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Literature summary for 2.4.1.247 extracted from

  • Nakajima, M.; Nishimoto, M.; Kitaoka, M.
    Characterization of D-galactosyl-beta1->4-L-rhamnose phosphorylase from Opitutus terrae (2010), Enzyme Microb. Technol., 46, 315-319.
No PubMed abstract available
Search for more literature for 2.4.1.247

Cloned(Commentary)

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 [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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Opitutus terrae
-
 gene oter 1377
-

Purification (Commentary)

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 [micromol/min/mg]

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 and Products (Substrate)

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

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

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
-
 Opitutus terrae

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
10 55 activity range, profile, overview Opitutus terrae

Temperature Stability [°C]

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 [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
6 7
-
 Opitutus terrae

pH Range

pH Minimum pH Maximum Comment Organism
5 9 activity range, profile, overview Opitutus terrae

pH Stability

pH Stability pH Stability Maximum Comment Organism
5 9 recombinant C-terminally His6-tagged Oter 1377 protein, stable at Opitutus terrae

General Information

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 [mM/s]

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