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Literature summary for 3.2.1.23 extracted from

  • Campuzano, S.; Serra, B.; Llull, D.; Garcia, J.L.; Garcia, P.
    Cloning, expression, and characterization of a peculiar choline-binding beta-galactosidase from Streptococcus mitis (2009), Appl. Environ. Microbiol., 75, 5972-5980.
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
dithiothreitol 32% increase of activity by 10 mM dithiothreitol Streptococcus mitis

Cloned(Commentary)

Cloned (Comment) Organism
expressed as His-tagged enzyme in Escherichia coli MC4100 cells Streptococcus mitis

General Stability

General Stability Organism
the enzyme remains highly stable in solution or immobilized at room temperature in the absence of protein stabilizers, the His tag does not affect activity Streptococcus mitis

Inhibitors

Inhibitors Comment Organism Structure
Cu2+ 46% residual activity at 5 mM Streptococcus mitis
Fe2+ 68% residual activity at 5 mM Streptococcus mitis
additional information enzyme activity is not inhibited by D-glucose, even at 200 mM Streptococcus mitis
SDS 1% (w/v) SDS strongly inhibits the ONPGase activity Streptococcus mitis
Zn2+ 60% residual activity at 5 mM Streptococcus mitis

Localization

Localization Comment Organism GeneOntology No. Textmining
cell wall
-
Streptococcus mitis 5618
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
252000
-
analytical ultracentrifugation Streptococcus mitis
264000
-
2 * 264000, full-length enzyme, estimated from SDS-PAGE Streptococcus mitis

Organism

Organism UniProt Comment Textmining
Streptococcus mitis
-
-
-
Streptococcus mitis NCTC 12261T
-
-
-

Purification (Commentary)

Purification (Comment) Organism
the mature enzyme is purified by DEAE-cellulose column chromatography and Sephacryl S-300 gel filtration, the His-tagged enzyme is purified by Bio-Scale Mini Profinity immobilized metal affinity chromatography Streptococcus mitis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
898
-
mature enzyme from crude extract, at 30°C in 0.1 M sodium phosphate, pH 7.0, 10 mM KCl, 1 mM MgSO4, 5 mM mercaptoethanol Streptococcus mitis
1093
-
His-tagged enzyme from crude extract, at 30°C in 0.1 M sodium phosphate, pH 7.0, 10 mM KCl, 1 mM MgSO4, 5 mM mercaptoethanol Streptococcus mitis
2444
-
mature enzyme after 2.7fold purification, at 30°C in 0.1 M sodium phosphate, pH 7.0, 10 mM KCl, 1 mM MgSO4, 5 mM mercaptoethanol Streptococcus mitis
2969
-
His-tagged enzyme after 2.7fold purification, at 30°C in 0.1 M sodium phosphate, pH 7.0, 10 mM KCl, 1 mM MgSO4, 5 mM mercaptoethanol Streptococcus mitis

Storage Stability

Storage Stability Organism
-20°C, 20 mM phosphate buffer, pH 6.9 without the addition of protein stabilizers, at least 2 years, no loss of activity Streptococcus mitis
25°C, 20 mM phosphate buffer, pH 6.9 without the addition of protein stabilizers, 55 days, no loss of activity Streptococcus mitis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-nitrophenyl beta-D-galactopyranoside + H2O about 83% activity compared to 4-nitrophenyl-beta-D-galactopyranoside Streptococcus mitis 2-nitrophenol + beta-D-galactose
-
?
2-nitrophenyl beta-D-galactopyranoside + H2O about 83% activity compared to 4-nitrophenyl-beta-D-galactopyranoside Streptococcus mitis NCTC 12261T 2-nitrophenol + beta-D-galactose
-
?
4-nitrophenyl beta-D-galactopyranoside + H2O 100% activity Streptococcus mitis 4-nitrophenol + beta-D-galactose
-
?
4-nitrophenyl beta-D-galactopyranoside + H2O 100% activity Streptococcus mitis NCTC 12261T 4-nitrophenol + beta-D-galactose
-
?
lactose + H2O about 36% activity compared to 4-nitrophenyl-beta-D-galactopyranoside Streptococcus mitis D-galactose + D-glucose
-
?
lactose + H2O about 36% activity compared to 4-nitrophenyl-beta-D-galactopyranoside Streptococcus mitis NCTC 12261T D-galactose + D-glucose
-
?
lactulose + H2O about 0.7% activity compared to 4-nitrophenyl-beta-D-galactopyranoside Streptococcus mitis ?
-
?
lactulose + H2O about 0.7% activity compared to 4-nitrophenyl-beta-D-galactopyranoside Streptococcus mitis NCTC 12261T ?
-
?

Subunits

Subunits Comment Organism
monomer 2 * 264000, full-length enzyme, estimated from SDS-PAGE Streptococcus mitis

Synonyms

Synonyms Comment Organism
beta-D-galactoside galactohydrolase
-
Streptococcus mitis
ONPGase
-
Streptococcus mitis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30 40
-
Streptococcus mitis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
above 50°C the activity rapidly decreases Streptococcus mitis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6 6.5
-
Streptococcus mitis