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Information on EC 3.2.1.23 - beta-galactosidase and Organism(s) Escherichia coli and UniProt Accession P00722
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3.2.1.23 beta-galactosidaseIUBMB Comments
Some enzymes in this group hydrolyse alpha-L-arabinosides; some animal enzymes also hydrolyse beta-D-fucosides and beta-D-glucosides; cf. EC 3.2.1.108 lactase.
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Word Map
- 3.2.1.23
- lacz
- adenovirus
- lysosomal
- lactose
- endothelial
- oligosaccharide
- artery
- phage
-
adenovirus-mediated
- beta-glucosidase
-
immunoassay
-
retroviral
- beta-glucuronidase
-
herpes
- lambda
- simplex
- luciferase
- cytomegalovirus
- neuraminidase
- viruses
- hydrolases
- bacteriophage
- glycosidases
- gm1
- liposome
- ganglioside
- p16ink4a
- alpha-galactosidase
- alpha-mannosidase
-
osteogenic
- vaccinia
-
adeno-associated
-
replication-defective
- n-acetyl-beta-glucosaminidase
- runx2
- sialidase
- alpha-glucosidase
-
sialic
-
galactosylation
-
plaque-forming
-
osteoblast
- kluyveromyces
- telomerase
- analysis
- degradation
- diagnostics
-
lysogens
- beta-hexosaminidase
- keratan
-
vector-mediated
- biotechnology
- molecular biology
- synthesis
- environmental protection
- ganciclovir
- nutrition
-
transglycosylation
- medicine
- industry
-
nonviral
- food industry
- biofuel production
The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
Synonyms
beta-galactosidase, beta-gal, beta-d-galactosidase, senescence-associated beta-galactosidase, endo-beta-galactosidase, bglap, sa-beta-gal, acid beta-galactosidase, beta galactosidase, betagal, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
Acid beta-galactosidase
-
-
-
-
beta-D-galactoside galactohydrolase
-
-
-
-
beta-D-glactanase
-
-
-
-
beta-D-lactosidase
-
-
-
-
beta-galase
-
-
-
-
beta-lactosidase
-
-
-
-
driselase
-
-
-
-
Exo-(1-->4)-beta-D-galactanase
-
-
-
-
exo-beta-(1->3)-D-galactanase
-
-
-
-
galactosidase, beta
-
-
-
-
Hydrolact
-
-
-
-
lactase
-
-
-
-
lactosylceramidase II
-
-
-
-
Lactozym
-
-
-
-
Maxilact
-
-
-
-
Oryzatym
-
-
-
-
p-nitrophenyl beta-galactosidase
-
-
-
-
S 2107
-
-
-
-
SR12 protein
-
-
-
-
Sumiklat
-
-
-
-
Trilactase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hydrolysis of terminal non-reducing beta-D-galactose residues in beta-D-galactosides
mechanism
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of O-glycosyl bond
-
-
-
-
PATHWAY SOURCE
PATHWAYS
BRENDA
KEGG
-
-
SYSTEMATIC NAME
IUBMB Comments
beta-D-galactoside galactohydrolase
Some enzymes in this group hydrolyse alpha-L-arabinosides; some animal enzymes also hydrolyse beta-D-fucosides and beta-D-glucosides; cf. EC 3.2.1.108 lactase.
CAS REGISTRY NUMBER
COMMENTARY
9031-11-2
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2-nitrophenyl beta-D-galactopyranoside + H2O
2-nitrophenol + beta-D-galactose
-
-
-
?
2-nitrophenyl beta-D-galactopyranoside + H2O
2-nitrophenol + D-galactose
-
-
-
?
4-nitrophenyl beta-D-galactopyranoside + H2O
4-nitrophenol + beta-D-galactose
-
-
-
?
4-nitrophenyl beta-D-galactopyranoside + H2O
4-nitrophenol + D-galactose
-
-
-
?
(2R)-glycerol-O-D-beta-galactopyranoside + H2O
beta-D-galactose + glycerol
-
-
-
-
?
2,4-dinitrophenyl-beta-D-galactoside + H2O
2,4-dinitrophenol + beta-D-galactose
-
-
-
-
?
2,4-dinitrophenyl-beta-D-glucoside + H2O
2,4-dinitrophenol + beta-D-glucose
-
activity is 10000 times lower than with 2,4-dinitrophenyl-beta-D-galactoside
-
-
?
2-nitrophenyl beta-D-galactopyranoside + H2O
2-nitrophenol + beta-D-galactose
-
-
-
-
?
2-phenylethyl-beta-D-galactoside + H2O
2-phenylethanol + beta-D-galactose
-
-
-
-
?
4-nitrophenyl beta-D-galactoside + H2O
4-nitrophenol + beta-D-galactose
-
the enzyme catalyses both hydrolysis and transglycosylation
-
-
?
9H-(1,3-dichloro-9,9-dimethylacridin-2-one-7-yl) beta-D-galactopyranoside + H2O
9H-(1,3-dichloro-9,9-dimethylacridin-2-one-7-ol) + beta-D-galactose
-
-
-
-
?
9H-(1,3-dichloro-9,9-dimethylacridin-2-one-7-yl)-beta-D-galactoside + H2O
?
-
assays are performed on individual Escherichia coli beta-galactosidase molecules at 2 different concentrations of the substrate DDAO-beta-D-galactoside using a free zone capillary electrophoresis-based protocol with an in-laboratory-constructed instrument utilizing laser-induced fluorescence detection
-
-
?
chlorophenol red beta-D-galactopyranoside + H2O
chlorophenol red + beta-D-galactose
-
-
-
-
?
fluorescein-beta-D-digalactopyranoside + H2O
fluorescein + beta-D-galactose
-
-
-
-
?
methyl salicylate beta-D-galactoside + H2O
methyl salicylate + beta-D-galactose
-
-
-
-
?
p-nitrophenyl alpha-L-arabinoside + H2O
p-nitrophenol + alpha-L-arabinose
-
-
-
-
?
p-nitrophenyl-beta-D-galactoside + H2O
p-nitrophenol + beta-D-galactose
-
-
-
-
?
thio-(o-nitrophenyl)-beta-D-galactoside + H2O
thio-(o-nitrophenol) + beta-D-galactose
-
-
-
-
?
o-nitrophenyl-beta-D-galactoside + H2O
o-nitrophenol + beta-D-galactose
-
-
-
-
?
o-nitrophenyl-beta-D-galactoside + H2O
o-nitrophenol + beta-D-galactose
-
no hydrolysis activity is observed in 1,4-dioxane, methylsulphoxide, N,N-dimethylformamide, or pyridine
-
-
?
additional information
?
-
-
transgalactosylation with o-nitrophenyl-beta-D-galactoside or phenyl-beta-D-galactoside as galactosyl donor and methanol, glycerol, 2-mercaptoethanol, Tris or 2-phenylethyl alcohol as acceptor
-
-
?
additional information
?
-
-
synthesis of beta-D-galactosides by the direct condensation of free galactose and alcohols
-
-
?
additional information
?
-
-
the enzyme also exhibits transglycosylation activity with lactose as acceptor
-
-
?
additional information
?
-
-
the selectivity hexanol vs. water increases with increasing water activity in the reaction medium. It seems that the ideal condition for the transglycosylation reaction is a water activity close to 1.0
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
Ca2+
Co2+
-
0.1 mM, activation in a system containing Na+ at optimal concentration
Fe2+
K+
Mg2+
Mn2+
-
0.01-0.1 mM, activation in a system containing Na+ at optimal concentration
Na+
-
activates hydrolysis of o-nitrophenyl-beta-D-galactoside more effectively than K+
Ni2+
-
0.1 mM, activation in a system containing Na+ at optimal concentration
Zn2+
-
0.01-0.1 mM, activation in a system containing Na+ at optimal concentration
Mg2+
beta-galactosidase requires Mg2+ for full catalytic activity, one Mg2+ is at each active site but 3-4 others bind to other parts of each subunit
Mg2+
the active site of beta-galactosidase contains a Mg2+ ion ligated by Glu-416, His-418 and Glu-461 plus three water molecules
Ca2+
-
0.01 mM, activation at low concentrations in a system containing Na+ at optimal concentration
Fe2+
-
0.01-0.1 mM, activation in a system containing Na+ at optimal concentration
K+
-
activates hydrolysis of p-nitrophenyl-beta-D-galactoside and p-nitrophenyl alpha-L-arabinoside more effectively than Na+
Mg2+
-
0.1-10 mM, activation in a system containing Na+ at optimal concentration
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
benzyl 3-deoxy-4-S-(beta-D-galactopyranosyl)-4-thio-beta-D-erythro-pentopyranoside
-
non-competitive inhibitor
benzyl 3-deoxy-4-S-(beta-D-galactopyranosyl)-4-thio-beta-D-threo-pentopyranoside
-
mixed-type inhibitor
NaCl
-
the activity of both soluble beta-Gal and encapsulated beta-Gal decrease with salt addition. While in the case of soluble beta-Gal at 150 mM NaCl the reaction rate has decreased to half of the control value, in the case of encapsulated bet-Gal the same inhibition level is reached at 300 mM NaCl
additional information
-
not inhibited by 2-propyl 3-deoxy-4-S-(beta-D-xylopyranosyl)-4-thio-alpha-D-xylohexopyranoside and 2-propyl 3-deoxy-4-S-(beta-D-xylopyranosyl)-4-thio-alpha-D-lyxohexopyranoside
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.04
2-nitrophenyl beta-D-galactopyranoside
wild type enzyme, in 30 mM TES buffer, 145 mM NaCl, pH 7.0 at 25°C
0.06
2-nitrophenyl beta-D-galactopyranoside
mutant enzyme H418N, in 30 mM TES buffer, 0.01 mM Mg2+, 150 mM Na+, pH 7.0 at 25°C
30.6
2-nitrophenyl beta-D-galactopyranoside
mutant enzyme H418F, in 30 mM TES buffer, 10 mM Mg2+, 150 mM Na+, pH 7.0 at 25°C
104
2-nitrophenyl beta-D-galactopyranoside
mutant enzyme H418E, in 30 mM TES buffer, 0.01 mM Mg2+, 150 mM Na+, pH 7.0 at 25°C
0.04
4-nitrophenyl beta-D-galactopyranoside
mutant enzyme H418N, in 30 mM TES buffer, 0.01 mM Mg2+, 150 mM Na+, pH 7.0 at 25°C
0.12
4-nitrophenyl beta-D-galactopyranoside
wild type enzyme, in 30 mM TES buffer, 145 mM NaCl, pH 7.0 at 25°C
13.2
4-nitrophenyl beta-D-galactopyranoside
mutant enzyme H418E, in 30 mM TES buffer, 0.01 mM Mg2+, 150 mM Na+, pH 7.0 at 25°C
20.8
4-nitrophenyl beta-D-galactopyranoside
mutant enzyme H418F, in 30 mM TES buffer, 10 mM Mg2+, 150 mM Na+, pH 7.0 at 25°C
0.0836
2-nitrophenyl beta-D-galactopyranoside
-
enzyme encapsulated in a silica matrix
additional information
additional information
discussion of precautions and recommendations for measuring values of the rate constants and the associated enthalpies and entropies of enzymes with two catalytic steps by determining the effects of temperature on the kcat values
-
additional information
additional information
-
Km-values of chimeric enzymes
-
additional information
additional information
-
influence of temperature on Km-value of o-nitrophenyl-beta-D-galactoside
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
turnover numbers of chimeric galactosidases
-
4.9
2-nitrophenyl beta-D-galactopyranoside
mutant enzyme H418N, in 30 mM TES buffer, 0.01 mM Mg2+, 150 mM Na+, pH 7.0 at 25°C
5.4
2-nitrophenyl beta-D-galactopyranoside
mutant enzyme H418E, in 30 mM TES buffer, 0.01 mM Mg2+, 150 mM Na+, pH 7.0 at 25°C
10.4
2-nitrophenyl beta-D-galactopyranoside
mutant enzyme H418F, in 30 mM TES buffer, 10 mM Mg2+, 150 mM Na+, pH 7.0 at 25°C
90
2-nitrophenyl beta-D-galactopyranoside
wild type enzyme, in 30 mM TES buffer, 145 mM NaCl, pH 7.0 at 25°C
5.4
4-nitrophenyl beta-D-galactopyranoside
mutant enzyme H418E, in 30 mM TES buffer, 0.01 mM Mg2+, 150 mM Na+, pH 7.0 at 25°C
5.5
4-nitrophenyl beta-D-galactopyranoside
mutant enzyme H418N, in 30 mM TES buffer, 0.01 mM Mg2+, 150 mM Na+, pH 7.0 at 25°C
10.3
4-nitrophenyl beta-D-galactopyranoside
mutant enzyme H418F, in 30 mM TES buffer, 10 mM Mg2+, 150 mM Na+, pH 7.0 at 25°C
620
4-nitrophenyl beta-D-galactopyranoside
wild type enzyme, in 30 mM TES buffer, 145 mM NaCl, pH 7.0 at 25°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.8
benzyl 3-deoxy-4-S-(beta-D-galactopyranosyl)-4-thio-beta-D-erythro-pentopyranoside
-
in sodium phosphate buffer (100 mM, pH 7.3, MgCl2 1.2 mM, beta-mercaptoethanol 100 mM)
0.032
benzyl 3-deoxy-4-S-(beta-D-galactopyranosyl)-4-thio-beta-D-threo-pentopyranoside
-
in sodium phosphate buffer (100 mM, pH 7.3, MgCl2 1.2 mM, beta-mercaptoethanol 100 mM)
1
2-amino-2-deoxy-D-galactopyranose
wild type enzyme, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
1.3
2-amino-2-deoxy-D-galactopyranose
mutant enzyme H418E, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
1.4
2-amino-2-deoxy-D-galactopyranose
mutant enzyme H418N, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
0.025
2-nitrophenyl 1-thio-beta-D-galactopyranoside
wild type enzyme, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
4.2
2-nitrophenyl 1-thio-beta-D-galactopyranoside
mutant enzyme H418N, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
8.4
2-nitrophenyl 1-thio-beta-D-galactopyranoside
mutant enzyme H418E, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
0.002
2-phenylethyl 1-thio-beta-D-galactopyranoside
wild type enzyme, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
0.08
2-phenylethyl 1-thio-beta-D-galactopyranoside
mutant enzyme H418N, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
10
2-phenylethyl 1-thio-beta-D-galactopyranoside
mutant enzyme H418E, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
0.4
D-galactonolactone
mutant enzyme H418E, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
0.4
D-galactonolactone
mutant enzyme H418N, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
0.64
D-galactonolactone
wild type enzyme, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
3.3
D-galactose
mutant enzyme H418N, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
3.4
D-galactose
mutant enzyme H418E, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
21
D-galactose
wild type enzyme, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
0.08
isopropyl-thio-beta-D-galactopyranoside
wild type enzyme, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
5
isopropyl-thio-beta-D-galactopyranoside
mutant enzyme H418N, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
85
isopropyl-thio-beta-D-galactopyranoside
mutant enzyme H418E, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
2 - 5
L-arabinose
mutant enzyme H418N, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
130
L-arabinose
mutant enzyme H418E, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
220
L-arabinose
wild type enzyme, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
0.28
L-ribose
wild type enzyme, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
0.68
L-ribose
mutant enzyme H418N, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
1.4
L-ribose
mutant enzyme H418E, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
0.9
lactose
wild type enzyme, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
5.1
lactose
mutant enzyme H418N, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
13
lactose
mutant enzyme H418E, in 30 mM TES buffer, 145 mM NaCl, 0.01 mM Mg2+, pH 7.0 at 25°C
0.3
o-nitrophenyl-beta-D-galactoside
-
37°C, in 50 mM imidazole/HCl buffer containing 1 mM MgCl2, 145 mM NaCl and 4 mM dithiothreitol at pH 7.0
0.4
o-nitrophenyl-beta-D-galactoside
-
37°C, solvent: 2-methyoxyethyl acetate, 50% v/v organic solvent-buffer mixture
0.61
o-nitrophenyl-beta-D-galactoside
-
37°C, solvent: acetone, 50% v/v organic solvent-buffer mixture
0.83
o-nitrophenyl-beta-D-galactoside
-
37°C, solvent: trimethyl phosphate, 50% v/v organic solvent-buffer mixture
1.04
o-nitrophenyl-beta-D-galactoside
-
37°C, solvent: monoglyme, 50% v/v organic solvent-buffer mixture
1.21
o-nitrophenyl-beta-D-galactoside
-
37°C, solvent: acetonitrile, 50% v/v organic solvent-buffer mixture
1.44
o-nitrophenyl-beta-D-galactoside
-
37°C, solvent: diethyleneglycol diethyl ether, 50% v/v organic solvent-buffer mixture
1.67
o-nitrophenyl-beta-D-galactoside
-
37°C, solvent: tetraglyme, 50% v/v organic solvent-buffer mixture
1.71
o-nitrophenyl-beta-D-galactoside
-
37°C, solvent: diglyme, 50% v/v organic solvent-buffer mixture
1.84
o-nitrophenyl-beta-D-galactoside
-
37°C, solvent: triethyl phosphate, 50% v/v organic solvent-buffer mixture
3.38
o-nitrophenyl-beta-D-galactoside
-
37°C, solvent: triglyme, 50% v/v organic solvent-buffer mixture
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
-
a method that allows measurement of catalytic activity of a site in a preparation containing inactive protein wether homologous or foreign
additional information
-
beta-galactosidase from strains 8677 and 35321 demonstrate that the relative activities of the enzyme for the different substrates differ between the strains, assay method optimizationusing different substrates, overview
additional information
-
electrophoretic mobility and catalytic activity of individual molecules of enzyme are reproducible for each molecule but different for individual molecules. There is no relationship between the observed activities and electrophoretic motilities
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.6
6.8
-
hydrolysis of o-nitrophenyl-beta-D-galactoside, in presence of 2 mM Mg2+
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
35
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
116000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
tetramer
additional information
additional information
-
electrophoretic mobility and catalytic activity of individual molecules of enzyme are reproducible for each molecule but different for individual molecules. The observed ranges in electrophoretic motility are similar for different experimental protocols. There is no relationship between the observed activities and electrophoretic motilities
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
E416Q
the mutant enzyme does not have a Mg2+ bound at the active site even at high Mg2+ concentrations (200 mM), low catalytic activity and the pH profile is very different from that of the native enzyme
E416V
the mutant enzyme does not have a Mg2+ bound at the active site even at high Mg2+ concentrations (200 mM), low catalytic activity and the pH profile is very different from that of the native enzyme
H418F
the mutant shows low catalytic activity, the variant has reduced stability, binds Mg2+ weakly and cannot be crystallized
N460T
analysis of rate constants and activation thermodynamics to demonstrate the valuable mechanistic details of enzymes that can be obtained from studies of this type
additional information
additional information
engineering the Escherichia coli beta-galactosidase for the screening of antiviral protease inhibitors. Identification of a new accomodation site between amino acids 581 and 582, in a solvent-exposed and flexible beta-turn of domain III. the replacement of the model peptide reproducing the matrix-capsid (p17/p24) gag cleavage sequence renders a highly active and efficiently digested chimeric construct. The use of this insertion site, that increases the cleavage potential of this reporter enzyme, can improve the sensitivity and dynamic range of the antiviral drug assay
additional information
-
engineering the Escherichia coli beta-galactosidase for the screening of antiviral protease inhibitors. Identification of a new accomodation site between amino acids 581 and 582, in a solvent-exposed and flexible beta-turn of domain III. the replacement of the model peptide reproducing the matrix-capsid (p17/p24) gag cleavage sequence renders a highly active and efficiently digested chimeric construct. The use of this insertion site, that increases the cleavage potential of this reporter enzyme, can improve the sensitivity and dynamic range of the antiviral drug assay
additional information
-
a peptide of 27 amino acids, representing the G-H loop amino acid sequence of foot-and mouth disease virus VP1 protein is inserted into different sites of protein lACZ, a pseudo-wild-type beta-galactosidase lacking the 8 N-terminal amino acids
additional information
-
modulation of the catalytic properties of the enzyme using different supports and immobilization strategies, e.g. bearing glyoxyl, epoxy, or BrCN groups or by glutaraldehyde crosslinking on matrices containing primary amino groups, different derivatives exhibit very different synthetic activity/hydrolytic activity ratios in transglycosylation reactions strongly depending on the experimental conditions, overview
additional information
-
expression in Escherichia coli under control of araBD promoter. The addition of D-fucose causes an improvement in specific beta-galactosidase production, although beta-galactosidase is produced as an inclusion body. The addition of D-fucose after induction leads to an increase in the specific activity of beta-galactosidase inclusion bodies and causes a changes in the structure of beta-galactosidase inclusion bodies, with higher enzyme activity
additional information
-
single enzyme molecule assays performed on the native enzyme as well as recombinant enzyme with and without His-tag reveal significant differences in the average combined turnover number indicating that in vivo and in vitro produced enzymes are not identical and the presence of a C-terminal His-tag may alter the activity of beta-galactosidase. Average combined turnover numbers of 35800 per min and 31700 per min are obtained respectively for the in vitro synthesized enzyme from strain 35321 with the absence and presence of a C-terminal His6-tag
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4 - 10
-
the enzymatic activity of encapsulated beta-Gal remains constant after pre-incubation at pH 4-10. However, while at pH 2 the soluble enzyme is completely inactivated, encapsulated beta-Gal retains 40% of its maximum activity
703289
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37 - 75
-
soluble bet-Gal is completely inactivated when it is pre-incubated at 75°C, whereas encapsulated bet-Gal retains above 90% of its maximum activity, even after being reused, preincubation at 37°C does not affect the enzymatic activity
55
additional information
-
enzyme becomes remarkably heat labile in the presence of fructose 1,6-diphosphate
55
-
40 min, in presence of 0.1 M mercaptoethanol and 0.01 MgCl2, 50% loss of activity. Without mercaptoethanol complete loss of activity after 10 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
about 40% of the initial enzymatic activity is recovered after beta-Gal encapsulation in the silica matrix
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
2-methyoxyethyl acetate
-
37°C, 50% v/v organic solvent-buffer mixture, 5 min, about 70% loss of activity
Acetone
-
37°C, 50% v/v organic solvent-buffer mixture, 5 min, about 60% loss of activity
acetonitrile
-
37°C, 50% v/v organic solvent-buffer mixture, 5 min, about 60% loss of activity
diethyleneglycol diethyl ether
-
37°C, 50% v/v organic solvent-buffer mixture, 5 min,about 60% loss of activity
diglyme
-
37°C, 50% v/v organic solvent-buffer mixture, 5 min, about 60% loss of activity
monoglyme
-
37°C, 50% v/v organic solvent-buffer mixture, 5 min, about 50% loss of activity
tetraglyme
-
37°C, 50% v/v organic solvent-buffer mixture, 5 min, about 40% loss of activity
triethyl phosphate
-
37°C, 50% v/v organic solvent-buffer mixture, 5 min, about 40% loss of activity
triglyme
-
37°C, 50% v/v organic solvent-buffer mixture, 5 min, about 75% loss of activity
trimethyl phosphate
-
37°C, 50% v/v organic solvent-buffer mixture, 5 min, about 60% loss of activity
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
beta-galactosidase activity in 80% v/v buffer tetraglyme after 11 h is 9% of the initial activity of the enzyme in 50 mM imidazole/HCl buffer containing 1 mM MgCl2, 145 mM NaCl and 4 mM dithiothreitol at pH 7.0. The recovered activity of the enzyme, measured after dialysis is approximately 45% of the initial activity
-
refolding buffer (pH 8.0) contains 10 mM MgCl2, 0.01% Tween 20, 2.5 mM dithiothreitol, and 100 mM Tris-HCl. In a binary system using only L-Arg-HCl the refolding yield of beta-gal increases with increasing concentration up to 0.2 M, in a ternary system using both 0.2 M L-Arg-HCl and another salt, the refolding yield increases up to 1.5fold higher than that in the binary system
-
renaturation of the urea-denatured heavy isoenzymes results in the formation of tetramers but not the heavier forms
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
biotechnology
-
single enzyme molecule assays performed on the native enzyme as well as recombinant enzyme with and without His-tag reveal significant differences in the average combined turnover numbers indicating that in vivo and in vitro produced enzymes are not identical and the presence of a C-terminal His-tag may alter the activity of beta-galactosidase
environmental protection
-
the presence of coliforms in polluted water is determined enzymatically in situ by directly monitoring the activity of B-GAL through the hydrolysis of the yellow chromogenic subtrate, chlorophenol red beta-D-galactopyranoside, which produces a red chlorophenol red product, assay evaluation, overview
synthesis
analysis
-
comparison of commercially available kits to assess water quality and evaluation of their ability to detect Escherichia coli. Chromocult, MI agar, Readycult, and Colilert detect beta-glucuronidase production from respectively 79.9, 79.9, 81.1, and 51.4% of the 74 Escherichia coli strains tested. These four methods detect beta-galactosidase production from respectively 85.1, 73.8, 84.1, and 84.1% of the total coliform strains tested. The high level of false-negative results for Escherichia coli recognition obtained by all four methods suggests that they may not be appropriate for identification of presumptive Escherichia coli strains
analysis
-
electrophoretic mobility and catalytic activity of individual molecules of enzyme are reproducible for each molecule but different for individual molecules. The observed ranges in electrophoretic motility are similar for different experimental protocols. There is no relationship between the observed activities and electrophoretic motilities
analysis
-
precise and reliable detection of Escherichia coli strains for differentiation from biochemically and phylogenetically related bacteria. Method is based on polymerase chain reaction, in which four genes coding for lactose permease, cytochrome bd complex, beta-D-glucuronidase, and beta-D-galactosidase, serve as target DNA sequences
synthesis
-
simple and inexpensive method for synthesizing (2R)-glycerol-O-D-beta-galactopyranoside by utilization of the transgalalactosylating properties of beta-galactosidase and the chloroform solubility of a derivative of (2R)-glycerol-O-D-beta-galactopyranoside that is formed by the transfer of galactose onto isopropylidene glycerol
synthesis
-
expression in Escherichia coli under control of araBD promoter. The addition of D-fucose causes an improvement in specific beta-galactosidase production, although beta-galactosidase is produced as an inclusion body. The addition of D-fucose after induction leads to an increase in the specific activity of beta-galactosidase inclusion bodies and causes a changes in the structure of beta-galactosidase inclusion bodies, with higher enzyme activity
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Crueger, A.; Crueger, W.
Carbohydrates
Biotechnology (Kieslich, K. , ed. ) Verlag Chemie, Weinheim
6A
421-457
1984
Aspergillus niger, Aspergillus oryzae, Geobacillus stearothermophilus, Niallia circulans, Weizmannia coagulans, Saccharomyces cerevisiae, Caldariella acidophila, Kluyveromyces marxianus, Thermothelomyces heterothallicus, Curvularia inaequalis, Escherichia coli, Fusarium verticillioides, Trichocladium griseum, Thermomyces lanuginosus, Kluyveromyces lactis, Kluyveromyces sp., Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus sp. 'thermophilus', Leuconostoc citrovorum, Umbelopsis vinacea, Rhizomucor miehei, Rhizomucor pusillus, Penicillium multicolor, Sinorhizobium meliloti, Saccharomyces anamensis, Mycothermus thermophilus, Sporotrichum sp., Streptomyces coelicolor, Zygosaccharomyces lactis, Geobacillus stearothermophilus HRI
-
Wallenfels, K.; Weil, R.
beta-Galactosidase
The Enzymes, 3rd Ed. (Boyer, P. D. , ed. )
7
617-663
1972
Enterobacter cloacae, Aeromonas caviae, Priestia megaterium, Bacillus subtilis, Bos taurus, Streptococcus pneumoniae, Escherichia coli, Helix pomatia, Ovis aries, Homo sapiens, Kluyveromyces lactis, Lactococcus lactis, Paracolobactrum aerogenoides, Rattus norvegicus, Saccharomyces fragilis, Shigella dysenteriae, Hamamotoa singularis, Tritrichomonas suis, Escherichia coli ML 309, Escherichia coli ML 35
-
Wallenfels, K.; Malhotra, O.M.
beta-Galactosidase
The Enzymes, 2nd Ed (Boyer, P. D. , Lardy, H. , Myrbck, K. , eds. )
4
409-430
1960
Klebsiella aerogenes, Aspergillus flavus, Aspergillus niger, Aspergillus oryzae, Bos taurus, Canis lupus familiaris, Capra hircus, Coffea sp., Oryctolagus cuniculus, Escherichia coli, Felis catus, Helix pomatia, Ovis aries, Homo sapiens, Medicago sativa, Malus sp., Neurospora sp., Prunus dulcis, Prunus armeniaca, Prunus persica, Saccharomyces fragilis, Shigella sonnei, Escherichia coli K 12, Escherichia coli ML 308, Escherichia coli ML 309
-
Steers, E.; Cuatrecasas, P.
beta-Galactosidase
Methods Enzymol.
34
350-358
1974
Priestia megaterium, Escherichia coli
Boos, W.
Synthesis of (2R)-glycerol-o-beta-D-galactopyranoside by beta-galactosidase
Methods Enzymol.
89
59-64
1982
Escherichia coli
Marchesi, S.L.; Steers, E.; Shifrin, S.
Purification and characterization of the multiple forms of beta-galactosidase of Escherichia coli
Biochim. Biophys. Acta
181
20-34
1969
Escherichia coli
Feliu, J.X.; Villaverde, A.
Engineering of solvent-exposed loops in Escherichia coli beta-galactosidase
FEBS Lett.
434
23-27
1998
Escherichia coli
Jacobson, R.H.; Matthews, B.W.
Crystallization of beta-galactosidase from Escherichia coli
J. Mol. Biol.
223
1177-1182
1992
Escherichia coli
Vera, A.; Aris, A.; Daura, X.; Martinez, M.A.; Villaverde, A.
Engineering the E. coli beta-galactosidase for the screening of antiviral protease inhibitors
Biochem. Biophys. Res. Commun.
329
453-456
2005
Escherichia coli (P00722), Escherichia coli
Yoon, J.H.; Mckenzie, D.
A comparison of the activities of three beta-galactosidases in aqueous-organic solvent mixtures
Enzyme Microb. Technol.
36
439-446
2005
Aspergillus oryzae, Kluyveromyces marxianus, Escherichia coli
-
Wutor, V.C.; Togo, C.A.; Pletschke, B.I.
The effect of physico-chemical parameters and chemical compounds on the activity of beta-D-galactosidase (B-GAL), a marker enzyme for indicator microorganisms in water
Chemosphere
68
622-627
2007
Escherichia coli
Pessela, B.C.; Dellamora-Ortiz, G.; Betancor, L.; Fuentes, M.; Guisan, J.M.; Fernandez-Lafuente, R.
Modulation of the catalytic properties of multimeric beta-galactosidase from E. coli by using different immobilization protocols
Enzyme Microb. Technol.
40
310-315
2007
Escherichia coli
-
Nichols, E.R.; Gavina, J.M.; McLeod, R.G.; Craig, D.B.
Single molecule assays of beta-galactosidase from two wild-type strains of E. coli: effects of protease inhibitors on microheterogeneity and different relative activities with differing substrates
Protein J.
26
95-105
2007
Escherichia coli
Pilipenko, O.S.; Atyaksheva, L.F.; Poltorak, O.M.; Chukhrai, E.S.
Dissociation and catalytic activity of oligomer forms of beta-galactosidases
Russ. J. Phys. Chem. A
81
990-994
2007
Bos taurus, Kluyveromyces marxianus, Escherichia coli, Penicillium canescens, Penicillium canescens F-178
-
Nichols, E.R.; Craig, D.B.
Measurement of the differences in electrophoretic mobilities of individual molecules of E. coli beta-galactosidase provides insight into structural differences which underlie enzyme microheterogeneity
Electrophoresis
29
4257-4269
2008
Escherichia coli
Horakova, K.; Mlejnkova, H.; Mlejnek, P.
Specific detection of Escherichia coli isolated from water samples using polymerase chain reaction targeting four genes: cytochrome bd complex, lactose permease, beta-D-glucuronidase, and beta-D-galactosidase
J. Appl. Microbiol.
105
970-976
2008
Escherichia coli
Jung, K.H.
Enhanced enzyme activities of inclusion bodies of recombinant beta-galactosidase via the addition of inducer analog after L-arabinose induction in the araBAD promoter system of Escherichia coli
J. Microbiol. Biotechnol.
18
434-442
2008
Escherichia coli
Maheux, A.F.; Huppe, V.; Boissinot, M.; Picard, F.J.; Bissonnette, L.; Bernier, J.L.; Bergeron, M.G.
Analytical limits of four beta-glucuronidase and beta-galactosidase-based commercial culture methods used to detect Escherichia coli and total coliforms
J. Microbiol. Methods
75
506-514
2008
Escherichia coli
Nichols, E.R.; Craig, D.B.
Single molecule assays reveal differences between in vitro and in vivo synthesized beta-galactosidase
Protein J.
27
376-383
2008
Escherichia coli
Kappelhoff, J.C.; Liu, S.Y.; Dugdale, M.L.; Dymianiw, D.L.; Linton, L.R.; Huber, R.E.
Practical considerations when using temperature to obtain rate constants and activation thermodynamics of enzymes with two catalytic steps: native and N460T-beta-alactosidase (E. coli) as Examples
Protein J.
28
96-103
2009
Escherichia coli (P00722)
Cagnoni, A.J.; Uhrig, M.L.; Varela, O.
Synthesis of pentopyranosyl-containing thiodisaccharides. Inhibitory activity against beta-glycosidases
Bioorg. Med. Chem.
17
6203-6212
2009
Escherichia coli
Crescimbeni, M.C.; Nolan, V.; Clop, P.D.; Marin, G.N.; Perillo, M.A.
Activity modulation and reusability of beta-D-galactosidase confined in sol-gel derived porous silicate glass
Colloids Surf. B Biointerfaces
76
387-396
2010
Escherichia coli
Fujimoto, A.; Hirano, A.; Shiraki, K.
Ternary system of solution additives with arginine and salt for refolding of beta-galactosidase
Protein J.
29
161-166
2010
Escherichia coli
Lo, S.; Dugdale, M.L.; Jeerh, N.; Ku, T.; Roth, N.J.; Huber, R.E.
Studies of Glu-416 variants of beta-galactosidase (E. coli) show that the active site Mg2+ is not important for structure and indicate that the main role of Mg2+ is to mediate optimization of active site chemistry
Protein J.
29
26-31
2010
Escherichia coli (P00722), Escherichia coli LMG194 (P00722)
Juers, D.H.; Rob, B.; Dugdale, M.L.; Rahimzadeh, N.; Giang, C.; Lee, M.; Matthews, B.W.; Huber, R.E.
Direct and indirect roles of His-418 in metal binding and in the activity of beta-galactosidase (E. coli)
Protein Sci.
18
1281-1292
2009
Escherichia coli (P00722)
Craig, D.B.; Haslam, A.M.; Coombs, J.M.; Nichols, E.R.
Kinetic studies of unmodified individual Escherichia coli beta-galactosidase molecules in free solution
Biochem. Cell Biol.
88
451-458
2010
Escherichia coli
Hansson, T.; Andersson, M.; Wehtje, E.; Adlercreutz, P.
Influence of water activity on the competition between beta-glycosidase-catalysed transglycosylation and hydrolysis in aqueous hexanol
Enzyme Microb. Technol.
29
527-534
2001
Escherichia coli, Saccharolobus solfataricus
-
Seddigh, S.; Darabi, M.
Comprehensive analysis of beta-galactosidase protein in plants based on Arabidopsis thaliana
Turk. J. Biol.
38
140-150
2014
Asparagus officinalis, Persea americana, Musa acuminata, Brassica oleracea, Capsicum annuum, Carica papaya, Ricinus communis, Cicer arietinum, Citrus medica, Citrus sinensis, Coffea arabica, Dianthus caryophyllus, Drosophila melanogaster, Escherichia coli, Ficus carica, Fragaria x ananassa, Glycine max, Gossypium barbadense, Gossypium hirsutum, Homo sapiens, Hordeum vulgare, Linum usitatissimum, Lupinus angustifolius, Solanum lycopersicum, Mangifera indica, Malus domestica, Medicago truncatula, Mus musculus, Narcissus pseudonarcissus, Nicotiana tabacum, Pyrus communis, Petunia x hybrida, Vigna radiata, Physcomitrium patens, Populus trichocarpa, Prunus armeniaca, Prunus persica, Raphanus sativus, Sorghum bicolor, Trifolium pratense, Triticum monococcum, Vitis vinifera, Zea mays, Diospyros kaki, Oryza sativa Japonica Group, Picea sitchensis, Hymenaea courbaril, Cucumis melo var. cantalupo, Selaginella moellendorffii, Fragaria chiloensis, Prunus salicina, Actinidia deliciosa var. deliciosa, Gossypium raimondii, Eutrema halophilum, Gossypium herbaceum subsp. africanum, Sandersonia aurantiaca, Ziziphus jujuba, Arabidopsis thaliana (F4JUE3)
-
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