Cloned (Comment) | Organism |
---|---|
the recombinant protein is fused to a C-terminal His-tag and expressed in Escherichia coli BL21(DE3) | Novosphingobium sp. PP1Y |
Protein Variants | Comment | Organism |
---|---|---|
D503A | no detectable activity | Novosphingobium sp. PP1Y |
D552A | kcat/KM value is 65% of the wild-type value. This reduced catalytic efficiency is mainly due to a significative variation in the turnover number, defined by the kcat value, for which a 34% decrease is observed | Novosphingobium sp. PP1Y |
D552A | mutant enzyme shows 47.9% of residual activity | Novosphingobium sp. PP1Y |
D763A | kcat/KM value is 21% of the wild-type value. A 82% decrease in kcat is retrieved for the mutant enzyme, while the KM is similar within the experimental error | Novosphingobium sp. PP1Y |
D763A | mutant enzyme shows 15.8% of residual activity | Novosphingobium sp. PP1Y |
E506A | mutant enzyme shows 0.05% of residual activity | Novosphingobium sp. PP1Y |
E644A | no detectable activity | Novosphingobium sp. PP1Y |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.031 | - |
rutin | pH 6.9, 37°C | Novosphingobium sp. PP1Y | |
0.067 | - |
hesperidin | pH 6.9, 37°C | Novosphingobium sp. PP1Y | |
0.138 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.9, 37°C, mutant enzyme D763A | Novosphingobium sp. PP1Y | |
0.15 | - |
quercitrin | pH 6.9, 37°C | Novosphingobium sp. PP1Y | |
0.157 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.9, 37°C, wild-type enzyme | Novosphingobium sp. PP1Y | |
0.159 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.9, 37°C, mutant enzyme D552A | Novosphingobium sp. PP1Y | |
0.59 | - |
naringin | pH 6.9, 37°C | Novosphingobium sp. PP1Y |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | EDTA-treated enzyme recovers its activity in the presence of divalent cations such as Mg2+ and Ca2+, but with an apparent higher efficiency when calcium is used. Almost total recovery of activity in the presence of a 3000-fold excess of Ca2+ compared to the 56% obtained with a similar excess of Mg2+. Five residues contribute to the binding of the Ca2+ ion: E593, E538, D458, S459 and E561 | Novosphingobium sp. PP1Y | |
Mg2+ | EDTA-treated enzyme recovers its activity in the presence of divalent cations such as Mg2+ and Ca2+, but with an apparent higher efficiency when calcium is used. Almost total recovery of activity in the presence of a 3000-fold excess of Ca2+ compared to the 56% obtained with a similar excess of Mg2+ | Novosphingobium sp. PP1Y |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
12000 | - |
- |
Novosphingobium sp. PP1Y |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Novosphingobium sp. PP1Y | F6IEX3 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Novosphingobium sp. PP1Y |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl alpha-L-rhamnopyranoside + H2O | - |
Novosphingobium sp. PP1Y | 4-nitrophenol + alpha-L-rhamnopyranose | - |
? | |
hesperidin + H2O | - |
Novosphingobium sp. PP1Y | hesperetin 7-O-beta-D-glucoside + alpha-L-rhamnose | - |
? | |
additional information | the enzyme is able to hydrolyze rhamnose from natural flavonoids. It hydrolyzes both alpha-1,2 and alpha-1,6 glycosidic linkages | Novosphingobium sp. PP1Y | ? | - |
? | |
naringin + H2O | - |
Novosphingobium sp. PP1Y | beta-D-glucopyranosyl-2,3-dihydro-4',5,7-trihydroxyflavone + alpha-L-rhamnose | - |
? | |
quercitrin + H2O | - |
Novosphingobium sp. PP1Y | quercetin + alpha-L-rhamnose | - |
? | |
rutin + H2O | - |
Novosphingobium sp. PP1Y | isoquercitrin + alpha-L-rhamnose | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | - |
Novosphingobium sp. PP1Y |
Synonyms | Comment | Organism |
---|---|---|
GH106 alpha-L-rhamnosidase | - |
Novosphingobium sp. PP1Y |
RHA-P | - |
Novosphingobium sp. PP1Y |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
35 | 45 | - |
Novosphingobium sp. PP1Y |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
12.2 | - |
quercitrin | pH 6.9, 37°C | Novosphingobium sp. PP1Y | |
12.7 | - |
hesperidin | pH 6.9, 37°C | Novosphingobium sp. PP1Y | |
18.2 | - |
rutin | pH 6.9, 37°C | Novosphingobium sp. PP1Y | |
126 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.9, 37°C, mutant enzyme D763A | Novosphingobium sp. PP1Y | |
214 | - |
naringin | pH 6.9, 37°C | Novosphingobium sp. PP1Y | |
458 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.9, 37°C, mutant enzyme D552A | Novosphingobium sp. PP1Y | |
692 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.9, 37°C, wild-type enzyme | Novosphingobium sp. PP1Y |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | 7.5 | - |
Novosphingobium sp. PP1Y |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
80 | - |
quercitrin | pH 6.9, 37°C | Novosphingobium sp. PP1Y | |
190 | - |
hesperidin | pH 6.9, 37°C | Novosphingobium sp. PP1Y | |
360 | - |
naringin | pH 6.9, 37°C | Novosphingobium sp. PP1Y | |
586 | - |
rutin | pH 6.9, 37°C | Novosphingobium sp. PP1Y | |
900 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.9, 37°C, mutant enzyme D763A | Novosphingobium sp. PP1Y | |
2900 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.9, 37°C, mutant enzyme D552A | Novosphingobium sp. PP1Y | |
4400 | - |
4-nitrophenyl alpha-L-rhamnopyranoside | pH 6.9, 37°C, wild-type enzyme | Novosphingobium sp. PP1Y |