BRENDA - Enzyme Database show
show all sequences of 2.4.1.288

Tetrameric structure of the GlfT2 galactofuranosyltransferase reveals a scaffold for the assembly of mycobacterial arabinogalactan

Wheatley, R.W.; Zheng, R.B.; Richards, M.R.; Lowary, T.L.; Ng, K.K.; J. Biol. Chem. 287, 28132-28143 (2012)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli C41(DE3) cells
Mycobacterium tuberculosis
Crystallization (Commentary)
Crystallization
Organism
hanging drop vapor diffusion method, free enzyme is crystallized using 5-7% (w/v) PEG 4000, 100 mM sodium cacodylate, pH 6.5, 100 mM sodium acetate. UDP-bound enzyme is crystallized using 6-7% (w/v) PEG 4000, 100 mM sodium acetate, 10% (v/v) glycerol, 5 mM TCEP, at 22°C
Mycobacterium tuberculosis
Engineering
Amino acid exchange
Commentary
Organism
D372S
the mutant shows no detectable activity
Mycobacterium tuberculosis
E300S
the mutant exhibits increased Km and strongly decreased kcat values compare to the wild type enzyme
Mycobacterium tuberculosis
E371S
the mutant shows no detectable activity
Mycobacterium tuberculosis
H413S
the mutant exhibits increased Km and strongly decreased kcat values compare to the wild type enzyme
Mycobacterium tuberculosis
W399S
the mutant exhibits increased Km and strongly decreased kcat values compare to the wild type enzyme
Mycobacterium tuberculosis
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.38
-
UDP-alpha-D-galactofuranose
wild type enzyme, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.41
-
UDP-alpha-D-galactofuranose
mutant enzyme W399S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.42
-
UDP-alpha-D-galactofuranose
mutant enzyme E300S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.43
-
UDP-alpha-D-galactofuranose
mutant enzyme H413S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.6
-
beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
wild type enzyme, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
1.54
-
beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
mutant enzyme H413S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
1.78
-
beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
mutant enzyme E300S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C; mutant enzyme W399S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mn2+
Mn2+ is bound to the active site
Mycobacterium tuberculosis
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Mycobacterium tuberculosis
O53585
-
-
Purification (Commentary)
Commentary
Organism
nickel-Sepharose column chromatography and Sephacryl S-200 gel filtration
Mycobacterium tuberculosis
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
UDP-alpha-D-galactofuranose + beta-D-galactofuranosyl-(1->5)-beta-D-galactofuranosyl-(1->4)-alpha-L-rhamnopyranosyl-(1->3)-N-acetyl-alpha-D-glucosaminyl-diphospho-trans,octacis-decaprenol
the beta-(1->5)-transferase activity of the enzyme first glycosylates the galactofuranosyl residue at the nonreducing terminus of the polyprenol-bound tetrasaccharide acceptor. The resulting product is then a substrate for the beta-(1->6)-transferase activity of the enzyme
720037
Mycobacterium tuberculosis
UDP + [beta-D-galactofuranosyl-(1->5)-beta-D-galactofuranosyl-(1->6)]14-beta-D-galactofuranosyl-(1->5)-beta-D-galactofuranosyl-(1->4)-alpha-L-rhamnopyranosyl-(1->3)-N-acetyl-alpha-D-glucosaminyl-diphospho-trans,octacis-decaprenol
-
-
-
?
UDP-alpha-D-galactofuranose + beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
-
720037
Mycobacterium tuberculosis
UDP + ?
-
-
-
?
Subunits
Subunits
Commentary
Organism
homotetramer
x-ray crystallography
Mycobacterium tuberculosis
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.004
-
beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
mutant enzyme H413S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.004
-
UDP-alpha-D-galactofuranose
mutant enzyme H413S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.005
-
beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
mutant enzyme E300S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.005
-
UDP-alpha-D-galactofuranose
mutant enzyme E300S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.006
-
beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
mutant enzyme W399S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.006
-
UDP-alpha-D-galactofuranose
mutant enzyme W399S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
7.2
-
beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
wild type enzyme, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
7.2
-
UDP-alpha-D-galactofuranose
wild type enzyme, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli C41(DE3) cells
Mycobacterium tuberculosis
Crystallization (Commentary) (protein specific)
Crystallization
Organism
hanging drop vapor diffusion method, free enzyme is crystallized using 5-7% (w/v) PEG 4000, 100 mM sodium cacodylate, pH 6.5, 100 mM sodium acetate. UDP-bound enzyme is crystallized using 6-7% (w/v) PEG 4000, 100 mM sodium acetate, 10% (v/v) glycerol, 5 mM TCEP, at 22°C
Mycobacterium tuberculosis
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
D372S
the mutant shows no detectable activity
Mycobacterium tuberculosis
E300S
the mutant exhibits increased Km and strongly decreased kcat values compare to the wild type enzyme
Mycobacterium tuberculosis
E371S
the mutant shows no detectable activity
Mycobacterium tuberculosis
H413S
the mutant exhibits increased Km and strongly decreased kcat values compare to the wild type enzyme
Mycobacterium tuberculosis
W399S
the mutant exhibits increased Km and strongly decreased kcat values compare to the wild type enzyme
Mycobacterium tuberculosis
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.38
-
UDP-alpha-D-galactofuranose
wild type enzyme, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.41
-
UDP-alpha-D-galactofuranose
mutant enzyme W399S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.42
-
UDP-alpha-D-galactofuranose
mutant enzyme E300S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.43
-
UDP-alpha-D-galactofuranose
mutant enzyme H413S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.6
-
beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
wild type enzyme, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
1.54
-
beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
mutant enzyme H413S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
1.78
-
beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
mutant enzyme E300S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C; mutant enzyme W399S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mn2+
Mn2+ is bound to the active site
Mycobacterium tuberculosis
Purification (Commentary) (protein specific)
Commentary
Organism
nickel-Sepharose column chromatography and Sephacryl S-200 gel filtration
Mycobacterium tuberculosis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
UDP-alpha-D-galactofuranose + beta-D-galactofuranosyl-(1->5)-beta-D-galactofuranosyl-(1->4)-alpha-L-rhamnopyranosyl-(1->3)-N-acetyl-alpha-D-glucosaminyl-diphospho-trans,octacis-decaprenol
the beta-(1->5)-transferase activity of the enzyme first glycosylates the galactofuranosyl residue at the nonreducing terminus of the polyprenol-bound tetrasaccharide acceptor. The resulting product is then a substrate for the beta-(1->6)-transferase activity of the enzyme
720037
Mycobacterium tuberculosis
UDP + [beta-D-galactofuranosyl-(1->5)-beta-D-galactofuranosyl-(1->6)]14-beta-D-galactofuranosyl-(1->5)-beta-D-galactofuranosyl-(1->4)-alpha-L-rhamnopyranosyl-(1->3)-N-acetyl-alpha-D-glucosaminyl-diphospho-trans,octacis-decaprenol
-
-
-
?
UDP-alpha-D-galactofuranose + beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
-
720037
Mycobacterium tuberculosis
UDP + ?
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
homotetramer
x-ray crystallography
Mycobacterium tuberculosis
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.004
-
beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
mutant enzyme H413S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.004
-
UDP-alpha-D-galactofuranose
mutant enzyme H413S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.005
-
beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
mutant enzyme E300S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.005
-
UDP-alpha-D-galactofuranose
mutant enzyme E300S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.006
-
beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
mutant enzyme W399S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
0.006
-
UDP-alpha-D-galactofuranose
mutant enzyme W399S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
7.2
-
beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
wild type enzyme, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
7.2
-
UDP-alpha-D-galactofuranose
wild type enzyme, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C
Mycobacterium tuberculosis
General Information
General Information
Commentary
Organism
physiological function
GlfT2 is essential for mycobacterial cell wall biosynthesis
Mycobacterium tuberculosis
General Information (protein specific)
General Information
Commentary
Organism
physiological function
GlfT2 is essential for mycobacterial cell wall biosynthesis
Mycobacterium tuberculosis
Other publictions for EC 2.4.1.288
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
735959
Dumitrescu
Selectfluor and NFSI exo-glyca ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis ATCC 25618
Chemistry
20
15208-15215
2014
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2
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3
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2
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2
2
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736729
Li
-
Sulfonium ions as inhibitors o ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis ATCC 25618
MedChemComm
5
1130-1137
2014
-
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-
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2
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3
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2
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718920
May
A processive carbohydrate poly ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
Biochemistry
51
1148-1159
2012
-
-
-
-
7
-
1
-
-
2
-
-
-
160
-
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2
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7
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1
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2
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-
2
-
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-
-
-
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-
1
1
-
-
-
719665
Brown
Fluorosugar chain termination ...
Mycobacterium tuberculosis
J. Am. Chem. Soc.
134
6552-6555
2012
-
-
-
-
-
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-
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2
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1
1
-
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-
720037
Wheatley
Tetrameric structure of the Gl ...
Mycobacterium tuberculosis
J. Biol. Chem.
287
28132-28143
2012
-
-
1
1
5
-
-
7
-
1
-
-
-
1
-
-
1
-
-
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2
1
-
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-
8
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-
1
-
1
5
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-
7
-
1
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-
-
1
-
-
-
-
2
1
-
-
-
8
-
-
-
-
-
1
1
-
-
-
720611
Poulin
Synthetic UDP-galactofuranose ...
Mycobacterium tuberculosis
Org. Biomol. Chem.
10
4074-4087
2012
-
-
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-
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6
7
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-
1
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-
11
-
14
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7
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1
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6
1
7
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11
-
14
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7
-
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7
7
719284
Szczepina
STD-NMR studies suggest that t ...
Mycobacterium tuberculosis
ChemBioChem
10
2052-2059
2009
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2
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3
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-
720894
May
A tethering mechanism for leng ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
Proc. Natl. Acad. Sci. USA
106
11851-11856
2009
-
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1
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-
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161
-
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1
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12
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1
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1
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12
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719218
Rose
Development of a coupled spect ...
Mycobacterium tuberculosis
Carbohydr. Res.
343
2130-2139
2008
-
-
1
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-
1
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1
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2
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1
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2
3
1
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1
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1
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1
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1
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1
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2
3
1
-
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1
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-
-
-
-
-
-
-
-
719712
Belanova
Galactosyl transferases in myc ...
Mycolicibacterium smegmatis
J. Bacteriol.
190
1141-1145
2008
-
-
1
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-
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4
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3
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1
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3
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1
1
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-
719707
Mikusova
Identification of a novel gala ...
Mycobacterium tuberculosis
J. Bacteriol.
188
6592-6598
2006
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1
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3
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2
1
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1
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2
1
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1
1
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