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Literature summary for 2.4.1.320 extracted from

  • Nihira, T.; Suzuki, E.; Kitaoka, M.; Nishimoto, M.; Ohtsubo, K.; Nakai, H.
    Discovery of beta-1,4-D-mannosyl-N-acetyl-D-glucosamine phosphorylase involved in the metabolism of N-glycans (2013), J. Biol. Chem., 288, 27366-27374.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli DH5alpha Bacteroides thetaiotaomicron
expression in Escherichia coli Bacteroides thetaiotaomicron

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.6
-
phosphate pH 5.5, 30°C, cosubstrate: 4-O-beta-D-mannopyranosyl-N-acetyl-D-glucosamine, phosphorolytic reaction Bacteroides thetaiotaomicron
2.1
-
4-O-beta-D-mannopyranosyl-N-acetyl-D-glucosamine pH 5.5, 30°C, cosubstrate: phosphate, phosphorolytic reaction Bacteroides thetaiotaomicron
2.6
-
alpha-D-mannose 1-phosphate pH 5.5, 30°C Bacteroides thetaiotaomicron
2.6
-
alpha-D-mannose 1-phosphate pH 5.5, 30°C, cosubstrate: N-acetyl-D-glucosamine, synthetic reaction Bacteroides thetaiotaomicron
2.7
-
4-O-beta-N-acetyl-D-glucosaminyl-N-acetyl-D-glucosamine pH 5.5, 30°C Bacteroides thetaiotaomicron
3.8
-
N-acetyl-D-glucosamine pH 5.5, 30°C Bacteroides thetaiotaomicron
3.8
-
N-acetyl-D-glucosamine pH 5.5, 30°C, synthetic reaction Bacteroides thetaiotaomicron
27
-
N,N'-diacetylchitobiose pH 5.5, 30°C, synthetic reaction Bacteroides thetaiotaomicron

Localization

Localization Comment Organism GeneOntology No. Textmining
cytoplasm
-
Bacteroides thetaiotaomicron 5737
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35000
-
4 * 35000, SDS-PAGE Bacteroides thetaiotaomicron
35000
-
4 * 35000, SDS-PAGE, 4 * 37823, amino acid sequence Bacteroides thetaiotaomicron
37823
-
4 * 35000, SDS-PAGE, 4 * 37823, amino acid sequence Bacteroides thetaiotaomicron
37823
-
4 * 37823, calculated from sequence Bacteroides thetaiotaomicron
147000
-
gel filtration Bacteroides thetaiotaomicron

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4-O-beta-D-mannopyranosyl-N-acetyl-D-glucosamine + phosphate Bacteroides thetaiotaomicron the enzyme is involved in the degradation of host-derived N-glycans N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
r
4-O-beta-D-mannopyranosyl-N-acetyl-D-glucosamine + phosphate Bacteroides thetaiotaomicron VPI-5482 the enzyme is involved in the degradation of host-derived N-glycans N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
r
4-O-beta-D-mannosyl-N-acetyl-D-glucosamine + phosphate Bacteroides thetaiotaomicron
-
N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
r
4-O-beta-D-mannosyl-N-acetyl-D-glucosamine + phosphate Bacteroides thetaiotaomicron VPI-5482
-
N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
r
N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate Bacteroides thetaiotaomicron
-
4-O-beta-D-mannosyl-N-acetyl-D-glucosamine + phosphate
-
r
N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate Bacteroides thetaiotaomicron VPI-5482
-
4-O-beta-D-mannosyl-N-acetyl-D-glucosamine + phosphate
-
r

Organism

Organism UniProt Comment Textmining
Bacteroides thetaiotaomicron Q8A8X4
-
-
Bacteroides thetaiotaomicron Q8A8Y4
-
-
Bacteroides thetaiotaomicron VPI-5482 Q8A8X4
-
-
Bacteroides thetaiotaomicron VPI-5482 Q8A8Y4
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Bacteroides thetaiotaomicron

Reaction

Reaction Comment Organism Reaction ID
4-O-beta-D-mannopyranosyl-N-acetyl-D-glucosamine + phosphate = N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate sequential Bi Bi mechanism Bacteroides thetaiotaomicron

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
30
-
pH 5.5, 30°C, substrate: N,N'-diacetylchitobiose Bacteroides thetaiotaomicron
258
-
pH 5.5, 30°C, substrate: N-acetyl-D-glucosamine Bacteroides thetaiotaomicron

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-O-beta-D-mannopyranosyl-N-acetyl-D-glucosamine + phosphate the enzyme is involved in the degradation of host-derived N-glycans Bacteroides thetaiotaomicron N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
r
4-O-beta-D-mannopyranosyl-N-acetyl-D-glucosamine + phosphate the enzyme is involved in the degradation of host-derived N-glycans Bacteroides thetaiotaomicron VPI-5482 N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
r
4-O-beta-D-mannosyl-N-acetyl-D-glucosamine + phosphate
-
Bacteroides thetaiotaomicron N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
r
4-O-beta-D-mannosyl-N-acetyl-D-glucosamine + phosphate
-
Bacteroides thetaiotaomicron VPI-5482 N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
r
4-O-beta-N-acetyl-D-glucosaminyl-N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
Bacteroides thetaiotaomicron beta-N-acetyl-D-glucosaminyl-(1->4)-beta-N-acetyl-D-glucosaminyl-(1->4)-D-mannose + phosphate
-
r
4-O-beta-N-acetyl-D-glucosaminyl-N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
Bacteroides thetaiotaomicron VPI-5482 beta-N-acetyl-D-glucosaminyl-(1->4)-beta-N-acetyl-D-glucosaminyl-(1->4)-D-mannose + phosphate
-
r
beta-N-acetyl-D-glucosaminyl-(1->4)-beta-N-acetyl-D-glucosaminyl-(1->4)-D-mannose + phosphate
-
Bacteroides thetaiotaomicron 4-O-beta-N-acetyl-D-glucosaminyl-N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
r
beta-N-acetyl-D-glucosaminyl-(1->4)-beta-N-acetyl-D-glucosaminyl-(1->4)-D-mannose + phosphate
-
Bacteroides thetaiotaomicron VPI-5482 4-O-beta-N-acetyl-D-glucosaminyl-N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
r
additional information very low activity with D-glucose or D-mannose as acceptor and alpha-D-mannose 1-phosphate as donor Bacteroides thetaiotaomicron ?
-
?
additional information very low activity with D-glucose or D-mannose as acceptor and alpha-D-mannose 1-phosphate as donor Bacteroides thetaiotaomicron VPI-5482 ?
-
?
N,N'-diacetylchitobiose + alpha-D-mannose 1-phosphate
-
Bacteroides thetaiotaomicron beta-1,4-D-mannopyranosyl-beta-1,4-N-acetyl-D-glucosaminyl-beta-1,4-N-acetyl-D-glucosamine + phosphate
-
r
N,N'-diacetylchitobiose + alpha-D-mannose 1-phosphate
-
Bacteroides thetaiotaomicron VPI-5482 beta-1,4-D-mannopyranosyl-beta-1,4-N-acetyl-D-glucosaminyl-beta-1,4-N-acetyl-D-glucosamine + phosphate
-
r
N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
Bacteroides thetaiotaomicron 4-O-beta-D-mannosyl-N-acetyl-D-glucosamine + phosphate
-
r
N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
Bacteroides thetaiotaomicron VPI-5482 4-O-beta-D-mannosyl-N-acetyl-D-glucosamine + phosphate
-
r
N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
Bacteroides thetaiotaomicron 4-O-beta-D-mannopyranosyl-N-acetyl-D-glucosamine + phosphate
-
r
N-acetyl-D-glucosamine + alpha-D-mannose 1-phosphate
-
Bacteroides thetaiotaomicron VPI-5482 4-O-beta-D-mannopyranosyl-N-acetyl-D-glucosamine + phosphate
-
r

Subunits

Subunits Comment Organism
homotetramer 4 * 35000, SDS-PAGE Bacteroides thetaiotaomicron
homotetramer 4 * 35000, SDS-PAGE, 4 * 37823, amino acid sequence Bacteroides thetaiotaomicron
homotetramer 4 * 37823, calculated from sequence Bacteroides thetaiotaomicron

Synonyms

Synonyms Comment Organism
beta-1,4-D-mannosyl-N-acetyl-D-glucosamine phosphorylase
-
Bacteroides thetaiotaomicron
BT1033
-
Bacteroides thetaiotaomicron

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Bacteroides thetaiotaomicron

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
55
-
15 min, stable up to Bacteroides thetaiotaomicron
55
-
15 min stable up to Bacteroides thetaiotaomicron

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
22
-
4-O-beta-N-acetyl-D-glucosaminyl-N-acetyl-D-glucosamine pH 5.5, 30°C Bacteroides thetaiotaomicron
22
-
N,N'-diacetylchitobiose pH 5.5, 30°C, synthetic reaction Bacteroides thetaiotaomicron
25
-
alpha-D-mannose 1-phosphate pH 5.5, 30°C Bacteroides thetaiotaomicron
25
-
alpha-D-mannose 1-phosphate pH 5.5, 30°C, cosubstrate: N-acetyl-D-glucosamine, synthetic reaction Bacteroides thetaiotaomicron
37
-
N-acetyl-D-glucosamine pH 5.5, 30°C Bacteroides thetaiotaomicron
37
-
N-acetyl-D-glucosamine pH 5.5, 30°C, synthetic reaction Bacteroides thetaiotaomicron
240
-
phosphate pH 5.5, 30°C, cosubstrate: 4-O-beta-D-mannopyranosyl-N-acetyl-D-glucosamine, phosphorolytic reaction Bacteroides thetaiotaomicron
240
-
4-O-beta-D-mannopyranosyl-N-acetyl-D-glucosamine pH 5.5, 30°C, cosubstrate: phosphate, phosphorolytic reaction Bacteroides thetaiotaomicron

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5
-
-
Bacteroides thetaiotaomicron
5.5
-
phosphorolytic reaction Bacteroides thetaiotaomicron
5.5
-
synthetic reaction Bacteroides thetaiotaomicron

pH Stability

pH Stability pH Stability Maximum Comment Organism
4.5 10.5 4°C, 24 h, stable Bacteroides thetaiotaomicron
4.5 10.5 4°C, 24 h stable Bacteroides thetaiotaomicron

General Information

General Information Comment Organism
physiological function the enzyme is involved in the degradation of host-derived N-glycans Bacteroides thetaiotaomicron

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.8
-
N,N'-diacetylchitobiose pH 5.5, 30°C, synthetic reaction Bacteroides thetaiotaomicron
1.9
-
beta-1,4-D-mannopyranosyl-beta-1,4-N-acetyl-D-glucosaminyl-beta-1,4-N-acetyl-D-glucosamine pH 5.5, 30°C, cosubstrate: phosphate, phosphorolytic reaction Bacteroides thetaiotaomicron
9.6
-
alpha-D-mannose 1-phosphate pH 5.5, 30°C, cosubstrate: N-acetyl-D-glucosamine, synthetic reaction Bacteroides thetaiotaomicron
9.6
-
N-acetyl-D-glucosamine pH 5.5, 30°C, synthetic reaction Bacteroides thetaiotaomicron
110
-
4-O-beta-D-mannopyranosyl-N-acetyl-D-glucosamine pH 5.5, 30°C, cosubstrate: phosphate, phosphorolytic reaction Bacteroides thetaiotaomicron