Literature summary for 3.2.1.25 extracted from

Tailford, L.E.; Money, V.A.; Smith, N.L.; Dumon, C.; Davies, G.J.; Gilbert, H.J.
Mannose foraging by Bacteroides thetaiotaomicron: structure and specificity of the beta-mannosidase, BtMan2A (2007), J. Biol. Chem., 282, 11291-11299.

Search for more literature for 3.2.1.25

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli strain BL21	Bacteroides thetaiotaomicron

Crystallization (Commentary)

Crystallization (Comment)	Organism
sitting drop vapour diffusion method with either 0.15-0.35 M NaBr and 6-12% PEG 3350, 0.1 M bis-Tris propane, pH 7, or 0.08-0.2 M KSCN, and 8-15% PEG 3350, 0.1 M bis-Tris propane, pH 7	Bacteroides thetaiotaomicron

Protein Variants

Protein Variants	Comment	Organism
D199A	low activity	Bacteroides thetaiotaomicron
E462A	low activity	Bacteroides thetaiotaomicron
E555A	low activity	Bacteroides thetaiotaomicron
N461A	low activity	Bacteroides thetaiotaomicron
Q646A	low activity	Bacteroides thetaiotaomicron
W198A	low activity	Bacteroides thetaiotaomicron
W200A	low activity	Bacteroides thetaiotaomicron
W395A	low activity	Bacteroides thetaiotaomicron
W645A	low activity	Bacteroides thetaiotaomicron
Y537A	low activity	Bacteroides thetaiotaomicron

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.0028	-	2,4-dinitrophenyl-beta-D-mannopyranoside	mutant enzyme E462A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.015	-	2,4-dinitrophenyl-beta-D-mannopyranoside	mutant enzyme N461A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.017	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme W395A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.079	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme W198A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.088	-	2,4-dinitrophenyl-beta-D-mannopyranoside	wild type enzyme, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.089	-	2,4-dinitrophenyl-beta-D-mannopyranoside	mutant enzyme E555A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.18	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme E555A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.19	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme E462A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.19	-	p-nitrophenyl-beta-D-mannopyranoside	wild type enzyme, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.31	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme Q646A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.31	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme Y537A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.43	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme N461A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
10	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme W200A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
10	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme W645A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
11	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme D199A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron

Organism

Organism	UniProt	Comment	Textmining
Bacteroides thetaiotaomicron	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
TALON immobilized metal ion affinity chromatography, Q12 column chromatography, and S200 gel filtration	Bacteroides thetaiotaomicron

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2,4-dinitrophenyl-beta-D-mannopyranoside + H2O	-	Bacteroides thetaiotaomicron	2,4-dinitrophenol + alpha-D-mannose	-	?
p-nitrophenyl beta-D-mannopyranoside + H2O	-	Bacteroides thetaiotaomicron	p-nitrophenol + beta-D-mannose	-	?

Subunits

Subunits	Comment	Organism
homodimer	x-ray crystallography	Bacteroides thetaiotaomicron

Synonyms

Synonyms	Comment	Organism
GH2 beta-mannosidase	-	Bacteroides thetaiotaomicron
Man2A	-	Bacteroides thetaiotaomicron

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.00183	-	2,4-dinitrophenyl-beta-D-mannopyranoside	mutant enzyme E555A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.008	-	2,4-dinitrophenyl-beta-D-mannopyranoside	mutant enzyme E462A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.0097	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme E555A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.045	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme D199A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.092	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme E462A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.25	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme N461A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.478	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme W645A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.85	-	2,4-dinitrophenyl-beta-D-mannopyranoside	mutant enzyme N461A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
0.886	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme W395A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
3.5	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme W198A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
6.08	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme W395A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
15.9	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme Y537A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
20.38	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme W198A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
26.85	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme Y537A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
34.43	-	p-nitrophenyl-beta-D-mannopyranoside	mutant enzyme Q646A, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
128.1	-	p-nitrophenyl-beta-D-mannopyranoside	wild type enzyme, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
157	-	p-nitrophenyl-beta-D-mannopyranoside	wild type enzyme, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron
214.4	-	2,4-dinitrophenyl-beta-D-mannopyranoside	wild type enzyme, at 37°C in 43.5 mM sodium phosphate, 10 mM citric acid buffer, pH 5.6, containing 1 mg/ml bovine serum albumin	Bacteroides thetaiotaomicron

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Release 2023.1 (January 2023)