Literature summary for 3.2.1.86 extracted from

Stepper, J.; Dabin, J.; Eklof, J.M.; Thongpoo, P.; Kongsaeree, P.; Taylor, E.J.; Turkenburg, J.P.; Brumer, H.; Davies, G.J.
Structure and activity of the Streptococcus pyogenes family GH1 6-phospho-beta-glucosidase SPy1599 (2013), Acta Crystallogr. Sect. D, 69, 16-23.

Cloned(Commentary)

Cloned (Comment)	Organism
gene Spy1599, sequence comparisons, expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3)	Streptococcus pyogenes

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystals with C2, P2 twinned, and P2 not twinned structures, hanging-drop vapour diffusion method, mixing equal volumes of 20 mg/ml protein with 0.1 M Bis-Tris propane, pH 7.5, 0.2 M sodium fluoride or sodium bromide, 20% PEG 3350 at 19°C, X-ray diffraction structure determination and analysis at 2.51-2.61 A resolution, three-dimensional structure analysis of the enzyme in a number of crystal forms complicated by unusual crystallographic twinning, the enzyme shows a classical (beta/alpha)8-barrel structure, molecular replacement method	Streptococcus pyogenes

Crystallization (Comment)

Organism

crystals with C2, P2 twinned, and P2 not twinned structures, hanging-drop vapour diffusion method, mixing equal volumes of 20 mg/ml protein with 0.1 M Bis-Tris propane, pH 7.5, 0.2 M sodium fluoride or sodium bromide, 20% PEG 3350 at 19°C, X-ray diffraction structure determination and analysis at 2.51-2.61 A resolution, three-dimensional structure analysis of the enzyme in a number of crystal forms complicated by unusual crystallographic twinning, the enzyme shows a classical (beta/alpha)8-barrel structure, molecular replacement method

Streptococcus pyogenes

General Stability

General Stability	Organism
SPy1599 is subject to degradation during both storage and crystallization experiments	Streptococcus pyogenes

Inhibitors

Inhibitors	Comment	Organism	Structure
additional information	no inhibition by cyclohexylammonium or cyclohexylamine salt	Streptococcus pyogenes

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	Michaelis-Menten kinetics	Streptococcus pyogenes
0.73	-	4-nitrophenyl-beta-D-glucopyranoside 6-phosphate	pH 6.0, 37°C	Streptococcus pyogenes

Organism

Organism	UniProt	Comment	Textmining
Streptococcus pyogenes	Q1JK37	gene Spy1599	-
Streptococcus pyogenes M1 GAS SF370	Q1JK37	gene Spy1599	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, and gel filtration	Streptococcus pyogenes

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
4-nitrophenyl beta-D-glucopyranoside 6-phosphate + H2O	-	Streptococcus pyogenes	4-nitrophenol + beta-D-glucose 6-phosphate	-	?
4-nitrophenyl beta-D-glucopyranoside 6-phosphate + H2O	-	Streptococcus pyogenes M1 GAS SF370	4-nitrophenol + beta-D-glucose 6-phosphate	-	?
additional information	the enzyme SPy1599 contains a phosphate-binding pocket in the active site and has specific 6-phospho-beta-glucosidase activity. No or very poor activity with 4-nitrophenyl derivatives of galactose, glucose, xylose, galactose 6-phosphate, N-acetyl-D-galactosamine, and N-acetyl-D-glusosamine	Streptococcus pyogenes	?	-	?
additional information	the enzyme SPy1599 contains a phosphate-binding pocket in the active site and has specific 6-phospho-beta-glucosidase activity. No or very poor activity with 4-nitrophenyl derivatives of galactose, glucose, xylose, galactose 6-phosphate, N-acetyl-D-galactosamine, and N-acetyl-D-glusosamine	Streptococcus pyogenes M1 GAS SF370	?	-	?

Subunits

Subunits	Comment	Organism
More	the enzyme shows a classical (beta/alpha)8-barrel structure	Streptococcus pyogenes

Synonyms

Synonyms	Comment	Organism
SPy1599	-	Streptococcus pyogenes

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Streptococcus pyogenes

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.243	-	4-nitrophenyl-beta-D-glucopyranoside 6-phosphate	pH 6.0, 37°C	Streptococcus pyogenes

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6	-	-	Streptococcus pyogenes

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the glycosyl hydrolase family 1, GH1	Streptococcus pyogenes
additional information	structure comparisons, active center and substrate specificity, overview	Streptococcus pyogenes
physiological function	the enzyme SPy1599 is involved in the intracellular degradation of 6-phosphoglycosides, which are likely to originate from import through one of the organism's many phosphoenolpyruvate phosphotransfer systems	Streptococcus pyogenes

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.333	-	4-nitrophenyl-beta-D-glucopyranoside 6-phosphate	pH 6.0, 37°C	Streptococcus pyogenes