Information on EC 2.4.1.269 - mannosylglycerate synthase

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

EC NUMBER
COMMENTARY hide
2.4.1.269
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RECOMMENDED NAME
GeneOntology No.
mannosylglycerate synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
GDP-mannose + D-glycerate = GDP + 2-O-(alpha-D-mannopyranosyl)-D-glycerate
show the reaction diagram
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Fructose and mannose metabolism
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mannosylglycerate biosynthesis II
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SYSTEMATIC NAME
IUBMB Comments
GDP-mannose:D-glycerate 2-alpha-D-mannosyltransferase
Rhodothermus marinus can also form mannosylglycerate via a two-step pathway catalysed by EC 2.4.1.217 (mannosyl-3-phosphoglycerate synthase) and EC 3.1.3.70 (mannosyl-3-phosphoglycerate phosphatase) [1]. Depending on experimental conditions mannosylglycerate synthase is more or less specific for the GDP-mannose and D-glycerate [1,2].
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
GDP-glucose + D-glycerate
2-O-(alpha-D-glucopyranosyl)-D-glycerate + GDP
show the reaction diagram
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-
-
-
?
GDP-glucose + D-glycerate
2-O-(alpha-D-mannopyranosyl)-D-glycerate + GDP
show the reaction diagram
GDP-mannose + D-glycerate
2-O-(alpha-D-mannopyranosyl)-D-glycerate + GDP
show the reaction diagram
GDP-mannose + D-lactate
?
show the reaction diagram
GDP-mannose + glycolic acid
?
show the reaction diagram
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the kcat/KM-value for glycolic acid is 70fold lower than the value for glycolic acid at 25°C
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-
?
additional information
?
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enzyme is able to efficiently synthesize mannosylglycerate and glucosylglycerate alike
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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
GDP-glucose + D-glycerate
2-O-(alpha-D-mannopyranosyl)-D-glycerate + GDP
show the reaction diagram
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the enzyme catalyzes the formation of the stress protectant 2-O-alpha-D-mannosyl glycerate
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-
?
GDP-mannose + D-glycerate
2-O-(alpha-D-mannopyranosyl)-D-glycerate + GDP
show the reaction diagram
Q9RFR0
Rhodothermus marinus possesses two enzymatic systems for the synthesis of mannosylglycerate. The first one is a single-step pathway in which mannosylglycerate synthase catalyses the synthesis of 2-O-(alpha-D-mannopyranosyl)-D-glycerate in one-step from GDP-mannose and D-glycerate. The second system is a two-step pathway in which mannosyl-3-phosphoglycerate synthase (EC 2.4.1.217) catalyses the conversion of GDP-mannose and 3-phospho-D-glycerate into 2-O-(alpha-D-mannopyranosyl)-3-phospho-D-glycerate, which is then converted to 2-O-(alpha-D-mannopyranosyl)-D-glycerate by mannosyl-3-phosphoglycerate phosphatase (EC 3.1.3.70)
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?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
may partially substitute for Mg2+
Mn2+
may partially substitute for Mg2+
additional information
NaCl and KCl, in the concentration range of 50-500 mM, have no effect on the enzyme activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Ca2+
-
divalent cation inhibit in decreasing order: Sr2+, Ni2, Co2+, Ca2+, Mn2+, Zn2+
Co2+
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divalent cation inhibit in decreasing order: Sr2+, Ni2, Co2+, Ca2+, Mn2+, Zn2+
Mn2+
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divalent cation inhibit in decreasing order: Sr2+, Ni2, Co2+, Ca2+, Mn2+, Zn2+
Ni2+
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divalent cation inhibit in decreasing order: Sr2+, Ni2, Co2+, Ca2+, Mn2+, Zn2+
Sr2+
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divalent cation inhibit in decreasing order: Sr2+, Ni2, Co2+, Ca2+, Mn2+, Zn2+
Zn2+
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divalent cation inhibit in decreasing order: Sr2+, Ni2, Co2+, Ca2+, Mn2+, Zn2+
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0014 - 121.9
D-glycerate
0.0022 - 21.3
D-lactate
124.9 - 138.6
GDP-glucose
0.00031 - 89.4
GDP-mannose
23.7
glycolic acid
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pH 7.8, 25°C, cosubstrate: GDP-mannose
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.017 - 6.5
D-glycerate
0.0026 - 0.017
D-lactate
0.63 - 1.1
GDP-glucose
0.018 - 6.1
GDP-mannose
0.0052
glycolic acid
Rhodothermus marinus
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pH 7.8, 25°C, cosubstrate: GDP-mannose
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 7
activity remains nearly constant between pH 5.5 and 7.0
7.8
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assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
85 - 90
native and recombinant enzyme
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
75 - 95
75°C: native enzyme shows about 70% of maximal activity, recombinant enzyme shows about 40% of maximal activity, 95°C: native enzyme shows about 60% of maximal activity, recombinant enzyme shows about 70% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.8
isoelectric focusing
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45000
x * 45000, SDS-PAGE
46125
x * 46125, calculated from sequence
47900
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4 * 47900, calculated
122000
gel filtration
217200
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 45000, SDS-PAGE; x * 46125, calculated from sequence
tetramer
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4 * 47900, calculated
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging-drop method
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in complex with GDP-Man/Mg2+, GDP/Mg2+, GDP/Mn2+/malonate and lactate, to 2.42-2.9 A resolution. The guanine base of GDP is housed in a hydrophobic pocket between the side chains of Lys9 and Tyr37 on one face and the aliphatic portion of the side chain of Lys76 on the opposing face, with Gln66 accepting hydrogen bonds from the endocyclic N1 and exocyclic N2
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
65
half-life: 170 min
90
half-life: 30 min for the native enzyme, 17 min for the recombinant enzyme
additional information
thermostability is not improved by Ca2+ (5 mM), nor is it enhanced under anaerobic conditions. The properties of the native and recombinant enzyme are similar, but the recombinant form is less thermostable
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
0°C, stable for more than 2 months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D100A
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inactive mutant enzyme
D135A
very high Km value for D-glycerate and a slight increase in kcat
E166A
15-fold increase in Km for glycerate
K76A
3-4-fold increase in kcat coupled with an elevation in Km for D-glycerate and GDP-Man of 15- and 3-fold, respectively
K9A
significant increase in kcat, little effect on the Km value for GDP-Man
M229A
minor influence on catalytic performance
Q66A
substantial increases in kcat and Km for both the acceptor and donor substrates
R131A
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inactive mutant enzyme
R218A
significant increase in kcat and Km values for GDP-Man, only a modest influence on the Km value for the acceptor substrate
R73A
minor influence on catalytic performance
R73G
minor influence on catalytic performance
T139A
Km value for lactate above 100 mM, modest decrease in kcat but a 1500-fold increase in the Km values for D-glycerate
W189A
3- and 7-fold increase in kcat and Km for D-glycerate, compared with the wild-type enzyme, no influence on utilization of GDP-Man
Y220A
500-fold increase in Km for D-glycerate
Y220F
1500-fold increase in Km for D-glycerate
Y37A
significant increase in kcat, little effect on the Km value for GDP-Man