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Literature summary for 3.2.1.51 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)

Cloned (Comment) Organism
expression in Escherichia coli Thermotoga maritima
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

Protein Variants Comment Organism
G226S/T264A/Y267F/L322P exhibits a higher kcat than the wild-type Thermotoga maritima
G226S/T264A/Y267F/L322P the transferase/hydrolytic kinetic ratio is 22fold higher compared to wild-type ratio Thermotoga maritima
G226S/Y237H/T264A/L322P no significant modified transglycosylation properties Thermotoga maritima
G226S/Y237H/T264A/L322P the transferase/hydrolytic kinetic ratio is 34fold higher compared to wild-type ratio Thermotoga maritima
M210V/Y237H/T264A exhibits a higher kcat than the wild-type Thermotoga maritima
M210V/Y237H/T264A the transferase/hydrolytic kinetic ratio is 10fold higher compared to wild-type ratio Thermotoga maritima
M210V/Y237H/T264A/Y267F exhibits a lower kcat than the wild-type Thermotoga maritima
M210V/Y237H/T264A/Y267F the transferase/hydrolytic kinetic ratio is 11fold higher compared to wild-type ratio Thermotoga maritima
M210V/Y237H/T264A/Y267F/L322P no significant modified transglycosylation properties Thermotoga maritima
M210V/Y237H/T264A/Y267F/L322P the transferase/hydrolytic kinetic ratio is 31.5fold higher compared to wild-type ratio Thermotoga maritima
P196L/Y237H/Y267F exhibits higher transglycosylation activity than the wild-type Thermotoga maritima
P196L/Y237H/Y267F the transferase/hydrolytic kinetic ratio is 7.5fold higher compared to wild-type ratio Thermotoga maritima
R147G/L322P exhibits higher transglycosylation activity than the wild-type Thermotoga maritima
R147G/L322P the transferase/hydrolytic kinetic ratio is 8.5fold higher compared to wild-type ratio Thermotoga maritima
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
T264A the transferase/hydrolytic kinetic ratio is 13fold higher compared to wild-type ratio Thermotoga maritima
T264A/L322P exhibits a higher kcat than the wild-type Thermotoga maritima
T264A/L322P the transferase/hydrolytic kinetic ratio is 20fold higher compared to wild-type ratio Thermotoga maritima
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
T264A/Y267F/L322P the transferase/hydrolytic kinetic ratio is 27.5fold higher compared to wild-type ratio Thermotoga maritima
Y237H/Y267F/L322P exhibits a higher kcat than the wild-type Thermotoga maritima
Y237H/Y267F/L322P the transferase/hydrolytic kinetic ratio is 20.5fold higher compared to wild-type ratio Thermotoga maritima

KM Value [mM]

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

Organism

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

Purification (Commentary)

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl alpha-L-fucoside + H2O
-
Thermotoga maritima 4-nitrophenol + alpha-L-fucose
-
?
4-nitrophenyl-alpha-L-fucopyranoside + H2O
-
Thermotoga maritima 4-nitrophenol + alpha-L-fucose
-
?
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 ?
-
?
p-nitrophenyl-alpha-L-fucoside + H2O the enzyme has low transglycosylation properties Thermotoga maritima L-fucose + p-nitrophenol
-
?

Synonyms

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

Turnover Number [1/s]

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