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Literature summary for 3.2.1.23 extracted from

  • Di Lauro, B.; Strazzulli, A.; Perugino, G.; La Cara, F.; Bedini, E.; Corsaro, M.M.; Rossi, M.; Moracci, M.
    Isolation and characterization of a new family 42 beta-galactosidase from the thermoacidophilic bacterium Alicyclobacillus acidocaldarius: identification of the active site residues (2008), Biochim. Biophys. Acta, 1784, 292-301.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene lacB, construction of a genomic library, DNA and amino acid sequence determination and anaylsis, expression as GST-tagged enzyme in Escherichia coli strain BL21RB791 Alicyclobacillus acidocaldarius

Protein Variants

Protein Variants Comment Organism
E157G site-directed mutagenesis, altered kinetics compared to the wild-type enzyme Alicyclobacillus acidocaldarius
E313G site-directed mutagenesis, altered kinetics compared to the wild-type enzyme Alicyclobacillus acidocaldarius

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.11
-
2-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, mutant E157G Alicyclobacillus acidocaldarius
4.8
-
2-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, mutant E313G Alicyclobacillus acidocaldarius
4.9
-
4-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
5.2
-
4-nitrophenyl-alpha-L-arabinopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
5.5
-
2-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
7.3
-
2-nitrophenyl-beta-D-fucopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
18.2
-
lactulose pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
60.8
-
lactose pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
Alicyclobacillus acidocaldarius 5829
-

Metals/Ions

Metals/Ions Comment Organism Structure
additional information the enzyme is not affected by 5 mM of Mg2+, Mn2+, or Ca2+ Alicyclobacillus acidocaldarius

Organism

Organism UniProt Comment Textmining
Alicyclobacillus acidocaldarius Q0R5R5 subsp. rittmanii, a thermoacidophilic bacterium, strain ATCC 27009
-

Purification (Commentary)

Purification (Comment) Organism
native enzyme 46fold by anion exchange and hydrophobic interaction chromatography and gel filtration, recombinant GST-tagged enzyme from Escherichia coli strain BL21RB791 by heat treatment at 60°C and hydrophobic interaction chromatography to over 95% purity Alicyclobacillus acidocaldarius

Reaction

Reaction Comment Organism Reaction ID
beta-D-galactopyranosyl-(1-4)-beta-D-galactopyranosyl-(1-6)-beta-D-galactopyranosyl-(1-3)-beta-D-galactopyranose + 3 H2O = 4 beta-D-galactopyranose catalytic mechanism, active site structure,Glu157 acts as the acid/base in the catalytic reaction Alicyclobacillus acidocaldarius

Source Tissue

Source Tissue Comment Organism Textmining
additional information optimal growth pH is pH 4.0 Alicyclobacillus acidocaldarius
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Alicyclobacillus acidocaldarius
592
-
purified native enzyme Alicyclobacillus acidocaldarius

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-nitrophenyl beta-D-galactopyranoside + H2O
-
Alicyclobacillus acidocaldarius 2-nitrophenol + beta-D-galactose
-
?
2-nitrophenyl-beta-D-fucopyranoside + H2O
-
Alicyclobacillus acidocaldarius 2-nitrophenol + beta-D-fucose
-
?
4-nitrophenyl beta-D-galactopyranoside + H2O
-
Alicyclobacillus acidocaldarius 4-nitrophenol + beta-D-galactose
-
?
4-nitrophenyl-alpha-L-arabinopyranoside + H2O
-
Alicyclobacillus acidocaldarius 4-nitrophenol + alpha-L-arabinose
-
?
lactose + H2O
-
Alicyclobacillus acidocaldarius D-glucose + D-galactose
-
?
lactulose + H2O
-
Alicyclobacillus acidocaldarius D-fructose + D-galactose
-
?
additional information the enzyme is very specific for glycosides with an axial C4-OH at their non-reducing end, no activity with cellobiose, glucosides, or xylosides Alicyclobacillus acidocaldarius ?
-
?

Synonyms

Synonyms Comment Organism
Aabeta-gal
-
Alicyclobacillus acidocaldarius
More the enzyme belongs to the glycosyl hydrolase family 42 Alicyclobacillus acidocaldarius

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
65 70 assay at Alicyclobacillus acidocaldarius

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
35 85
-
Alicyclobacillus acidocaldarius

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
65
-
3 h, 90% remianing activity Alicyclobacillus acidocaldarius
80
-
half-life 6 min Alicyclobacillus acidocaldarius

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.21
-
2-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, mutant E313G Alicyclobacillus acidocaldarius
0.32
-
2-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, mutant E157G Alicyclobacillus acidocaldarius
212
-
lactose pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
380
-
lactulose pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
1345
-
2-nitrophenyl-beta-D-fucopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
1740
-
4-nitrophenyl-alpha-L-arabinopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
2657
-
2-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
2988
-
4-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5 6.5 assay at Alicyclobacillus acidocaldarius