Literature summary for 3.2.1.99 extracted from

Alhassid, A.; Ben-David, A.; Tabachnikov, O.; Libster, D.; Naveh, E.; Zolotnitsky, G.; Shoham, Y.; Shoham, G.
Crystal structure of an inverting GH 43 1,5-alpha-L-arabinanase from Geobacillus stearothermophilus complexed with its substrate (2009), Biochem. J., 422, 73-82.

Cloned(Commentary)

Cloned (Comment)	Organism
gene abnB, expression of wild-type and mutant enzymes	Geobacillus stearothermophilus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant wild-type enzyme, and mutant enzyme E201A in complex with arabinotriose, hanging drop vapour diffusion, 0.0025-0.003 ml of protein solution with 6.5-13 mg/ml protein is mixed with an equal volume of precipitant solution containing 1.7 M lithium sulfate, 0.1 M Tris buffer, pH 7.5, and 15% v/v glycerol for the wild-type enzyme, and 1.9 M lithium sulfate, 0.1 M Tris buffer, pH 8.5, and 4% w/v PEG 400 for the mutant enzyme, X-ray diffraction structure determination and analysis, molecular replacement	Geobacillus stearothermophilus

Crystallization (Comment)

Organism

purified recombinant wild-type enzyme, and mutant enzyme E201A in complex with arabinotriose, hanging drop vapour diffusion, 0.0025-0.003 ml of protein solution with 6.5-13 mg/ml protein is mixed with an equal volume of precipitant solution containing 1.7 M lithium sulfate, 0.1 M Tris buffer, pH 7.5, and 15% v/v glycerol for the wild-type enzyme, and 1.9 M lithium sulfate, 0.1 M Tris buffer, pH 8.5, and 4% w/v PEG 400 for the mutant enzyme, X-ray diffraction structure determination and analysis, molecular replacement

Geobacillus stearothermophilus

Protein Variants

Protein Variants	Comment	Organism
D147A	site-directed mutagenesis, structure comparison with the wild-type enzyme	Geobacillus stearothermophilus
E201A	site-directed mutagenesis, structure comparison with the wild-type enzyme	Geobacillus stearothermophilus

Organism

Organism	UniProt	Comment	Textmining
Geobacillus stearothermophilus	B3EYM8	gene abnB	-
Geobacillus stearothermophilus T-6	B3EYM8	gene abnB	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant enzymes	Geobacillus stearothermophilus

Reaction

Reaction	Comment	Organism	Reaction ID
alpha-L-Ara-(1-5)-O-alpha-L-Ara-(1-5)-O-alpha-L-Ara-(1-5)-O-alpha-L-Ara-(1-5)-O-alpha-L-Ara + H2O = alpha-L-Ara-(1-5)-O-alpha-L-Ara-(1-5)-O-alpha-L-Ara + alpha-L-arabinofuranosyl-(1-5)-O-alpha-L-arabinofuranose	three catalytic carboxylates: Asp27, the general base, Glu201, the general acid, and Asp147, the pKa modulator, substrate binding structure and catalytic mechanism, overview	Geobacillus stearothermophilus	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
linear arabinan + H2O	-	Geobacillus stearothermophilus	?	-	?
linear arabinan + H2O	-	Geobacillus stearothermophilus T-6	?	-	?

Synonyms

Synonyms	Comment	Organism
More	the enzyme belongs to the glycoside hydrolase family 43, GH43	Geobacillus stearothermophilus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
40	-	assay at	Geobacillus stearothermophilus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	assay at	Geobacillus stearothermophilus