Literature summary for 2.4.1.339 extracted from

Tsuda, T.; Nihira, T.; Chiku, K.; Suzuki, E.; Arakawa, T.; Nishimoto, M.; Kitaoka, M.; Nakai, H.; Fushinobu, S.
Characterization and crystal structure determination of beta-1,2-mannobiose phosphorylase from Listeria innocua (2015), FEBS Lett., 589, 3816-3821.

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Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli as a C-terminally (His)6-tagged protein	Listeria innocua

Crystallization (Commentary)

Crystallization (Comment)	Organism
sitting drop vapor diffusion method at 20°C, crystals complexed with beta-1,2-Man2 and beta-1,2-Man3 are prepared by soaking in the cryoprotectant solution supplemented with 50 mM beta-1,2-Man2 or 10 mM beta-1,2-Man3. beta-1,2-Mannotriose is bound in a U-shape, interacting with a phosphate analog at both ends. Lin0857 has a unique dimer structure connected by a loop, and a significant openclose loop displacement iss observed for substrate entry. A long loop, which is exclusively present in Lin0857, covers the active site to limit the pocket size. A structural basis for substrate recognition and phosphorolysis is provided	Listeria innocua

Crystallization (Comment)

Organism

sitting drop vapor diffusion method at 20°C, crystals complexed with beta-1,2-Man2 and beta-1,2-Man3 are prepared by soaking in the cryoprotectant solution supplemented with 50 mM beta-1,2-Man2 or 10 mM beta-1,2-Man3. beta-1,2-Mannotriose is bound in a U-shape, interacting with a phosphate analog at both ends. Lin0857 has a unique dimer structure connected by a loop, and a significant openclose loop displacement iss observed for substrate entry. A long loop, which is exclusively present in Lin0857, covers the active site to limit the pocket size. A structural basis for substrate recognition and phosphorolysis is provided

Listeria innocua

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.13	-	phosphate	30°C, pH 7, cosubstrate: beta-D-mannopyranosyl-(1->2)-beta-D-mannopyranosyl-(1->2)-D-mannopyranose	Listeria innocua
0.19	-	phosphate	30°C, pH 7, cosubstrate: beta-D-mannopyranosyl-(1->2)-D-mannopyranose	Listeria innocua
0.44	-	beta-D-mannopyranosyl-(1->2)-D-mannopyranose	30°C, pH 7	Listeria innocua
1.5	-	D-fructose	30°C, pH 6	Listeria innocua
2.1	-	D-mannopyranose	30°C, pH 6	Listeria innocua
3.1	-	alpha-D-mannose 1-phosphate	30°C, pH 6	Listeria innocua
4.1	-	beta-D-mannopyranosyl-(1->2)-beta-D-mannopyranosyl-(1->2)-D-mannopyranose	30°C, pH 7	Listeria innocua
5.1	-	beta-D-mannopyranosyl-(1->2)-D-mannopyranose	30°C, pH 6	Listeria innocua

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
40000	-	2 * 40000, SDS-PAGE	Listeria innocua
88000	-	gel filtration	Listeria innocua

Organism

Organism	UniProt	Comment	Textmining
Listeria innocua	Q92DF6	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Listeria innocua

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
beta-D-mannopyranosyl-(1->2)-beta-D-mannopyranosyl-(1->2)-D-mannopyranose + phosphate	the enzyme preferentially phosphorolyzes beta-1,2-Man2 over beta-1,2-Man3	Listeria innocua	beta-D-mannopyranosyl-(1->2)-D-mannopyranose + alpha-D-mannose 1-phosphate	-	r
beta-D-mannopyranosyl-(1->2)-D-mannopyranose + alpha-D-mannose 1-phosphate	no production of tetrasaccharide is detected	Listeria innocua	beta-D-mannopyranosyl-(1->2)-beta-D-mannopyranosyl-(1->2)-D-mannopyranose + phosphate	-	r
beta-D-mannopyranosyl-(1->2)-D-mannopyranose + phosphate	the enzyme preferentially phosphorolyzes beta-1,2-Man2 over beta-1,2-Man3	Listeria innocua	D-mannopyranose + alpha-D-mannose 1-phosphate	-	r
D-fructose + alpha-D-mannose 1-phosphate	-	Listeria innocua	? + phosphate	-	r
D-mannopyranose + alpha-D-mannose 1-phosphate	-	Listeria innocua	beta-D-mannopyranosyl-(1->2)-D-mannopyranose + phosphate	-	r

Subunits

Subunits	Comment	Organism
dimer	2 * 40000, SDS-PAGE	Listeria innocua

Synonyms

Synonyms	Comment	Organism
Lin0857	-	Listeria innocua

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
50	-	stable up to 50°C during 15 min of incubation	Listeria innocua

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
3.9	-	D-fructose	30°C, pH 6	Listeria innocua
12	-	beta-D-mannopyranosyl-(1->2)-D-mannopyranose	30°C, pH 6	Listeria innocua
12	-	beta-D-mannopyranosyl-(1->2)-beta-D-mannopyranosyl-(1->2)-D-mannopyranose	30°C, pH 7	Listeria innocua
31	-	D-mannopyranose	30°C, pH 6	Listeria innocua
33	-	alpha-D-mannose 1-phosphate	30°C, pH 6	Listeria innocua
35	-	beta-D-mannopyranosyl-(1->2)-D-mannopyranose	30°C, pH 7	Listeria innocua

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6	-	synthetic reaction	Listeria innocua
7	-	phosphorolytic reaction	Listeria innocua

pH Range

pH Minimum	pH Maximum	Comment	Organism
4.5	7.5	pH 4.5: about 60% of maximal activity, pH 7.5: about 60% of maximal activity, synthetic reaction	Listeria innocua
5.5	8	pH 5.5: about 75% of maximal activity, pH 8.0: about 60% of maximal activity, phosphorolytic reaction	Listeria innocua

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
5.5	9	4°C, 24 h, stable	Listeria innocua

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
2.3	-	beta-D-mannopyranosyl-(1->2)-D-mannopyranose	30°C, pH 6	Listeria innocua
2.7	-	D-fructose	30°C, pH 6	Listeria innocua
3	-	beta-D-mannopyranosyl-(1->2)-beta-D-mannopyranosyl-(1->2)-D-mannopyranose	30°C, pH 7	Listeria innocua
11	-	alpha-D-mannose 1-phosphate	30°C, pH 6	Listeria innocua
15	-	D-mannopyranose	30°C, pH 6	Listeria innocua
79	-	beta-D-mannopyranosyl-(1->2)-D-mannopyranose	30°C, pH 7	Listeria innocua