Information on EC 3.2.1.85 - 6-phospho-beta-galactosidase

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The expected taxonomic range for this enzyme is: Bacteria

EC NUMBER
COMMENTARY hide
3.2.1.85
-
RECOMMENDED NAME
GeneOntology No.
6-phospho-beta-galactosidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
a 6-phospho-beta-D-galactoside + H2O = 6-phospho-D-galactose + an alcohol
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
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-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Galactose metabolism
-
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lactose and galactose degradation I
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Metabolic pathways
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metabolism of disaccharids
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SYSTEMATIC NAME
IUBMB Comments
6-phospho-beta-D-galactoside 6-phosphogalactohydrolase
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CAS REGISTRY NUMBER
COMMENTARY hide
37237-42-6
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain ATCC 824
-
-
Manually annotated by BRENDA team
gene gan1D
UniProt
Manually annotated by BRENDA team
gene gan1D
UniProt
Manually annotated by BRENDA team
strain RE 1755A, Lac+Gal+ mutant CT-1
-
-
Manually annotated by BRENDA team
Klebsiella sp. RE 1755A
strain RE 1755A, Lac+Gal+ mutant CT-1
-
-
Manually annotated by BRENDA team
strain JCM1132T and strain LA117
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-
Manually annotated by BRENDA team
strain MEP181R45, strain MEP181R50 and strain MEP181R51
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-
Manually annotated by BRENDA team
strain MEP181R55 (very low actvity), strain MEP181R56 (low activity)
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Manually annotated by BRENDA team
strain MEP181R57 (very low activity) and strain MEP181R58 (very low activity)
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-
Manually annotated by BRENDA team
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Q045H2, Q045X2
UniProt
Manually annotated by BRENDA team
JCM 1031
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-
Manually annotated by BRENDA team
strain JCM2122 (very low activity)
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-
Manually annotated by BRENDA team
strain JCM2122 (very low activity)
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-
Manually annotated by BRENDA team
strain OLL2848 (very low activity), strain MEP181R68 (very low activity), strain MEP181R69 (very low activity), strain MEP181R70 (very low activity), strain MEP181R71 (very low activity), strainj MEP181R72 (very low activity), strain MEP181R74 (low activity)
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Manually annotated by BRENDA team
strain ATCC 14201
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-nitrophenyl beta-D-galactopyranoside 6-phosphate + H2O
2-nitrophenyl beta-D-galactopyranose + phosphate
show the reaction diagram
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-
-
-
?
2-nitrophenyl-beta-D-galactopyranoside 6-phosphate + H2O
2-nitrophenol + beta-D-galactose 6-phosphate
show the reaction diagram
4-nitrophenyl 6-phospho-beta-D-galactopyranoside + H2O
4-nitrophenol + 6-phospho-beta-D-galactopyranose
show the reaction diagram
-
-
-
-
?
4-nitrophenyl 6-phospho-beta-D-glucopyranoside + H2O
4-nitrophenol + 6-phospho-beta-D-glucopyranose
show the reaction diagram
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-
-
-
?
lactose + H2O
D-glucose + D-galactose
show the reaction diagram
lactose 6-phosphate + H2O
D-glucose + D-galactose 6-phosphate
show the reaction diagram
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-
-
-
?
lactose-6-phosphate + H2O
D-glucose + beta-D-galactose-6-phosphate
show the reaction diagram
o-nitrophenyl-beta-D-galactopyranoside 6-phosphate + H2O
o-nitrophenol + beta-D-galactose 6-phosphate
show the reaction diagram
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-
-
-
?
o-nitrophenyl-beta-D-galactopyranoside 6-phosphate + H2O
o-nitrophenol + beta-D-galactose-6-phosphate
show the reaction diagram
o-nitrophenyl-beta-D-galactoside + H2O
o-nitrophenol + beta-D-galactose
show the reaction diagram
-
-
-
-
?
o-nitrophenyl-beta-D-glucopyranoside 6-phosphate
o-nitrophenol + beta-D-glucose-6-phosphate
show the reaction diagram
o-nitrophenyl-beta-D-glucopyranoside 6-phosphate + H2O
o-nitrophenol + beta-D-glucose-6-phosphate
show the reaction diagram
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-
-
-
?
p-nitrophenyl-beta-D-galactopyranoside 6-phosphate + H2O
p-nitrophenol + beta-D-galactose-6-phosphate
show the reaction diagram
p-nitrophenyl-beta-D-glucopyranoside 6-phosphate
p-nitrophenol + beta-D-glucose-6-phosphate
show the reaction diagram
p-nitrophenyl-beta-D-glucopyranoside 6-phosphate + H2O
p-nitrophenol + beta-D-glucose-6-phosphate
show the reaction diagram
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less effective than ortho-form
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-
?
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
lactose + H2O
D-glucose + D-galactose
show the reaction diagram
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existence of an alternative lactose catabolic pathway in the enzyme-deficient mutant strain IL1403 by phospho-beta-galactosidase activity exhibited by BglS, a phospho-beta-glucosidase, the lactose hydrolysis is catalyzed by both phospho-beta-galactosidase and phospho-beta-glucosidase, the latter being the main enzyme in the pathway
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-
?
lactose-6-phosphate + H2O
D-glucose + beta-D-galactose-6-phosphate
show the reaction diagram
additional information
?
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physiologic regulation of the pathway, overview
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
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78% of activity at 1 mM CaCl2
Co2+
-
14% inhibition at 5 mM
Li+
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82% of control activity at 2 mM
additional information
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not required
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,4-dinitrophenyl-2-deoxy-2-bromo-beta-D-galactopyranosyl-6-phosphate
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fluoro-, chloro- and bromo-derivatives, complete inactivation, most rapidly with fluoro-, followed by chloro- and bromo-derivatives
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2,4-dinitrophenyl-2-deoxy-2-chloro-beta-D-galactopyranosyl-6-phosphate
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fluoro-, chloro- and bromo-derivatives, complete inactivation, most rapidly with fluoro-, followed by chloro- and bromo-derivatives
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2,4-dinitrophenyl-2-deoxy-2-fluoro-beta-D-galactopyranosyl-6-phosphate
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fluoro-, chloro- and bromo-derivatives, complete inactivation, most rapidly with fluoro-, followed by chloro- and bromo-derivatives
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2-phosphoglycerate
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26% inhibition at 4.2 mM
D-fructose-1,6-diphosphate
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19% inhibition at 4.2 mM
D-fructose-6-phosphate
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20% inhibition at 4.2 mM
D-galactose-6-phosphate
D-Glucose-6-phosphate
galactose-6-phosphate
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N-ethylmaleimide
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p-chloromercuribenzoic acid
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p-hydroxymercuribenzoate
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83.4% of control activity at 1 mM, 12.2% at 2 mM
p-nitrophenyl-beta-thiogalactoside 6-phosphate
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competitive inhibition
p-nitrophenyl-beta-thioglucoside 6-phosphate
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competitive inhibition
phosphoenolpyruvate
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
dithiothreitol
Fe2+
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60% increase of activity
glutathione
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L-cysteine
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.8 - 5.5
2-nitrophenyl-beta-D-galactopyranoside 6-phosphate
0.19 - 7.3
o-nitrophenyl-beta-D-galactopyranoside 6-phosphate
0.32 - 0.57
o-nitrophenyl-beta-D-galactoside
0.04 - 9.83
o-nitrophenyl-beta-D-glucopyranoside 6-phosphate
0.09
p-nitrophenyl-6-phospho-beta-D-galactopyranose
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0.1
p-nitrophenyl-6-phospho-beta-D-glucopyranose
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0.09 - 3.1
p-nitrophenyl-beta-D-galactopyranoside 6-phosphate
0.4
p-nitrophenyl-beta-D-glucopyranoside 6 phosphate
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0.1 - 21.53
p-nitrophenyl-beta-D-glucopyranoside 6-phosphate
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
206 - 361
2-nitrophenyl-beta-D-galactopyranoside 6-phosphate
57 - 83
o-nitrophenyl-beta-D-galactopyranoside 6-phosphate
4 - 9
o-nitrophenyl-beta-D-glucopyranoside 6-phosphate
22
p-nitrophenyl-6-phospho-beta-D-galactopyranose
Lactococcus lactis
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5
p-nitrophenyl-6-phospho-beta-D-glucopyranose
Lactococcus lactis
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7 - 22
p-nitrophenyl-beta-D-galactopyranoside 6-phosphate
2 - 5
p-nitrophenyl-beta-D-glucopyranoside 6-phosphate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
56 - 93
2-nitrophenyl-beta-D-galactopyranoside 6-phosphate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.004
-
with citrate-grown cell-extracts of CT-1
0.032
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with lactose-grown cell-extracts of CT-1
0.28 - 0.37
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o-nitrophenyl-beta-D-galactopyranoside 6-phosphate as substrate
0.28
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o-nitrophenyl-beta-D-galactopyranoside 6-phosphate as substrate, maximum of activity with Streptococcus lactis strain MG1299 after growth on glucose
1.45
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p-nitrophenyl-beta-D-galactopyranoside 6-phosphate as substrate, 35 mM phosphate buffer, pH 6.8
2.44
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o-nitrophenyl-beta-D-galactopyranoside 6-phosphate as substrate, maximum of activity with Streptococcus lactis strain MG1299 after growth on lactose
20.9
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purified enzyme
21.6
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purified enzyme with o-nitrophenyl-beta-D-galactopyranoside 6-phosphate as substrate, pH 7.5, 37C
23.8
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o-nitrophenyl-beta-D-galactopyranoside 6-phosphate as substrate
28.9
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o-nitrophenyl-beta-D-galactopyranoside 6-phosphate as substrate, maximum of activity
100
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maximal activity of hybrid with protein A
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5
Q045H2, Q045X2
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6
Q045H2, Q045X2
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6.5
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most active in Tris-HCl buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 7
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at 37C for 15 min
5 - 8
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nearly constant activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
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o-nitrophenyl-beta-D-galactopyranoside 6-phosphate
45 - 52
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o-nitrophenyl-beta-D-galactopyranoside 6-phosphate as substrate
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.9
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isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
activity is induced during growth on lactose but is absent in glucose-grown cells
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35000 - 56000
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gel filtration, SDS-PAGE
40000
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strain SL1, gel filtration
46000 - 53000
53000
-
SDS-PAGE
53990
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DNA sequence analysis
55100 - 56500
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DNA sequence analysis, SDS-PAGE
110000
recombinant His-tagged enzyme, gel filtration
130000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
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1 * 50000, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant His-tagged wild-type enzyme, hanging drop vapour diffusion method, two conditions and two crystal forms: 1. mixing of 3.9-4.3 mg/ml protein with reservoir solution containing 16-18% w/v PEG 8000, 3% v/v MPD, 0.1 M imidazole, pH 6.5 in a 1:1 ratio, or 2. mixing of 4.3-4.5 mg/ml protein with reservoir solution containing 19-22% w/v PEG 8000, 3% v/v MPD, 0.1 M imidazole, pH 6.48, in a 1:1 ratio, X-ray diffraction structure determination and analysis at 1.33-1.92 A resolution, detailed three-dimensional crystal structure analysis, overview
hanging drop method
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40 - 60
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P-beta-gal II, stable below 40C, inactivated at 60C
50
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rapid thermal inactivation, about 50% of activity after 3 min with extracts of strain CT-1, after 5 min with RE1755A
55
-
50% of activity after 5 min in Tris-buffer
70
-
stable up to for 5 min
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70C, purified enzyme, at least 6 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hybrid with protein A
-
native LacG1 to homogeneity by ammonium sulfate fractionation, dialysis, anion exchange and hydroxyapatite chromatography, and ultrafiltration; recombinant His6-tagged enzyme LacG2 to homogeneity by nickel affinity and adsorption chromatography, and ultrafiltration
Q045H2, Q045X2
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, the enzyme is encoded in the galoperon, including a galactose permease and the Leloir pathway enzymes
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expressed in Escherichia coli
expressed in Escherichia coli K12 TG1
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gene gan1D, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
LacG1, phylogenetic analysis; LacG2, phylogenetic analysis, recombinant expression of His6-tagged enzyme in Lactococcus lactis strain MG1363
Q045H2, Q045X2
subcloning in Escherichia coli, expression of wild-type and mutant enzymes in Lactococcus lactis strain IBB50
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the enzyme is induced by growth on lactose, as is the whole lactose catabolic system in Lactococcus garvieae, overview
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K435V/Y437F
-
significantly reduced substrate binding and turnover
S428D/K435V/Y437F
-
no activity
W429A
-
improved specificity for glucosides
E160Q
-
1000fold reduction in activity compared to wild type
E375Q
-
complete loss of activity
G332R
-
complete loss of catalytic activity, but same behavior during purification and in affinity to antisera as wild type
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
diagnostics
-
phospho-beta-galactosidase is a reliable genetic marker for biotypes of Lactococcus garvieae
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