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Literature summary 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)

EC Number Cloned (Comment) Organism
3.2.1.23 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

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.23 0.11
-
 2-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, mutant E157G Alicyclobacillus acidocaldarius
3.2.1.23 4.8
-
 2-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, mutant E313G Alicyclobacillus acidocaldarius
3.2.1.23 4.9
-
 4-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
3.2.1.23 5.2
-
 4-nitrophenyl-alpha-L-arabinopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
3.2.1.23 5.5
-
 2-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
3.2.1.23 7.3
-
 2-nitrophenyl-beta-D-fucopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
3.2.1.23 18.2
-
 lactulose pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
3.2.1.23 60.8
-
 lactose pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius

Localization

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

Metals/Ions

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

Organism

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

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.23 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

EC Number Reaction Comment Organism Reaction ID
3.2.1.23 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
a

Source Tissue

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

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.23 additional information
-
-
 Alicyclobacillus acidocaldarius
3.2.1.23 592
-
 purified native enzyme Alicyclobacillus acidocaldarius

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.23 2-nitrophenyl beta-D-galactopyranoside + H2O
-
 Alicyclobacillus acidocaldarius 2-nitrophenol + beta-D-galactose
-
 ?
3.2.1.23 2-nitrophenyl-beta-D-fucopyranoside + H2O
-
 Alicyclobacillus acidocaldarius 2-nitrophenol + beta-D-fucose
-
 ?
3.2.1.23 4-nitrophenyl beta-D-galactopyranoside + H2O
-
 Alicyclobacillus acidocaldarius 4-nitrophenol + beta-D-galactose
-
 ?
3.2.1.23 4-nitrophenyl-alpha-L-arabinopyranoside + H2O
-
 Alicyclobacillus acidocaldarius 4-nitrophenol + alpha-L-arabinose
-
 ?
3.2.1.23 lactose + H2O
-
 Alicyclobacillus acidocaldarius D-glucose + D-galactose
-
 ?
3.2.1.23 lactulose + H2O
-
 Alicyclobacillus acidocaldarius D-fructose + D-galactose
-
 ?
3.2.1.23 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

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.23 65 70 assay at Alicyclobacillus acidocaldarius

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.23 35 85
-
 Alicyclobacillus acidocaldarius

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.23 65
-
 3 h, 90% remianing activity Alicyclobacillus acidocaldarius
3.2.1.23 80
-
 half-life 6 min Alicyclobacillus acidocaldarius

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.23 0.21
-
 2-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, mutant E313G Alicyclobacillus acidocaldarius
3.2.1.23 0.32
-
 2-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, mutant E157G Alicyclobacillus acidocaldarius
3.2.1.23 212
-
 lactose pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
3.2.1.23 380
-
 lactulose pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
3.2.1.23 1345
-
 2-nitrophenyl-beta-D-fucopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
3.2.1.23 1740
-
 4-nitrophenyl-alpha-L-arabinopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
3.2.1.23 2657
-
 2-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius
3.2.1.23 2988
-
 4-nitrophenyl-beta-D-galactopyranoside pH 6.5, 65°C, wild-type enzyme Alicyclobacillus acidocaldarius

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.23 5.5 6.5 assay at Alicyclobacillus acidocaldarius