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Literature summary extracted from

  • Ye, Y.; Saburi, W.; Odaka, R.; Kato, K.; Sakurai, N.; Komoda, K.; Nishimoto, M.; Kitaoka, M.; Mori, H.; Yao, M.
    Structural insights into the difference in substrate recognition of two mannoside phosphorylases from two GH130 subfamilies (2016), FEBS Lett., 590, 828-837.
    View publication on PubMed

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.1.281 crystal structure with/without 4-O-beta-D-mannosyl-D-glucose. His245 of loop 3 forms a hydrogen-bond network with the substrate through a water molecule, and is indispensible for substrate binding Ruminococcus albus
2.4.1.319 in presence and absence of beta-(1->4)-mannobiose, and comparison with 4-O-beta-D-mannosyl-D-glucose phosphorylase MP1. The structures of the two enzymes differ at the +1 subsite of the substrate-binding pocket. Three loops are proposed to determine the different substrate specificities. Loop2 restricts the derivatives at the C6 position of the +1 substrate residue in beta-(1,4)-mannooligosaccharide phosphorylase MP2 Ruminococcus albus

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.281 H245A 1.47% of wild-type phosphorolytic activity Ruminococcus albus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.281 2.41
-
 alpha-D-mannose 1-phosphate wild-type, pH 6.5, 37°C Ruminococcus albus
2.4.1.281 25.9
-
 D-glucose wild-type, pH 6.5, 37°C Ruminococcus albus
2.4.1.281 36.5
-
 alpha-D-mannose 1-phosphate mutant H245A, pH 6.5, 37°C Ruminococcus albus
2.4.1.281 319
-
 D-glucose mutant H245A, pH 6.5, 37°C Ruminococcus albus

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.281 Ruminococcus albus E6UIS7
-
-
2.4.1.281 Ruminococcus albus DSM 20455 E6UIS7
-
-
2.4.1.319 Ruminococcus albus E6UBR9
-
-
2.4.1.319 Ruminococcus albus DSM 20455 E6UBR9
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.281 D-glucose + alpha-D-mannose 1-phosphate
-
 Ruminococcus albus 4-O-beta-D-mannopyranosyl-D-glucopyranose + phosphate
-
 ?
2.4.1.281 D-glucose + alpha-D-mannose 1-phosphate
-
 Ruminococcus albus DSM 20455 4-O-beta-D-mannopyranosyl-D-glucopyranose + phosphate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.1.281 MP1
-
 Ruminococcus albus
2.4.1.281 Rumal_0852
-
 Ruminococcus albus
2.4.1.319 MP2
-
 Ruminococcus albus
2.4.1.319 Rumal_0099
-
 Ruminococcus albus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.1.281 2.76
-
 alpha-D-mannose 1-phosphate mutant H245A, pH 6.5, 37°C Ruminococcus albus
2.4.1.281 19.3
-
 D-glucose mutant H245A, pH 6.5, 37°C Ruminococcus albus
2.4.1.281 52.1
-
 alpha-D-mannose 1-phosphate wild-type, pH 6.5, 37°C Ruminococcus albus
2.4.1.281 126
-
 D-glucose wild-type, pH 6.5, 37°C Ruminococcus albus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.4.1.281 0.061
-
 D-glucose mutant H245A, pH 6.5, 37°C Ruminococcus albus
2.4.1.281 0.076
-
 alpha-D-mannose 1-phosphate mutant H245A, pH 6.5, 37°C Ruminococcus albus
2.4.1.281 4.86
-
 D-glucose wild-type, pH 6.5, 37°C Ruminococcus albus
2.4.1.281 21.6
-
 alpha-D-mannose 1-phosphate wild-type, pH 6.5, 37°C Ruminococcus albus