Literature summary for 3.2.1.145 extracted from

Godoy, A.S.; de Lima, M.Z.; Camilo, C.M.; Polikarpov, I.
Crystal structure of a putative exo-beta-1,3-galactanase from Bifidobacterium bifidum S17 (2016), Acta Crystallogr. Sect. F, 72, 288-293 .

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Cloned(Commentary)

Cloned (Comment)	Organism
gene BBNG_01788, sequence comparisons, recombinant expression of Trx-His-tagged enzyme in Escherichia coli strain Rosetta (DE3)	Bifidobacterium bifidum

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme, vapour diffusion method, mixing of 0.001 ml of 80 mg/ml protein in 0.02 M HEPES, pH 7.5, and 0.15 M NaCl, with 0.001 ml of reservoir solution containing 0.05 M calcium chloride dihydrate,0.1 M bis-Tris pH 6.5, 30% v/v polyethylene glycol monomethyl ether 550, and equilibration against 80.00 ml of reservoir solution, 17°C, X-ray diffraction structure determination and analysis at 1.4 A resolution, molecular replacement using using domain I of the Bacteroides thetaiotaomicron glycosyl hydrolase as search model, PDB ID 3nqh, modelling	Bifidobacterium bifidum

Crystallization (Comment)

Organism

purified recombinant enzyme, vapour diffusion method, mixing of 0.001 ml of 80 mg/ml protein in 0.02 M HEPES, pH 7.5, and 0.15 M NaCl, with 0.001 ml of reservoir solution containing 0.05 M calcium chloride dihydrate,0.1 M bis-Tris pH 6.5, 30% v/v polyethylene glycol monomethyl ether 550, and equilibration against 80.00 ml of reservoir solution, 17°C, X-ray diffraction structure determination and analysis at 1.4 A resolution, molecular replacement using using domain I of the Bacteroides thetaiotaomicron glycosyl hydrolase as search model, PDB ID 3nqh, modelling

Bifidobacterium bifidum

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
67300	-	recombinant detagged enzyme, gel filtration	Bifidobacterium bifidum

Organism

Organism	UniProt	Comment	Textmining
Bifidobacterium bifidum	E4VCC5	-	-
Bifidobacterium bifidum NCIMB 41171	E4VCC5	-	-
Bifidobacterium bifidum S17	E4VCC5	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant Trx-His-tagged enzyme from Escherichia coli strain Rosetta (DE3) by nickel affinity chromatography, tag cleavage through TEV protease, and another step of nickel affinity chromatography, followed by gel filtration, to homogeneity	Bifidobacterium bifidum

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
additional information	the recombinant BbGal43A shows activity towards synthetic galactosides	Bifidobacterium bifidum	?	-	?
additional information	the recombinant BbGal43A shows activity towards synthetic galactosides	Bifidobacterium bifidum S17	?	-	?
additional information	the recombinant BbGal43A shows activity towards synthetic galactosides	Bifidobacterium bifidum NCIMB 41171	?	-	?

Subunits

Subunits	Comment	Organism
dimer	2 * 34000, recombinant detagged enzyme, SDS-PAGE	Bifidobacterium bifidum
More	crystal contact analysis predicts the formation of a dimer in solution. The structure shows a single-domain molecule, differing from known homologues. BbGal43A has a typical fivefold beta-propeller fold, with the propellers formed by four beta-sheets	Bifidobacterium bifidum

Synonyms

Synonyms	Comment	Organism
BbGal43A	-	Bifidobacterium bifidum
exo-beta-1,3-galactanase	-	Bifidobacterium bifidum

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the glycosyl hydrolase family 43, GH43. Structure comparisons reveal that structural and sequence homology between BbGal43A, Clostridium thermocellum Ct1,3Gal43A (PDB ID 3vsf), and Bacteroides thetaiotaomicron BT2959 (PDB ID 3nqh) indicate that all three enzymes might be different types of exo-beta-1,3-galactanase	Bifidobacterium bifidum