Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination and analysis, phylogenetic tree, cloning of the modified oter 1377 gene from plasmid pUC57 to pET-30a(+) and expression as C-terminally His6-tagged protein in Escherichia coli strain BL21(DE3) | Opitutus terrae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.1 | - |
beta-D-galactosyl-(1->4)-L-rhamnose | pH 7.0, 37°C | Opitutus terrae | |
2.3 | - |
L-rhamnose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
51 | - |
2-deoxy-D-glucose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
55 | - |
L-Mannose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
58 | - |
L-Lyxose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
330 | - |
D-glucose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
370 | - |
D-xylose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
1500 | - |
D-galactose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
80000 | - |
x * 80000, recombinant C-terminally His6-tagged Oter 1377 protein, SDS-PAGE, x * 84847, recombinant C-terminally His6-tagged Oter 1377 protein, sequence calculation | Opitutus terrae |
84847 | - |
x * 80000, recombinant C-terminally His6-tagged Oter 1377 protein, SDS-PAGE, x * 84847, recombinant C-terminally His6-tagged Oter 1377 protein, sequence calculation | Opitutus terrae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
beta-D-galactosyl-(1->4)-L-rhamnose + phosphate | Opitutus terrae | - |
L-rhamnose + alpha-D-galactose 1-phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Opitutus terrae | - |
gene oter 1377 | - |
Purification (Comment) | Organism |
---|---|
recombinant C-terminally His6-tagged Oter 1377 protein from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration | Opitutus terrae |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.17 | - |
recombinant enzyme, substrate D-galactose, reverse reaction, pH 7.0, 37°C | Opitutus terrae |
0.37 | - |
recombinant enzyme, substrate D-xylose, reverse reaction, pH 7.0, 37°C | Opitutus terrae |
0.41 | - |
recombinant enzyme, substrate 2-deoxy-D-glucose, reverse reaction, pH 7.0, 37°C | Opitutus terrae |
0.62 | - |
recombinant enzyme, substrate D-glucose, reverse reaction, pH 7.0, 37°C | Opitutus terrae |
1.4 | - |
recombinant enzyme, substrate L-lyxose, reverse reaction, pH 7.0, 37°C | Opitutus terrae |
8.4 | - |
recombinant enzyme, substrate L-mannose, reverse reaction, pH 7.0, 37°C | Opitutus terrae |
27 | - |
purified recombinant enzyme, substrate beta-D-galactosyl-(1->4)-L-rhamnose, forward reaction, pH 7.0, 37°C | Opitutus terrae |
29 | - |
recombinant enzyme, substrate L-rhamnose, reverse reaction, pH 7.0, 37°C | Opitutus terrae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-deoxy-D-glucose + alpha-D-galactose 1-phosphate | reverse (synthetic) reaction | Opitutus terrae | beta-D-galactosyl-(1->4)-2-deoxy-D-glucose + phosphate | - |
r | |
beta-D-galactosyl-(1->3)-D-glucose + phosphate | lower activity | Opitutus terrae | D-glucose + alpha-D-galactose 1-phosphate | - |
r | |
beta-D-galactosyl-(1->4)-L-rhamnose + phosphate | - |
Opitutus terrae | L-rhamnose + alpha-D-galactose 1-phosphate | - |
? | |
beta-D-galactosyl-(1->4)-L-rhamnose + phosphate | preferred substrate | Opitutus terrae | L-rhamnose + alpha-D-galactose 1-phosphate | - |
r | |
D-galactose + alpha-D-galactose 1-phosphate | reverse (synthetic) reaction | Opitutus terrae | beta-D-galactosyl-(1->4)-D-galactose + phosphate | - |
r | |
D-glucose + alpha-D-galactose 1-phosphate | reverse (synthetic) reaction | Opitutus terrae | beta-D-galactosyl-(1->4)-D-glucose + phosphate | - |
r | |
D-xylose + alpha-D-galactose 1-phosphate | reverse (synthetic) reaction | Opitutus terrae | beta-D-galactosyl-(1->4)-D-xylose + phosphate | - |
r | |
L-lyxose + alpha-D-galactose 1-phosphate | reverse (synthetic) reaction | Opitutus terrae | beta-D-galactosyl-(1->4)-L-lyxose + phosphate | - |
r | |
L-mannose + alpha-D-galactose 1-phosphate | reverse (synthetic) reaction | Opitutus terrae | beta-D-galactosyl-(1->4)-L-mannose + phosphate | - |
r | |
L-rhamnose + alpha-D-galactose 1-phosphate | reverse (synthetic) reaction | Opitutus terrae | beta-D-galactosyl-(1->4)-L-rhamnose + phosphate | - |
r | |
additional information | in the reverse reaction, the enzyme shows higher activity on L-rhamnose derivatives than on D-glucose derivatives, no activity with lactose. The enzyme only exhibits activity on Gal 1-phosphate as a donor and not on alpha-D-glucose 1-phosphate and alpha-N-acetyl-D-glucosamine 1-phosphate. Various acceptors in the presence of Gal 1-P act as a donor using a reverse (synthetic) reaction with highest activity on L-Rha.The enzyme also exhibits activity on derivatives of L-Rha, but Oter 1377 protein shows no or very poor activity on D-GlcNAc, D-GalNAc, D-Man, D-glucosamine, L-Glc, D-Fru, D-Rib, L-Fuc, L-Ara, D-Lyx, D-glucal, sucrose, or maltose, substrate specificity, overview | Opitutus terrae | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 80000, recombinant C-terminally His6-tagged Oter 1377 protein, SDS-PAGE, x * 84847, recombinant C-terminally His6-tagged Oter 1377 protein, sequence calculation | Opitutus terrae |
Synonyms | Comment | Organism |
---|---|---|
D-galactosyl-beta1->4-L-rhamnose phosphorylase | - |
Opitutus terrae |
GalRhaP | - |
Opitutus terrae |
Oter 1377 protein | - |
Opitutus terrae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
- |
Opitutus terrae |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
10 | 55 | activity range, profile, overview | Opitutus terrae |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
recombinant C-terminally His6-tagged Oter 1377 protein, 30 min, 90% activity remaining | Opitutus terrae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.7 | - |
2-deoxy-D-glucose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
14 | - |
L-Lyxose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
19 | - |
D-xylose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
33 | - |
D-glucose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
37 | - |
D-galactose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
50 | - |
L-rhamnose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
60 | - |
beta-D-galactosyl-(1->4)-L-rhamnose | recombinant enzyme, pH 7.0, 37°C, forward reaction | Opitutus terrae | |
75 | - |
L-Mannose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | 7 | - |
Opitutus terrae |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 9 | activity range, profile, overview | Opitutus terrae |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
5 | 9 | recombinant C-terminally His6-tagged Oter 1377 protein, stable at | Opitutus terrae |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the glycoside hydrolase family 112 | Opitutus terrae |
physiological function | GalRhaPOt may be involved in rhamnogalacturonan I, in pectin, metabolism | Opitutus terrae |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.024 | - |
D-galactose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
0.051 | - |
D-xylose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
0.073 | - |
2-deoxy-D-glucose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
0.099 | - |
D-glucose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
0.25 | - |
L-Lyxose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
1.4 | - |
L-Mannose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae | |
22 | - |
L-rhamnose | recombinant enzyme, pH 7.0, 37°C, reverse reaction | Opitutus terrae |