Application | Comment | Organism |
---|---|---|
synthesis | a wild-type alpha-L-rhamnosidase from Alternaria sp. L1 can synthesize rhamnose-containing chemicals through reverse hydrolysis reaction with inexpensive rhamnose as glycosyl donor | Alternaria sp. L1 |
Cloned (Comment) | Organism |
---|---|
expression of wild-type enzyme and mutant enzymes in Pichia pastoris | Alternaria sp. L1 |
Protein Variants | Comment | Organism |
---|---|---|
R548A | complete loss of reverse hydrolytic activity | Alternaria sp. L1 |
W261A | complete loss of reverse hydrolytic activity | Alternaria sp. L1 |
W261Y | mutant enzyme with improved product yield in synthesis of rhamnose-containing chemicals through reverse hydrolysis reaction with rhamnose as glycosyl donor. Reverse hydrolysis is accelerated with 43.7% in relative yield compared to the wild-type enzyme. Based on 3D structural modeling, it is supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp257. Mutant W261Y also exhibits a shift in the pH-activity profile in hydrolysis reaction, indicating that introducing of a polar residue in the active site cavity may affect the catalysis behavior of the enzyme | Alternaria sp. L1 |
W555A | complete loss of reverse hydrolytic activity | Alternaria sp. L1 |
Y302F | mutant enzyme with improved product yield in synthesis of rhamnose-containing chemicals through reverse hydrolysis reaction with rhamnose as glycosyl donor. Reverse hydrolysis is accelerated with 33.3% in relative yield compared to the wild-type enzyme | Alternaria sp. L1 |
Y316F | mutant enzyme with improved product yield in synthesis of rhamnose-containing chemicals through reverse hydrolysis reaction with rhamnose as glycosyl donor. Reverse hydrolysis is accelerated with 19.8% in relative yield compared to the wild-type enzyme | Alternaria sp. L1 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.69 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, mutant enzyme Y316F | Alternaria sp. L1 | |
1.31 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, mutant enzyme W261Y | Alternaria sp. L1 | |
1.47 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, wild-type enzyme | Alternaria sp. L1 | |
2.01 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, mutant enzyme Y302F | Alternaria sp. L1 | |
2.11 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, mutant enzyme R548A | Alternaria sp. L1 | |
3.12 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, mutant enzyme W555A | Alternaria sp. L1 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Alternaria sp. L1 | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl alpha-L-rhamnopyranoside + H2O | - |
Alternaria sp. L1 | 4-nitrophenol + alpha-L-rhamnopyranose | - |
? |
Synonyms | Comment | Organism |
---|---|---|
RhaL1 | - |
Alternaria sp. L1 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0246 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, mutant enzyme W261Y | Alternaria sp. L1 | |
0.0498 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, mutant enzyme Y316F | Alternaria sp. L1 | |
0.16 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, mutant enzyme W555A | Alternaria sp. L1 | |
0.55 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, mutant enzyme Y302F | Alternaria sp. L1 | |
1.77 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, mutant enzyme R548A | Alternaria sp. L1 | |
3.07 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, wild-type enzyme | Alternaria sp. L1 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0188 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, mutant enzyme W261Y | Alternaria sp. L1 | |
0.0505 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, mutant enzyme W555A | Alternaria sp. L1 | |
0.0722 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, mutant enzyme Y316F | Alternaria sp. L1 | |
0.27 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, mutant enzyme Y302F | Alternaria sp. L1 | |
0.84 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, mutant enzyme R548A | Alternaria sp. L1 | |
2.09 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.5, 55°C, wild-type enzyme | Alternaria sp. L1 |