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Literature summary for 2.4.1.69 extracted from

  • Engels, L.; Elling, L.
    WbgL: A novel bacterial alpha1,2-fucosyltransferase for the synthesis of 2-fucosyllactose (2014), Glycobiology, 24, 170-178.
    View publication on PubMed
Search for more literature for 2.4.1.69

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) and JM109 (DE3) cells Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
Co2+ less than 20% residual activity at 2 mM Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.27
-
 GDP-beta-L-fucose at pH 7.6 and 30°C Escherichia coli
5.3
-
 D-lactose at pH 7.6 and 30°C Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
additional information the enzyme shows no strict requirement for divalent metal ions Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
59000
-
 x * 59000, calculated from amino acid sequence Escherichia coli
70000
-
 x * 70000, recombinant, His- and pro-protein-tagged enzyme, SDS-PAGE Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
GDP-beta-L-fucose + beta-D-galactosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->3)-beta-D-galactosyl-(1->4)-beta-D-glucosyl-(1<->1)-ceramide Escherichia coli
-
 GDP + alpha-L-fucosyl-(1->2)-beta-D-galactosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->3)-beta-D-galactosyl-(1->4)-beta-D-glucosyl-(1<->1)-ceramide
-
 ?
GDP-beta-L-fucose + beta-D-galactosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->3)-beta-D-galactosyl-(1->4)-beta-D-glucosyl-(1<->1)-ceramide Escherichia coli O126
-
 GDP + alpha-L-fucosyl-(1->2)-beta-D-galactosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->3)-beta-D-galactosyl-(1->4)-beta-D-glucosyl-(1<->1)-ceramide
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli A6M9C2
-
-
Escherichia coli O126 A6M9C2
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni2+-NTA column chromatography Escherichia coli

Storage Stability

Storage Stability Organism
-20°C, 50 mM Tris-HCl buffer pH 7.6, 2 months, 30% loss of activity Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
GDP-beta-L-fucose + beta-D-galactosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->3)-beta-D-galactosyl-(1->4)-beta-D-glucosyl-(1<->1)-ceramide
-
 Escherichia coli GDP + alpha-L-fucosyl-(1->2)-beta-D-galactosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->3)-beta-D-galactosyl-(1->4)-beta-D-glucosyl-(1<->1)-ceramide
-
 ?
GDP-beta-L-fucose + beta-D-galactosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->3)-beta-D-galactosyl-(1->4)-beta-D-glucosyl-(1<->1)-ceramide
-
 Escherichia coli O126 GDP + alpha-L-fucosyl-(1->2)-beta-D-galactosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->3)-beta-D-galactosyl-(1->4)-beta-D-glucosyl-(1<->1)-ceramide
-
 ?
GDP-beta-L-fucose + D-galactose 70% activity compared to D-lactose Escherichia coli ?
-
 ?
GDP-beta-L-fucose + D-galactose 70% activity compared to D-lactose Escherichia coli O126 ?
-
 ?
GDP-beta-L-fucose + D-lactose 100% activity Escherichia coli GDP + 2'-fucosyl-D-lactose
-
 ?
GDP-beta-L-fucose + D-lactose 100% activity Escherichia coli O126 GDP + 2'-fucosyl-D-lactose
-
 ?
GDP-beta-L-fucose + D-lactulose 125% activity compared to D-lactose Escherichia coli GDP + 2'-fucosyl-D-lactulose
-
 ?
GDP-beta-L-fucose + D-lactulose 125% activity compared to D-lactose Escherichia coli O126 GDP + 2'-fucosyl-D-lactulose
-
 ?
GDP-beta-L-fucose + Gal(beta1-3)GlcNAc 28% activity compared to D-lactose Escherichia coli ?
-
 ?
GDP-beta-L-fucose + Gal(beta1-4)GlcNAc 85% activity compared to D-lactose Escherichia coli ?
-
 ?
GDP-beta-L-fucose + Galbeta-4-nitrophenyl 28% activity compared to D-lactose Escherichia coli ?
-
 ?
additional information no activity with D-galactosamine, N-acetyl-D-galactosamine, Gal(beta1-4)Glcbeta1-Bn, Gal(beta1-4)Glcbeta1-octyl, Gal(beta1-4)[Fuc(alpha1-3)]Glc, Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)GlcNAcbeta-linker-tBoc, and Gal(alpha1-6)Glc(alpha1-2)Fru Escherichia coli ?
-
 ?
additional information no activity with D-galactosamine, N-acetyl-D-galactosamine, Gal(beta1-4)Glcbeta1-Bn, Gal(beta1-4)Glcbeta1-octyl, Gal(beta1-4)[Fuc(alpha1-3)]Glc, Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)GlcNAcbeta-linker-tBoc, and Gal(alpha1-6)Glc(alpha1-2)Fru Escherichia coli O126 ?
-
 ?

Subunits

Subunits Comment Organism
? x * 59000, calculated from amino acid sequence Escherichia coli
? x * 70000, recombinant, His- and pro-protein-tagged enzyme, SDS-PAGE Escherichia coli

Synonyms

Synonyms Comment Organism
alpha1,2-fucosyltransferase
-
 Escherichia coli
alpha1,2-FucT
-
 Escherichia coli
WbgL
-
 Escherichia coli

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
27
-
 at 27°C in 50 mM Tris-HCl buffer pH 7.6, the enzyme shows a half -life of 57 h Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.066
-
 GDP-beta-L-fucose at pH 7.6 and 30°C Escherichia coli
0.067
-
 D-lactose at pH 7.6 and 30°C Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
-
 Escherichia coli

pH Range

pH Minimum pH Maximum Comment Organism
7 8
-
 Escherichia coli

pI Value

Organism Comment pI Value Maximum pI Value
Escherichia coli calculated from amino acid sequence
-
 6.2

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.013
-
 D-lactose at pH 7.6 and 30°C Escherichia coli
0.245
-
 GDP-beta-L-fucose at pH 7.6 and 30°C Escherichia coli