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EC Tree
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
D-tagatose 6-phosphate kinase, D-tagatose-6-phosphate kinase,
Lacc , phosphotagatokinase, tagatose 6-phosphate kinase (phosphorylating),
more
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D-tagatose 6-phosphate kinase
D-tagatose-6-phosphate kinase
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tagatose 6-phosphate kinase (phosphorylating)
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D-tagatose 6-phosphate kinase
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D-tagatose 6-phosphate kinase
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Lacc
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Lacc
belongs to the pfkB family of carbohydrate kinases, which form a subset of the ribokinase superfamily
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ATP + D-tagatose 6-phosphate = ADP + D-tagatose 1,6-bisphosphate
ATP + D-tagatose 6-phosphate = ADP + D-tagatose 1,6-bisphosphate
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ATP + D-tagatose 6-phosphate = ADP + D-tagatose 1,6-bisphosphate
mechanism suggested
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phospho group transfer
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ATP:D-tagatose-6-phosphate 1-phosphotransferase
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ATP + D-fructose 6-phosphate
ADP + D-fructose 1,6-bisphosphate
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ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
CTP + D-tagatose 6-phosphate
CDP + D-tagatose 1,6-bisphosphate
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GTP + D-tagatose 6-phosphate
GDP + D-tagatose 1,6-bisphosphate
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ITP + D-tagatose 6-phosphate
IDP + D-tagatose 1,6-bisphosphate
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TTP + D-tagatose 6-phosphate
TDP + D-tagatose 1,6-bisphosphate
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UTP + D-tagatose 6-phosphate
UDP + D-tagatose 1,6-bisphosphate
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additional information
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enzyme efficient biocatalytic synthesis system is developed for the phosphorylation of D-tagatose 6-phosphate at its C1-position using the recombinant D-tagatose 6-phosphate kinase and the phosphoenolpyruvate/pyruvate kinase-system for ATP regeneration. This straightforward and scalable one-step biocatalytic synthesis of D-tagatose 1,6-diphosphate is successfully scaled up to the gram scale. Method evaluation and optimization, production analysis by NMR spectroscopy, overview
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ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
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ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
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ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
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ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
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inducible by D-galactose
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ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
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involved in the catabolism of lactose and D-galactose
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ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
second step of the D-tagatose-6-phosphate pathway which is the direct catabolic pathway for lactose
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ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
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ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
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ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
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ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
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induced by growth on lactose
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ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
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?
ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
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inducible by D-galactose
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ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
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involved in the catabolism of lactose and D-galactose
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?
ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
second step of the D-tagatose-6-phosphate pathway which is the direct catabolic pathway for lactose
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ATP + D-tagatose 6-phosphate
ADP + D-tagatose 1,6-bisphosphate
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induced by growth on lactose
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ATP
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Cd2+
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8% of the activation with Mg2+, at 1.3 mM
Co2+
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54% of the activation with Mg2+, at 1.3 mM
Mn2+
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27% of the activation with Mg2+, at 1.3 mM
Ni2+
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15% of the activation with Mg2+, at 1.3 mM
Mg2+
required
Mg2+
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absolute requirement for a divalent cation, a Mg2+ : ATP ratio of 2.0-2.5 is required for maximal activity
Mg2+
required, two ions bound to the active site when substrate and cofactor are bound
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phosphate
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at high concentrations
Li+
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Li+
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33.3 mM, 65% inhibition
Na+
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Na+
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33.3 mM, 31% inhibition
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0.15
D-fructose 6-phosphate
0.016
D-tagatose 6-phosphate
0.16
ATP
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0.15
D-fructose 6-phosphate
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0.15
D-fructose 6-phosphate
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30°C, pH 8.5
0.016
D-tagatose 6-phosphate
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0.016
D-tagatose 6-phosphate
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30°C, pH 8.5
0.4
GTP
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0.17
ITP
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0.9
UTP
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0.02
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strain AHT grown on glucose
0.03
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strain B13 grown on lactose
0.043
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strain Ingbritt grown on glucose
0.045
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strain FA1 grown on glucose
0.05
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strain OMZ65 grown on lactose
0.144
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grown on galactose
0.158
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strain Ingbritt grown on galactose
0.177
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strain AHT grown on galactose
0.212
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grown on galactose
0.24
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strain Ingbritt grown on lactose
0.247
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strain FA1 grown on lactose
0.303
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grown on galactose
0.374
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strain AHT grown on lactose
0.477
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strain FA1 grown on galactose
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7.5
assay at
7.5
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in potassium phosphate buffer
8.5
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8.5
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in glycylglycine buffer, bicine buffer and Tris buffer
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7.5 - 9.5
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pH 7.5: about 75% of maximal activity, pH 9.5: about 55% of maximal activity, glycylglycine buffer
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subsp. lactis MG1820
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UniProt
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no activity in Lactobacillus plantarum
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no activity in Streptococcus faecalis
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SwissProt
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strains: AHT, FA1, Ingbritt, B13 and OMZ65
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metabolism
D-tagatose 1,6-diphosphate is an important metabolite which can be formed via the classical tagatose 6-phosphate pathway or via the tagatose 1-phosphate pathway
physiological function
enzyme LacC catalyses the efficient biocatalytic synthesis of D-tagatose 1,6-diphosphate via phosphorylation of D-tagatose 6-phosphate
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52000
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alpha2, 2 * 52000, SDS-PAGE
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dimer
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alpha2, 2 * 52000, SDS-PAGE
dimer
gel filtration data, crystal structure analysis, two monomers associate via interactions between their lid domains, which come together to form a beta-barrel structure known as a beta-clasp
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high resolution structures of D-tagatose-6-phosphate kinase (LacC) in two crystal forms. The structures define LacC in apoform, in binary complexes with ADP or the cofactor analogue adenosine 5'-(beta,gamma-imino)triphosphate, and in a ternary complex with adenosine 5'-(beta,gamma-imino)triphosphate and D-tagatose-6-phosphate. LacC adopts a closed structure required for phosphoryl transfer only when both substrate and co-factor are bound
native and L124M, L125M double mutant crystallized by hanging drop vapor diffusion method in apoform, in binary complexes with ADP or the cofactor analogue AMP-PNP, and in a ternary complex with AMP-PNP and D-tagatose-6-phosphate, wild-type protein give crystals that diffracte only to low resolution, the SeMet double mutant protein produced much more ordered crystals
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L124M/L125M
wild-type protein give crystals that diffracte only to low resolution, the SeMet double mutant protein produced much more ordered crystals
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2-mercaptoethanol stabilizes during purification
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-20°C, 20 mM potassium phosphate buffer, pH 7.5, 20% v/v glycerol, 0.2% v/v 2-mercaptoethanol, stable for several months
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native and mutant protein from Escherichia coli
recombinant enzyme from Escherichia coli strain BL21(DE3)
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expression in Escherichia coli
gene lacC, codon-optimized, recombinant expression in Escherichia coli strain BL21(DE3)
native enzyme and L124M, L125M double mutant expressed in Escherichia coli BL21(DE3)
expression in Escherichia coli
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expression in Escherichia coli
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expression in Escherichia coli
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Van Rooijen, R.J.; van Schalkwijk, S.; de Vos, W.M.
Molecular cloning, characterization, and nucleotide sequence of the tagatose 6-phosphate pathway gene cluster of the lactose operon of Lactose operon of Lactococcus lactis
J. Biol. Chem.
266
7176-7181
1991
Lactobacillus delbrueckii subsp. lactis
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Anderson, R.L.; Bissett, D.L.
D-Tagatose-6-phosphate kinase from Staphylococcus aureus
Methods Enzymol.
90
87-91
1982
Staphylococcus aureus
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Bissett, D.L.; Anderson, R.L.
Lactose and D-galactose metabolism in Staphylococcus aureus. III: Purification and properties of D-tagatose-6-phosphate kinase
J. Biol. Chem.
255
8745-8749
1980
Staphylococcus aureus
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Hamilton, I.R.; Lebtag, H.
Lactose metabolism by Streptococcus mutans: evidence for induction of the tagatose 6-phosphate pathway
J. Bacteriol.
140
1102-1104
1979
Streptococcus mutans
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Schleifer, K.H.; Hartinger, A.; Götz, F.
Occurence of D-tagatose-6-phosphate pathway of D-galactose metabolism among Staphylococci
FEMS Microbiol. Lett.
3
9-11
1978
Staphylococcus aureus, no activity in Lactobacillus plantarum, no activity in Streptococcus faecalis, Staphylococcus epidermidis, Staphylococcus hominis
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Jagusztyn-Krynicka, E.K.; Hansen, J.B.; Crow, V.L.; Thomas, T.D.; Honeyman, A.L.; Curtiss, R.III.
Streptococcus mutans serotype c tagatose 6-phosphate pathway gene cluster
J. Bacteriol.
174
6152-6158
1992
Streptococcus mutans
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Miallau, L.; Hunter, W.N.; McSweeney, S.M.; Leonard, G.A.
Structures of Staphylococcus aureus D-tagatose-6-phosphate kinase implicate domain motions in specificity and mechanism
J. Biol. Chem.
282
19948-19957
2007
Staphylococcus aureus (P0A0B9), Staphylococcus aureus
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Schoenenberger, B.; Kind, S.; Meier, R.; Eggert, T.; Obkircher, M.; Wohlgemuth, R.
Efficient biocatalytic synthesis of D-tagatose 1,6-diphosphate by LacC-catalysed phosphorylation of D-tagatose 6-phosphate
Biocatal. Biotransform.
38
53-63
2020
Lactococcus lactis subsp. lactis (P23391)
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