Information | Comment | Organism |
---|---|---|
physiological function | alpha-L-arabinanases are essential glycosyl hydrolases participating in the complete hydrolysis of hemicellulose | Fusarium graminearum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
11300 | - |
alpha-1,5-L-arabinotetraose | pH 5.0, 40°C, wild-type enzyme | Fusarium graminearum | |
20500 | - |
alpha-1,5-L-arabinopentaose | pH 5.0, 40°C, wild-type enzyme | Fusarium graminearum | |
31700 | - |
alpha-1,5-L-arabinotriose | pH 5.0, 40°C, wild-type enzyme | Fusarium graminearum | |
46600 | - |
alpha-1,5-L-arabinohexaose | pH 5.0, 40°C, wild-type enzyme | Fusarium graminearum |