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Literature summary for 5.1.3.11 extracted from

  • Senoura, T.; Taguchi, H.; Ito, S.; Hamada, S.; Matsui, H.; Fukiya, S.; Yokota, A.; Watanabe, J.; Wasaki, J.; Ito, S.
    Identification of the cellobiose 2-epimerase gene in the genome of Bacteroides fragilis NCTC 9343 (2009), Biosci. Biotechnol. Biochem., 73, 400-406.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Bacteroides fragilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.7 5 cellobiose recombinant enzyme, 100 mM sodium phosphate, 20 mM cellobiose, pH 7.5, 30°C Bacteroides fragilis
6.56
-
lactose recombinant enzyme, 100 mM sodium phosphate, 20 mM lactose, pH 7.5, 30°C Bacteroides fragilis
13.8
-
cellobiose recombinant enzyme, 100 mM sodium phosphate, 20 mM cellobiose, pH 7.5, 30°C Ruminococcus albus
33
-
lactose recombinant enzyme, 100 mM sodium phosphate, 20 mM lactose, pH 7.5, 30°C Ruminococcus albus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
41100
-
determined by SDS-PAGE, the mass discrepancy putatively results from partial hydrolysis at the C-terminus during expression or in the purification steps Bacteroides fragilis
45780
-
calculated by amino acid sequence, confirmed by SDS-PAGE Bacteroides fragilis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
beta-mannobiose Bacteroides fragilis
-
4-O-beta-D-mannopyranosyl-D-glucose putative product r
cellobiose Ruminococcus albus
-
4-O-beta-D-glucopyranosyl-D-mannose
-
r
cellobiose Bacteroides fragilis
-
4-O-beta-D-glucopyranosyl-D-mannose
-
r
globotriose Bacteroides fragilis
-
O-alpha-D-galactopyranosyl-(1,4)-O-beta-D-galactopyranosyl-(1,4)-D-mannose putative product r
lactose Ruminococcus albus
-
4-O-beta-D-galactopyranosyl-D-mannose
-
r
lactose Bacteroides fragilis
-
4-O-beta-D-galactopyranosyl-D-mannose
-
r
additional information Bacteroides fragilis recombinant enzyme catalyzes a hydroxyl stereoisomerism at the C2-positions of the reducing terminal glucose and at the mannose moiety of cello-oligosaccharides: lactose, beta-mannobiose and globotriose ?
-
?

Organism

Organism UniProt Comment Textmining
Bacteroides fragilis Q5LH66
-
-
Ruminococcus albus P0DKY4
-
-

Purification (Commentary)

Purification (Comment) Organism
by anion exchange chromatography, enzyme is further purified on a hydroxyapatite column Bacteroides fragilis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
28.4
-
recombinant enzyme, substrate is lactose Ruminococcus albus
29.9
-
crude extract after expression in Escherichia coli cells Bacteroides fragilis
51
-
recombinant enzyme, substrate is cellobiose Ruminococcus albus
58.3
-
recombinant enzyme after first purification step on a DEAE Sepharose FF column Bacteroides fragilis
61.8
-
recombinant enzyme after second purification step on a hydroxyapatite column, substrate is cellobiose Bacteroides fragilis
79.7
-
recombinant enzyme after second purification step on a hydroxyapatite column, substrate is lactose Bacteroides fragilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
beta-mannobiose
-
Bacteroides fragilis 4-O-beta-D-mannopyranosyl-D-glucose putative product r
cellobiose
-
Ruminococcus albus 4-O-beta-D-glucopyranosyl-D-mannose
-
r
cellobiose
-
Bacteroides fragilis 4-O-beta-D-glucopyranosyl-D-mannose
-
r
globotriose
-
Bacteroides fragilis O-alpha-D-galactopyranosyl-(1,4)-O-beta-D-galactopyranosyl-(1,4)-D-mannose putative product r
lactose
-
Ruminococcus albus 4-O-beta-D-galactopyranosyl-D-mannose
-
r
lactose
-
Bacteroides fragilis 4-O-beta-D-galactopyranosyl-D-mannose
-
r
additional information recombinant enzyme catalyzes a hydroxyl stereoisomerism at the C2-positions of the reducing terminal glucose and at the mannose moiety of cello-oligosaccharides: lactose, beta-mannobiose and globotriose Bacteroides fragilis ?
-
?

Synonyms

Synonyms Comment Organism
cellobiose 2-epimerase
-
Ruminococcus albus
cellobiose 2-epimerase
-
Bacteroides fragilis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Bacteroides fragilis
45
-
optimum Bacteroides fragilis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
52.1
-
lactose recombinant enzyme, 100 mM sodium phosphate, 20 mM lactose, pH 7.5, 30°C Ruminococcus albus
63.8
-
cellobiose recombinant enzyme, 100 mM sodium phosphate, 20 mM cellobiose, pH 7.5, 30°C Ruminococcus albus
67.6
-
cellobiose recombinant enzyme, 100 mM sodium phosphate, 20 mM cellobiose, pH 7.5, 30°C Bacteroides fragilis
79.5
-
lactose recombinant enzyme, 100 mM sodium phosphate, 20 mM lactose, pH 7.5, 30°C Bacteroides fragilis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Bacteroides fragilis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.58
-
lactose recombinant enzyme, 100 mM sodium phosphate, 20 mM lactose, pH 7.5, 30°C Ruminococcus albus
4.62
-
cellobiose recombinant enzyme, 100 mM sodium phosphate, 20 mM cellobiose, pH 7.5, 30°C Ruminococcus albus
12.1
-
lactose recombinant enzyme, 100 mM sodium phosphate, 20 mM lactose, pH 7.5, 30°C Bacteroides fragilis
18.1
-
cellobiose recombinant enzyme, 100 mM sodium phosphate, 20 mM cellobiose, pH 7.5, 30°C Bacteroides fragilis