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Literature summary extracted from

  • Osanjo, G.; Dion, M.; Drone, J.; Solleux, C.; Tran, V.; Rabiller, C.; Tellier, C.
    Directed evolution of the alpha-L-fucosidase from Thermotoga maritima into an alpha-L-transfucosidase (2007), Biochemistry, 46, 1022-1033.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.51 expression in Escherichia coli Thermotoga maritima
3.2.1.51 into the pET21a vector, transformed into Escherichia coli Novablue cells, into plasmid pBTac2 and expressed in Escherichia coli XL1 blue MRF' Thermotoga maritima

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.51 G226S/T264A/Y267F/L322P exhibits a higher kcat than the wild-type Thermotoga maritima
3.2.1.51 G226S/T264A/Y267F/L322P the transferase/hydrolytic kinetic ratio is 22fold higher compared to wild-type ratio Thermotoga maritima
3.2.1.51 G226S/Y237H/T264A/L322P no significant modified transglycosylation properties Thermotoga maritima
3.2.1.51 G226S/Y237H/T264A/L322P the transferase/hydrolytic kinetic ratio is 34fold higher compared to wild-type ratio Thermotoga maritima
3.2.1.51 M210V/Y237H/T264A exhibits a higher kcat than the wild-type Thermotoga maritima
3.2.1.51 M210V/Y237H/T264A the transferase/hydrolytic kinetic ratio is 10fold higher compared to wild-type ratio Thermotoga maritima
3.2.1.51 M210V/Y237H/T264A/Y267F exhibits a lower kcat than the wild-type Thermotoga maritima
3.2.1.51 M210V/Y237H/T264A/Y267F the transferase/hydrolytic kinetic ratio is 11fold higher compared to wild-type ratio Thermotoga maritima
3.2.1.51 M210V/Y237H/T264A/Y267F/L322P no significant modified transglycosylation properties Thermotoga maritima
3.2.1.51 M210V/Y237H/T264A/Y267F/L322P the transferase/hydrolytic kinetic ratio is 31.5fold higher compared to wild-type ratio Thermotoga maritima
3.2.1.51 P196L/Y237H/Y267F exhibits higher transglycosylation activity than the wild-type Thermotoga maritima
3.2.1.51 P196L/Y237H/Y267F the transferase/hydrolytic kinetic ratio is 7.5fold higher compared to wild-type ratio Thermotoga maritima
3.2.1.51 R147G/L322P exhibits higher transglycosylation activity than the wild-type Thermotoga maritima
3.2.1.51 R147G/L322P the transferase/hydrolytic kinetic ratio is 8.5fold higher compared to wild-type ratio Thermotoga maritima
3.2.1.51 T264A exhibits higher transglycosylation activity than the wild-type. Causes a reorientation of the amino acids that are in direct contact with the substrates, resulting in a better docking energy Thermotoga maritima
3.2.1.51 T264A the transferase/hydrolytic kinetic ratio is 13fold higher compared to wild-type ratio Thermotoga maritima
3.2.1.51 T264A/L322P exhibits a higher kcat than the wild-type Thermotoga maritima
3.2.1.51 T264A/L322P the transferase/hydrolytic kinetic ratio is 20fold higher compared to wild-type ratio Thermotoga maritima
3.2.1.51 T264A/Y267F/L322P causes a reorientation of the amino acids that are in direct contact with the substrates, resulting in a better docking energy Thermotoga maritima
3.2.1.51 T264A/Y267F/L322P the transferase/hydrolytic kinetic ratio is 27.5fold higher compared to wild-type ratio Thermotoga maritima
3.2.1.51 Y237H/Y267F/L322P exhibits a higher kcat than the wild-type Thermotoga maritima
3.2.1.51 Y237H/Y267F/L322P the transferase/hydrolytic kinetic ratio is 20.5fold higher compared to wild-type ratio Thermotoga maritima

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.51 0.02
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, wild-type enzyme Thermotoga maritima
3.2.1.51 0.02
-
 4-nitrophenyl alpha-L-fucoside wild-type, at 40°C Thermotoga maritima
3.2.1.51 0.06
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme T264A/Y267F/L322P Thermotoga maritima
3.2.1.51 0.06
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme M210V/Y237H/T264A Thermotoga maritima
3.2.1.51 0.06
-
 4-nitrophenyl alpha-L-fucoside mutant M210V/Y237H/T264A, at 70°C. Mutant T264A/Y267F/L322P, at 40°C Thermotoga maritima
3.2.1.51 0.065
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme T264A Thermotoga maritima
3.2.1.51 0.065
-
 4-nitrophenyl alpha-L-fucoside mutant T264A, at 40°C Thermotoga maritima
3.2.1.51 0.07
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme T264A Thermotoga maritima
3.2.1.51 0.07
-
 4-nitrophenyl alpha-L-fucoside mutant T264A, at 70°C Thermotoga maritima
3.2.1.51 0.079
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, wild-type enzyme Thermotoga maritima
3.2.1.51 0.079
-
 4-nitrophenyl alpha-L-fucoside wild-type, at 70°C Thermotoga maritima
3.2.1.51 0.08
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme G226S/Y237H/T264A/L322P Thermotoga maritima
3.2.1.51 0.08
-
 4-nitrophenyl alpha-L-fucoside mutant G226S/Y237H/T264A/L322P, at 40°C Thermotoga maritima
3.2.1.51 0.09
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme G226S/T264A/Y267F/L322P Thermotoga maritima
3.2.1.51 0.09
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme M210V/Y237H/T264A/Y267F Thermotoga maritima
3.2.1.51 0.09
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme P196L/Y237H/Y267F Thermotoga maritima
3.2.1.51 0.09
-
 4-nitrophenyl alpha-L-fucoside mutant G226S/T264A/Y267F/L322P, at 40°C Thermotoga maritima
3.2.1.51 0.09
-
 4-nitrophenyl alpha-L-fucoside mutant M210V/Y237H/T264A/Y267F, at 40°C Thermotoga maritima
3.2.1.51 0.09
-
 4-nitrophenyl alpha-L-fucoside mutant P196L/Y237H/Y267F, at 40°C Thermotoga maritima
3.2.1.51 0.1
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme M210V/Y237H/T264A Thermotoga maritima
3.2.1.51 0.1
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme G226S/Y237H/T264A/L322P4 Thermotoga maritima
3.2.1.51 0.1
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme P196L/Y237H/Y267F Thermotoga maritima
3.2.1.51 0.1
-
 4-nitrophenyl alpha-L-fucoside mutant P196L/Y237H/Y267F and mutant G226S/Y237H/T264A/L322P, at 70°C. Mutant M210V/Y237H/T264A, at 40°C Thermotoga maritima
3.2.1.51 0.13
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme R147G/L322P Thermotoga maritima
3.2.1.51 0.13
-
 4-nitrophenyl alpha-L-fucoside mutant R147G/L322P, at 40°C Thermotoga maritima
3.2.1.51 0.14
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme M210V/Y237H/T264A/Y267F/L322P Thermotoga maritima
3.2.1.51 0.14
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme M210V/Y237H/T264A/Y267F Thermotoga maritima
3.2.1.51 0.14
-
 4-nitrophenyl alpha-L-fucoside mutant M210V/Y237H/T264A/Y267F, at 70°C Thermotoga maritima
3.2.1.51 0.14
-
 4-nitrophenyl alpha-L-fucoside mutant M210V/Y237H/T264A/Y267F/L322P, at 40°C Thermotoga maritima
3.2.1.51 0.19
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme T264A/L322P Thermotoga maritima
3.2.1.51 0.19
-
 4-nitrophenyl alpha-L-fucoside mutant T264A/L322P, at 40°C Thermotoga maritima
3.2.1.51 0.23
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme Y237H/Y267F/L322P Thermotoga maritima
3.2.1.51 0.23
-
 4-nitrophenyl alpha-L-fucoside mutant Y237H/Y267F/L322P, at 70°C Thermotoga maritima
3.2.1.51 0.25
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme M210V/Y237H/T264A/Y267F/L322P Thermotoga maritima
3.2.1.51 0.25
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme T264A/L322P Thermotoga maritima
3.2.1.51 0.25
-
 4-nitrophenyl alpha-L-fucoside mutant T264A/L322P7 and mutant M210V/Y237H/T264A/Y267F/L322P, at 70°C Thermotoga maritima
3.2.1.51 0.28
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme R147G/L322P Thermotoga maritima
3.2.1.51 0.28
-
 4-nitrophenyl alpha-L-fucoside mutant R147G/L322P, at 70°C Thermotoga maritima
3.2.1.51 0.29
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme Y237H/Y267F/L322P Thermotoga maritima
3.2.1.51 0.29
-
 4-nitrophenyl alpha-L-fucoside mutant Y237H/Y267F/L322P, at 40°C Thermotoga maritima
3.2.1.51 0.3
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme G226S/T264A/Y267F/L322P Thermotoga maritima
3.2.1.51 0.3
-
 4-nitrophenyl alpha-L-fucoside mutant G226S/T264A/Y267F/L322P, at 70°C Thermotoga maritima
3.2.1.51 0.31
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme T264A/Y267F/L322P Thermotoga maritima
3.2.1.51 0.31
-
 4-nitrophenyl alpha-L-fucoside mutant T264A/Y267F/L322P, at 70°C Thermotoga maritima

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.51 Homo sapiens P04066 tissue alpha-L-fucosidase precursor
-
3.2.1.51 Thermotoga maritima
-
-
-
3.2.1.51 Thermotoga maritima Q9WYE2 alpha-L-fucosidase, putative
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.51 by Ni2+-affinity chromatography Thermotoga maritima
3.2.1.51 recombinant Thermotoga maritima

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.51 4-nitrophenyl alpha-L-fucoside + H2O
-
 Thermotoga maritima 4-nitrophenol + alpha-L-fucose
-
 ?
3.2.1.51 4-nitrophenyl-alpha-L-fucopyranoside + H2O
-
 Thermotoga maritima 4-nitrophenol + alpha-L-fucose
-
 ?
3.2.1.51 additional information the wild-type enzyme catalyzes oligosaccharide synthesis by transfer of a fucosyl residue from a 4-nitrophenyl-fucoside donor to 4-nitrophenyl-fucoside (self-condensation) with alpha-(1,3) regioselectivity or 4-nitrophenyl-galactoside (transglycosylation) with alpha-(1,2) regioselectivity at low yields (7%) Thermotoga maritima ?
-
 ?
3.2.1.51 p-nitrophenyl-alpha-L-fucoside + H2O the enzyme has low transglycosylation properties Thermotoga maritima L-fucose + p-nitrophenol
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.51 alpha-L-fucosidase I
-
 Homo sapiens
3.2.1.51 Alpha-L-fucoside fucohydrolase
-
 Homo sapiens
3.2.1.51 alphafuc
-
 Thermotoga maritima
3.2.1.51 Fuca1
-
 Homo sapiens
3.2.1.51 TMalphafuc
-
 Thermotoga maritima

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.51 0.48
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme G226S/T264A/Y267F/L322P Thermotoga maritima
3.2.1.51 0.48
-
 4-nitrophenyl alpha-L-fucoside mutant G226S/T264A/Y267F/L322P, at 40°C Thermotoga maritima
3.2.1.51 0.5
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme T264A/Y267F/L322P Thermotoga maritima
3.2.1.51 0.5
-
 4-nitrophenyl alpha-L-fucoside mutant T264A/Y267F/L322P, at 40°C Thermotoga maritima
3.2.1.51 0.73
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme M210V/Y237H/T264A/Y267F/L322P Thermotoga maritima
3.2.1.51 0.73
-
 4-nitrophenyl alpha-L-fucoside mutant M210V/Y237H/T264A/Y267F/L322P, at 40°C Thermotoga maritima
3.2.1.51 2.7
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme G226S/Y237H/T264A/L322P Thermotoga maritima
3.2.1.51 2.7
-
 4-nitrophenyl alpha-L-fucoside mutant G226S/Y237H/T264A/L322P, at 40°C Thermotoga maritima
3.2.1.51 3.6
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme T264A/L322P Thermotoga maritima
3.2.1.51 3.6
-
 4-nitrophenyl alpha-L-fucoside mutant T264A/L322P, at 40°C Thermotoga maritima
3.2.1.51 4.3
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme M210V/Y237H/T264A/Y267F Thermotoga maritima
3.2.1.51 4.3
-
 4-nitrophenyl alpha-L-fucoside mutant M210V/Y237H/T264A/Y267F, at 40°C Thermotoga maritima
3.2.1.51 5.1
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, wild-type enzyme Thermotoga maritima
3.2.1.51 5.1
-
 4-nitrophenyl alpha-L-fucoside wild-type, at 40°C Thermotoga maritima
3.2.1.51 6.4
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme G226S/T264A/Y267F/L322P Thermotoga maritima
3.2.1.51 6.4
-
 4-nitrophenyl alpha-L-fucoside mutant G226S/T264A/Y267F/L322P, at 70°C Thermotoga maritima
3.2.1.51 8.2
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme T264A/Y267F/L322P Thermotoga maritima
3.2.1.51 8.2
-
 4-nitrophenyl alpha-L-fucoside mutant T264A/Y267F/L322P, at 70°C Thermotoga maritima
3.2.1.51 14.8
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme Y237H/Y267F/L322P Thermotoga maritima
3.2.1.51 14.8
-
 4-nitrophenyl alpha-L-fucoside mutant Y237H/Y267F/L322P, at 40°C Thermotoga maritima
3.2.1.51 15
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme P196L/Y237H/Y267F Thermotoga maritima
3.2.1.51 15
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme R147G/L322P Thermotoga maritima
3.2.1.51 15
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme T264A Thermotoga maritima
3.2.1.51 15
-
 4-nitrophenyl alpha-L-fucoside mutant R147G/L322P, mutant P196L/Y237H/Y267F and mutant T264A, at 40°C Thermotoga maritima
3.2.1.51 20
-
 4-nitrophenyl-alpha-L-fucopyranoside 40°C, mutant enzyme M210V/Y237H/T264A Thermotoga maritima
3.2.1.51 20
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme M210V/Y237H/T264A/Y267F/L322P Thermotoga maritima
3.2.1.51 20
-
 4-nitrophenyl alpha-L-fucoside mutant M210V/Y237H/T264A, at 40°C Thermotoga maritima
3.2.1.51 20
-
 4-nitrophenyl alpha-L-fucoside mutant M210V/Y237H/T264A/Y267F/L322P, at 70°C Thermotoga maritima
3.2.1.51 35
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, wild-type enzyme Thermotoga maritima
3.2.1.51 35
-
 4-nitrophenyl alpha-L-fucoside wild-type, at 70°C Thermotoga maritima
3.2.1.51 39
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme G226S/Y237H/T264A/L322P4 Thermotoga maritima
3.2.1.51 39
-
 4-nitrophenyl alpha-L-fucoside mutant G226S/Y237H/T264A/L322P, at 70°C Thermotoga maritima
3.2.1.51 50
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme T264A/L322P Thermotoga maritima
3.2.1.51 50
-
 4-nitrophenyl alpha-L-fucoside mutant T264A/L322P7, at 70°C Thermotoga maritima
3.2.1.51 56
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme M210V/Y237H/T264A/Y267F Thermotoga maritima
3.2.1.51 56
-
 4-nitrophenyl alpha-L-fucoside mutant M210V/Y237H/T264A/Y267F, at 70°C Thermotoga maritima
3.2.1.51 65
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme M210V/Y237H/T264A Thermotoga maritima
3.2.1.51 65
-
 4-nitrophenyl alpha-L-fucoside mutant M210V/Y237H/T264A, at 70°C Thermotoga maritima
3.2.1.51 96
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme Y237H/Y267F/L322P Thermotoga maritima
3.2.1.51 96
-
 4-nitrophenyl alpha-L-fucoside mutant Y237H/Y267F/L322P, at 70°C Thermotoga maritima
3.2.1.51 164
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme P196L/Y237H/Y267F Thermotoga maritima
3.2.1.51 164
-
 4-nitrophenyl alpha-L-fucoside mutant P196L/Y237H/Y267F, at 70°C Thermotoga maritima
3.2.1.51 169
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme T264A Thermotoga maritima
3.2.1.51 169
-
 4-nitrophenyl alpha-L-fucoside mutant T264A, at 70°C Thermotoga maritima
3.2.1.51 198
-
 4-nitrophenyl-alpha-L-fucopyranoside 70°C, mutant enzyme R147G/L322P Thermotoga maritima
3.2.1.51 198
-
 4-nitrophenyl alpha-L-fucoside mutant R147G/L322P, at 70°C Thermotoga maritima