Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and mutant enzymes in Streptomyces lividans strain 1326 and secretion to the culture medium | Streptomyces coelicolor |
Crystallization (Comment) | Organism |
---|---|
purified recomnbinant enzyme free or complexed with mercury derivative, L-arabinose, or xylooligosaccharides, sitting drop vapour diffusion method, mixing of 0.0012 ml of 8.3 mg/ml protein in 2 mM Tris-HCl buffer, pH 7.0, and 20 mM NaCl, with 0.001 ml of reservoir solution containing 20% w/v PEG 3350 and 0.2 M tripotassium citrate, pH 8.3, 20°C, X-ray diffraction structure determination and analysis at 1.4-2.1 A resolution, model building and molecular refinement | Streptomyces coelicolor |
Protein Variants | Comment | Organism |
---|---|---|
D165E | site-directed mutagenesis | Streptomyces coelicolor |
D165N | site-directed mutagenesis | Streptomyces coelicolor |
D202N | site-directed mutagenesis, inactive mutant | Streptomyces coelicolor |
E324Q | site-directed mutagenesis | Streptomyces coelicolor |
E361Q | site-directed mutagenesis, inactive mutant | Streptomyces coelicolor |
N425Q | site-directed mutagenesis | Streptomyces coelicolor |
N462Q | site-directed mutagenesis, the mutant shows 30% reduced activity compared to the wild-type enzyme | Streptomyces coelicolor |
W233A | site-directed mutagenesis | Streptomyces coelicolor |
W233F | site-directed mutagenesis | Streptomyces coelicolor |
W233Y | site-directed mutagenesis | Streptomyces coelicolor |
W270A | site-directed mutagenesis, the mutant shows 70% reduced activity compared to the wild-type enzyme | Streptomyces coelicolor |
W270F | site-directed mutagenesis, the mutant shows 70% reduced activity compared to the wild-type enzyme | Streptomyces coelicolor |
W270Y | site-directed mutagenesis, the mutant shows 50% reduced activity compared to the wild-type enzyme | Streptomyces coelicolor |
Y424A | site-directed mutagenesis | Streptomyces coelicolor |
Y424F | site-directed mutagenesis | Streptomyces coelicolor |
Y424W | site-directed mutagenesis | Streptomyces coelicolor |
Y461A | site-directed mutagenesis, the mutant shows 20% reduced activity compared to the wild-type enzyme | Streptomyces coelicolor |
Y461F | site-directed mutagenesis, the mutant shows 80% reduced activity compared to the wild-type enzyme | Streptomyces coelicolor |
Y461W | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Streptomyces coelicolor |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of mutant enzymes, overview | Streptomyces coelicolor | |
1.9 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | ph 5.5, 37°C, recombinant wild-type enzyme | Streptomyces coelicolor | |
7.3 | - |
wheat L-arabinoxylan | ph 5.5, 37°C, recombinant wild-type enzyme | Streptomyces coelicolor |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
extracellular | - |
Streptomyces coelicolor | - |
- |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-arabinoxylan + H2O | Streptomyces coelicolor | the enzyme efficiently removes the alpha-L-arabinosyl substituents from arabinoxylan | ? | - |
? | |
L-arabinoxylan + H2O | Streptomyces coelicolor A3(2) | the enzyme efficiently removes the alpha-L-arabinosyl substituents from arabinoxylan | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces coelicolor | O54161 | - |
- |
Streptomyces coelicolor A3(2) | O54161 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Streptomyces lividans strain 1326 culture medium by affinity chromatography using lactosyl-Sepharose, dialysis and ultrafiltration | Streptomyces coelicolor |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl alpha-L-arabinofuranoside + H2O | - |
Streptomyces coelicolor | 4-nitrophenol + L-arabinofuranose | - |
? | |
4-nitrophenyl alpha-L-arabinofuranoside + H2O | - |
Streptomyces coelicolor A3(2) | 4-nitrophenol + L-arabinofuranose | - |
? | |
L-arabinoxylan + H2O | the enzyme efficiently removes the alpha-L-arabinosyl substituents from arabinoxylan | Streptomyces coelicolor | ? | - |
? | |
L-arabinoxylan + H2O | the enzyme efficiently removes the alpha-L-arabinosyl substituents from arabinoxylan | Streptomyces coelicolor A3(2) | ? | - |
? | |
additional information | substrate specificity and substrate-binding mechanism, overview | Streptomyces coelicolor | ? | - |
? | |
additional information | substrate specificity and substrate-binding mechanism, overview | Streptomyces coelicolor A3(2) | ? | - |
? | |
wheat L-arabinoxylan + H2O | the enzyme efficiently removes the alpha-L-arabinosyl substituents from arabinoxylan | Streptomyces coelicolor | ? | - |
? | |
wheat L-arabinoxylan + H2O | the enzyme efficiently removes the alpha-L-arabinosyl substituents from arabinoxylan | Streptomyces coelicolor A3(2) | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
exo-alpha-L-arabinofuranosidase | UniProt | Streptomyces coelicolor |
ScAraf62A | - |
Streptomyces coelicolor |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 45 | assay at | Streptomyces coelicolor |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.02 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | ph 5.5, 37°C, recombinant wild-type enzyme | Streptomyces coelicolor | |
0.3 | - |
wheat L-arabinoxylan | ph 5.5, 37°C, recombinant wild-type enzyme | Streptomyces coelicolor |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.5 | 6 | assay at | Streptomyces coelicolor |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the family 62 glycoside hydrolase, GH62. The crystal structure comprises a carbohydrate-binding module family 13 domain at its N-terminus and a catalytic domain at its C-terminus | Streptomyces coelicolor |
additional information | the catalytic domain is a five-bladed beta-propeller consisting of five radially oriented anti-parallel beta-sheets, substrate binding at five subsites in the catalytic cleft and an L-arabinose-binding pocket at the bottom of the cleft. Asp202 and Glu361 are catalytic residues, and Trp270, Tyr461, and Asn462 are involved in the substrate-binding site for discriminating the substrate structures | Streptomyces coelicolor |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.011 | - |
4-nitrophenyl-alpha-L-arabinofuranoside | pH 5.5, 37°C, recombinant wild-type enzyme | Streptomyces coelicolor | |
0.042 | - |
wheat L-arabinoxylan | pH 5.5, 37°C, recombinant wild-type enzyme | Streptomyces coelicolor |