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 | 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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P04066 | tissue alpha-L-fucosidase precursor | - |
Thermotoga maritima | - |
- |
- |
Thermotoga maritima | Q9WYE2 | alpha-L-fucosidase, putative | - |
Purification (Comment) | Organism |
---|---|
by Ni2+-affinity chromatography | Thermotoga maritima |
recombinant | Thermotoga maritima |
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 | 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 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 |