Information on EC 2.7.1.6 - galactokinase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea

EC NUMBER
COMMENTARY hide
2.7.1.6
-
RECOMMENDED NAME
GeneOntology No.
galactokinase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + alpha-D-galactose = ADP + alpha-D-galactose 1-phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phospho group transfer
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Amino sugar and nucleotide sugar metabolism
-
-
D-galactose degradation I (Leloir pathway)
-
-
D-galactose degradation III
-
-
D-galactose degradation V (Leloir pathway)
-
-
degradation of hexoses
-
-
Galactose metabolism
-
-
Metabolic pathways
-
-
stachyose degradation
-
-
SYSTEMATIC NAME
IUBMB Comments
ATP:D-galactose 1-phosphotransferase
Part of the Leloir pathway for galactose metabolism. The enzymes from mammals and from the bacterium Escherichia coli have no activity with N-acetyl-alpha-D-galactosamine [4-6].
CAS REGISTRY NUMBER
COMMENTARY hide
9030-53-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
dog breed Labrador retriever
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
var. Lactis
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain 279 galactose mutant
-
-
Manually annotated by BRENDA team
strain 279 galactose mutant
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain mc2
-
-
Manually annotated by BRENDA team
strain W303alpha
-
-
Manually annotated by BRENDA team
expression in Streptococcus thermophilus
-
-
Manually annotated by BRENDA team
strains IMDOST40 and IMDOST07
TrEMBL
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain QM9414, formerly Trichoderma reesei
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
-
an AtGALK T-DNA insertion mutant (atgalk) that (a) is AtGALK transcript deficient, (b) displays no GALK activity in vegetative tissues; and (c) accumulates Gal up to 6.8 mg/g FW in vegetative tissues, in contrast to wild-type plants. By constitutively overexpressing functional AtGALK cDNA, atgalk is functionally rescued
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2'-dATP + D-galactose
2'-dADP + alpha-D-galactose 1-phosphate
show the reaction diagram
-
-
-
-
?
2-amino-2-deoxy-D-galactose + ATP
2-amino-2-deoxy-D-galactose 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
2-deoxy-D-galactose + ATP
2-deoxy-D-galactose 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
3'-dATP + D-galactose
3'-dADP + alpha-D-galactose 1-phosphate
show the reaction diagram
-
-
-
-
?
3-deoxy-D-galactose + ATP
?
show the reaction diagram
-
-
-
-
?
alpha-D-galactose + ATP
alpha-D-galactose 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
ATP + 2-amino-2-deoxy-D-galactose
ADP + 2-amino-2-deoxy-D-galactose 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + 2-deoxy-D-galactose
ADP + 2-deoxy-alpha-D-galactose 1-phosphate
show the reaction diagram
ATP + 2-deoxy-D-galactose
ADP + 2-deoxy-D-galactose 1-phosphate
show the reaction diagram
ATP + 3-deoxy-3-methyl-D-galactose
ADP + 3-deoxy-3-methyl-D-galactose 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + 4-deoxy-D-galactose
ADP + 4-deoxy-alpha-D-galactose 1-phosphate
show the reaction diagram
-
substrate only for mutants Y223F, Y223W
-
-
?
ATP + 6-deoxy-D-galactose
ADP + 2-amino-2-deoxy-D-galactose 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + 6-deoxy-D-galactose
ADP + 6-deoxy-alpha-D-galactose 1-phosphate
show the reaction diagram
ATP + alpha-D-galactose
ADP + alpha-D-galactose 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + D-galactosamine
ADP + alpha-D-galactosamine 1-phosphate
show the reaction diagram
-
substrate: D-galactosamine, specific activity: 12.5 micromol/min/mg
-
-
?
ATP + D-galactosamine
ADP + D-galactosamine 1-phosphate
show the reaction diagram
ATP + D-galactose
ADP + alpha-D-galactose 1-phosphate
show the reaction diagram
ATP + D-galacturonic acid
ADP + alpha-D-galacturonic acid 1-phosphate
show the reaction diagram
-
substrate: galacturonic acid, specific activity: 0.09 micromol/min/mg
-
-
?
ATP + D-glucose
ADP + alpha-D-glucose 1-phosphate
show the reaction diagram
ATP + D-talose
ADP + alpha-D-talose 1-phosphate
show the reaction diagram
-
substrate only for mutants Y223F, Y223W
-
-
?
ATP + galactose
ADP + alpha-D-galactose 1-phosphate
show the reaction diagram
ATP + L-altrose
ADP + L-altrose 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + L-glucose
ADP + alpha-L-glucose 1-phosphate
show the reaction diagram
-
substrate only for mutants Y223F, Y223W
-
-
?
ATP + N-acetyl-D-galactosamine
ADP + N-acetyl-alpha-D-galactosamine 1-phosphate
show the reaction diagram
-
-
-
-
?
CTP + D-galactose
CDP + alpha-D-galactose 1-phosphate
show the reaction diagram
-
substrate: CTP, specific activity: 0.551 micromol/min/mg
-
-
?
D-fucose + ATP
?
show the reaction diagram
-
-
-
-
?
D-galactosamine + ATP
?
show the reaction diagram
-
-
-
-
?
D-talose + ATP
alpha-D-talose 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
dATP + D-galactose
dADP + alpha-D-galactose 1-phosphate
show the reaction diagram
-
substrate: dATP, specific activity: 138 micromol/min/mg
-
-
?
dCTP + D-galactose
dCDP + alpha-D-galactose 1-phosphate
show the reaction diagram
-
substrate: dCTP, specific activity: 0.931 micromol/min/mg
-
-
?
deoxy-ATP + D-galactose
deoxyADP + alpha-D-galactose 1-phosphate
show the reaction diagram
-
-
-
-
?
dGTP + D-galactose
dGDP + alpha-D-galactose 1-phosphate
show the reaction diagram
-
substrate: dGTP, specific activity: 31.9 micromol/min/mg
-
-
?
dTTP + D-galactose
?
show the reaction diagram
-
substrate: dTTP, specific activity: 2.5 micromol/min/mg
-
-
?
dUTP + D-galactose
dUDP + alpha-D-galactose 1-phosphate
show the reaction diagram
-
substrate: dUTP, specific activity: 1.5 micromol/min/mg
-
-
?
GTP + D-galactose
GDP + alpha-D-galactose 1-phosphate
show the reaction diagram
-
substrate: GTP, specific activity: 28.5 micromol/min/mg
-
-
?
ITP + D-galactose
IDP + alpha-D-galactose 1-phosphate
show the reaction diagram
-
substrate: ITP, specific activity: 15.7 micromol/min/mg
-
-
?
UTP + D-galactose
UDP + alpha-D-galactose 1-phosphate
show the reaction diagram
-
substrate: UTP, specific activity: 0.52 micromol/min/mg
-
-
?
additional information
?
-
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
ATP + galactose
ADP + alpha-D-galactose 1-phosphate
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ni2+
-
divalent cation required, 15% of the activation with Mg2+
Zn2+
-
can partially replace for Mg2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3-mercaptoethanol
-
-
adenosine-5'-[gamma-thio]triphosphate
-
-
alpha-D-galactose 1-phosphate
amikacin
-
-
bromo alpha-D-galactopyranoside
-
-
D-galactose 1-phosphate
fluoro alpha-D-galactopyranoside
-
-
galactose
-
above 1 mM
galactose-1-phosphate
-
-
genistein
-
-
gentamycin
-
-
iodoacetamide
-
0.05 mM, complete inactivation
kanamycin A
-
-
Mg2+
-
free Mg2+ is a competitive inhibitor of MgATP2-
neomycin
-
-
paromycin
-
-
quercetin
-
-
tobramycin
-
-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
5 mM, 2-4fold stimulation
cysteine
-
5 mM, 2-4fold stimulation
dithiothreitol
-
5 mM, 2-4fold stimulation
glutathione
-
5 mM, 2-4fold stimulation
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.9
2-amino-deoxy-D-galactose
-
-
2.2 - 29.21
2-deoxy-D-galactose
6.4
3-deoxy-3--methyl-D-galactose
-
-
4.9
6-deoxy-D-galactose
-
-
0.698 - 1.118
alpha-D-galactose
0.0039 - 3.83
ATP
1.2
D-galactosamine
-
pH 7.3, 25C
0.1 - 37.23
D-galactose
0.038 - 49.37
D-glucose
6.04 - 6.28
L-altrose
0.17
MgATP2-
-
pH 7.8
15.76
N-acetyl-D-galactosamine
-
pH 8.0, 45C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.195
2-amino-deoxy-D-galactose
Escherichia coli
-
-
0.5 - 4.01
2-deoxy-D-galactose
0.085
3-deoxy-3-methyl-D-galactose
Escherichia coli
-
-
0.0483
6-deoxy-D-galactose
Escherichia coli
-
-
0.05 - 0.65
alpha-D-galactose
0.4 - 146
ATP
0.9 - 146
D-galactose
0.007 - 1.1
D-glucose
0.36
N-acetyl-D-galactosamine
Streptococcus pneumoniae
-
pH 8.0, 45C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0022 - 0.059
2-amino-2-deoxy-D-galactose
0.019 - 0.237
2-deoxy-D-galactose
0.024 - 0.054
3-Deoxy-D-galactose
0.064 - 0.93
alpha-D-galactose
59.5
ATP
Bifidobacterium longum subsp. infantis
-
pH 8, 37C
4
0.00057 - 0.026
D-fucose
0.013 - 0.615
D-galactosamine
0.07 - 164
D-galactose
0.023
D-glucose
Streptococcus pneumoniae
-
pH 8.0, 45C
35
0.0052 - 0.14
D-talose
0.022
N-acetyl-D-galactosamine
Streptococcus pneumoniae
-
pH 8.0, 45C
1075
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.2 - 8.5
ADP
28 - 90
alpha-D-galactose 1-phosphate
10.2
AMP
-
-
2 - 3.5
D-galactose 1-phosphate
0.0739
genistein
-
-
25
Mg2+
-
-
0.0225
quercetin
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
49.9
amikacin
Rattus norvegicus
-
-
4.4
bromo alpha-D-galactopyranoside
Rattus norvegicus
-
-
0.86
fluoro alpha-D-galactopyranoside
Rattus norvegicus
-
-
16.8
Galactose 1-phosphate
Rattus norvegicus
-
-
36.6
gentamycin
Rattus norvegicus
-
-
20
kanamycin A
Rattus norvegicus
-
-
30
neomycin
Rattus norvegicus
-
-
67.3
paromycin
Rattus norvegicus
-
-
7.2
tobramycin
Rattus norvegicus
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.07
-
R37A mutant protein, substrate: galactose
0.0781
-
substrate: ATP + 6-deoxy-D-galactose, pH 8, 37C
0.0812
-
-
0.09
-
substrate: ATP + D-galacturonic acid, pH 8, 37C
0.14
-
Y109F mutant protein, substrate: galactose
0.15
-
-
0.17
-
Y236F mutant protein, substrate: galactose
0.52
-
substrate: UTP + D-galactose, pH 8, 37C
0.551
-
substrate: CTP + D-galactose, pH 8, 37C
0.61
-
mutant Y223H
0.63
-
mutant Y223R
0.65
-
mutant Y223L
0.87
-
mutant Y223W
0.91
-
wild-type protein, substrate: galactose
0.931
-
substrate: dCTP + D-galactose, pH 8, 37C
1.14
-
Y109H mutant protein, substrate: galactose
1.5
-
substrate: dUTP + D-galactose, pH 8, 37C
1.62
-
enzyme from erythrocyte
1.737
-
enzyme from placenta
2.258
-
-
2.5
-
substrate: dTTP + D-galactose, pH 8, 37C
12.5
-
substrate: ATP + D-galactosamine, pH 8, 37C
15.7
-
substrate: ITP + D-galactose, pH 8, 37C
23.42
-
mutant Y223F
23.5
-
wild-type
28.5
-
substrate: GTP + D-galactose, pH 8, 37C
31.9
-
substrate: dGTP + D-galactose, pH 8, 37C
99.7
-
substrate: ATP + 2-deoxy-D-galactose, pH 8, 37C
137
-
substrate: ATP + D-galactose, pH 8, 37C
138
-
substrate: dATP + D-galactose, pH 8, 37C
158
-
substrate: ATP + D-galactose, pH 8, 37C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.3
-
-
7.5 - 8
-
in Tris or triethanolamine buffer
7.7 - 7.9
-
Tris-glycine-NaH2PO4 buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 8
-
pH 5.5: about 60% of maximal activity, pH 8.0: about 70% of maximal activity, potassium phosphate buffer
6.7 - 8.7
-
pH 6.7: about 45% of maximal activity, pH 8.7: about 35% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 60
-
20C: about 30% of maximal activity, 60C: about 40% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.1
-
isoelectric focusing
5.3
-
isoelectric focusing
5.8
-
isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Lactococcus lactis subsp. lactis (strain IL1403)
Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37000 - 38000
-
gel filtration
37000
-
calculatd from cDNA
40000
-
gel filtration
41700
-
gel filtration
46000
-
gel filtration, calculation from sedimentation and diffusion data
47000
-
gel filtration
50000 - 55000
-
gel filtration
53000 - 57000
-
gel filtration
55000
-
gel filtration
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
in complex with D-galactose and adenosine 5-(beta,gamma-imino)-triphosphate
-
hanging drop method of vapor diffusion, wild-type and selenomethionine-labeled galactokinase
-
hanging-drop method of vapour diffusion with ammonium sulfate as the precipitant. The crystals diffract X-rays to 2.9 A resolution on a synchroton-radiation source
-
in complex with MgADP and D-galactose, comparison with S. cerevisiae enzymes
-
seeding method in combination with the sitting-drop vapour-diffusion method using 30%(v/v) PEG 400, 1 M D-galactose, 5 mM AMP-PNP, 5% (w/v) PEG 4000, 50 mM MgCl2 and 0.1 M MES pH 6.3.
-
hanging drop vapour diffusion method using 18-25% poly(ethylene glycol) 8000 containing 200 mM LiCl buffered with 100 mM MES3 (pH 6.0) and enzyme concentrations of 15.0 mg/ml and 5 mM Mg-adenosine 5'-(beta, gamma-imido)triphosphate
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
very labile below
641715
4.5
-
37C, 10 min, about 20% loss of activity
641715
4.5 - 6.5
-
25C, 30 min, stable
641711
5.4 - 8.8
-
24 h, less than 20% loss of activity
641708
6
-
stable above
641715
7
-
stable
641706
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 30
-
only 20% of maximal activity
20 - 45
-
pH 7.3, 50 mM phosphate buffer, 10 mM 2-mercaptoethanol, 5% glycerol, 60 min, stable
30
-
10 min, no loss of activity
37
-
very little activity at 37C
40
-
pH 7.5, 10 min, stable up to
55
-
nearly inactive at 55C
65
-
over 76% of maximal activity
70
-
10 min, complete loss of activity without dithiothreitol
95
-
20 min, 2 mM dithiothreitol, 60% loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
dithiothreitol protects the enzyme better than 2-mercaptoethanol against denaturation on storage at 4C
-
overnight dialysis at pH above 9.2 or below pH 5.5 results in loss of all enzymatic activity
-
the enzyme is stabilized during the purification procedure by the inclusion of 10% glycerol, 1 mM phosphate and 20 mM 2-mercaptoethanol in all buffer solutions
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 50% loss of activity after 1 months
-
-20C, unstable in 0.01 M KH2PO4-K2HPO4, pH 7.2 buffer, 0.01 M MgCl2, 1 mM dithiothreitol, 0.2 mM ATP without glycerol
-
-30C, stable for 2 months with only 5-10% loss of activity
-
-80C, less than 20% loss of activity after 6 months
-
0C, galactokinase suspension in saturated (NH4)2SO4 retains at least 80% of its activity after 1 week
-
4C, 0.01 M KH2PO4-K2HPO4, pH 7.2 buffer, 0.01 M MgCl2, 1 mM dithiothreitol, about 40% loss of activity after 24 h
-
4C, 24 h, gradual loss of activity within 2 weeks
-
4C, 3.0 M ammonium sulfate, 10% loss of activity after 6 d
-
4C, potassium phosphate buffer, pH 7.5, more than 80% of the initial activity remains after 1 week
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
HiTrap Q column chromatography, HiTrap Phenyl FF column chromatography, and HiLoad 16/60 Superdex 75 gel filtration
-
Ni2+-nitrilotriacetic acid-agarose column chromatography
-
nickel-nitrilotriacetic acid-agarose chromatography and Resource-Q column chromatography
-
one-step purification, affinity chromatography
-
recombinant enzyme
-
two forms of galactokinase, one of which is purified to homogeneity
-
using Ni-NTA chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
expressed in Escherichia coli DH5alpha cells
-
expressed in Escherichia coli strain BL21 codon plus (DE3)-RIL
-
expressed in Escherichia coli strains JM109 and K12
-
expression in COS cells, galactokinase maps to the distal region of mouse chromosome 11
-
expression in COS7 cells
expression in Escherichia coli
gene from Escherichia coli K-12 is subcloned into pET-15b vector and overexpressed in BL21 cells
-
His-tagged, expressed in Escherichia coli BL21(DE3)
-
mutant and wild-type cDNA introduced into COS cells
-
overexpression in Escherichia coli
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
enhanced gene expression caused by a nucleotide variation within GALKI promoter (c.-22T>C mutation)
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
M173L
-
moderate activity on D-glucose, wider substrate specificity than wild-type
M173L/Y371H
-
acts on additional substrates such as 4-azido-4-deoxy-D-galactose, 6-ido-6-deoxy-D-galactose, 6-chloro-6-deoxy-D-galactose, 6-bromo-6-deoxy-D-galactose, 4-deoxy-D-galactose, 6-thio-6-deoxy-D-galactose, 6-thio-6-deoxy-D-glucose, 6-azido-6-deoxy-D-glucose
Y223F
-
activity similar to wild-type, converts additional substrates L-glucose, D-talose, 4-deoxy-D-galactose, less active on D-galactosamine than wild-type
Y223H
-
significant reduction in activity
Y223L
-
significant reduction in activity
Y223R
-
significant reduction in activity
Y223W
-
significant reduction in activity, converts additional substrates L-glucose, D-talose, 4-deoxy-D-galactose, with less efficiency than mutant Y223F, less active on D-galactosamine than wild-type
Y371H
-
wider substrate specificity than wild-type
A384P
-
mutant enzyme is not present in the soluble fraction after sonication and can not be purified
C32M
-
mutant enzyme is not present in the soluble fraction after sonication and can not be purified
D46A
-
no detectable activity
E43A
-
minor changes in kinetic parameters
E43G
-
10fold drop in turnover number
E43G/H44I
-
not soluble
G346S
-
mutant enzyme shows substantial reduction in turnover number. Lower specificity constant for galactose than wild-type enzyme
G347S
-
mutant enzyme shows substantial reduction in turnover number, increase in Km-value for galactose. Lower specificity constant for galactose than wild-type enzyme
G36R
-
mutant enzyme is not present in the soluble fraction after sonication and can not be purified
H44A
-
not soluble
H44I
-
not soluble
H44Y
-
increrase in Km-value for galactose compared to wild-type. Lower specificity constant for galactose than wild-type enzyme
P28T
-
mutant enzyme is not present in the soluble fraction after sonication and can not be purified
R256W
-
drastic reduction of activity when expressed in COS cells, missense mutation causes GALK deficiency
T288M
-
mutant enzyme is not present in the soluble fraction after sonication and can not be purified
T344M
-
drastic reduction of activity when expressed in COS cells, missense mutation causes GALK deficiency
D186A
-
residue D186 interacts with galactose, possible catalytic residue, no activity of mutant enzyme
D186E
-
residue D186 interacts with galactose, possible catalytic residue, no activity of mutant enzyme
D186N
-
residue D186 interacts with galactose, possible catalytic residue, no activity of mutant enzyme
R37A
-
residue R37 interacts with galactose
Y109F
-
residue Y109interacts with ATP
Y109H
-
residue Y109interacts with ATP
Y236A
-
residue Y236 interacts with galactose
Y236F
-
residue Y236 interacts with galactose
D62A
-
abolishes all detectable galactokinase activity but retains the ability to use D-glucose as a substrate
D62A/Y274A
-
shows no sugar kinase activity
D62C
-
shows no sugar kinase activity
D62E
-
shows no detectable sugar kinase activity
D62F
-
abolishes all detectable galactokinase activity but retains the ability to use D-glucose as a substrate
D62G
-
shows no sugar kinase activity
D62H
-
abolishes all detectable galactokinase activity but retains the ability to use D-glucose as a substrate
D62I
-
shows no sugar kinase activity
D62K
-
shows no detectable sugar kinase activity
D62L
-
abolishes all detectable galactokinase activity but retains the ability to use D-glucose as a substrate
D62M
-
shows no sugar kinase activity
D62N
-
shows no detectable sugar kinase activity
D62P
-
shows no sugar kinase activity
D62Q
-
shows no sugar kinase activity
D62R
-
shows no sugar kinase activity
D62S
-
shows no sugar kinase activity
D62T
-
shows no sugar kinase activity
D62V
-
shows no sugar kinase activity
D62W
-
shows no sugar kinase activity
D62Y
-
shows no sugar kinase activity
R53A/D217A
-
shows no sugar kinase activity
D62A
-
abolishes all detectable galactokinase activity but retains the ability to use D-glucose as a substrate
-
D62E
-
shows no detectable sugar kinase activity
-
D62G
-
shows no sugar kinase activity
-
D62M
-
shows no sugar kinase activity
-
D62N
-
shows no detectable sugar kinase activity
-
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
nutrition
-
expression of enzyme in Streptococcus thermophilus, recombinant strain grows on galactose with a generation time of 55 min, which is almost double the generation time on lactose. During growth on milk and under conditions simulating those used to produce mozzarella cheese, recombinant strain grows and produces acid more rapidly than wild-type strain
synthesis
-
enzyme mutant Y371H/M173L, substrate range is much wider than wild-type, generation of unnatural sugar 1-phosphates
Show AA Sequence (3795 entries)
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