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Literature summary for 3.2.1.24 extracted from

  • Hansen, D.K.; Webb, H.; Nielsen, J.W.; Harris, P.; Winther, J.R.; Willemoes, M.
    Mutational analysis of divalent metal ion binding in the active site of class II alpha-mannosidase from Sulfolobus solfataricus (2015), Biochemistry, 24, 2032-2039.
    View publication on PubMed
Search for more literature for 3.2.1.24

Protein Variants

Protein Variants Comment Organism
D534A mutant enzyme shows no detectable activity (kcat below 0.01 s-1) within the pH range of 3-7 and with Co2+ as the activating divalent metal ion Saccharolobus solfataricus
D534Q mutant enzyme shows no detectable activity (kcat below 0.01 s-1) within the pH range of 3-7 and with Co2+ as the activating divalent metal ion Saccharolobus solfataricus
H228E/H533E mutant enzyme shows no detectable activity (kcat below 0.01 s-1) within the pH range of 3-7 and with Co2+ as the activating divalent metal ion Saccharolobus solfataricus
H228Q the mutant enzyme is inhibited at increasing Zn2+ concentrations. The catalytic rate is reduced for all enzymes compared to that of the wild-type enzyme, although less dramatically with some activating metal ions. The mutant enzyme has a structural integrity in terms of thermostability similar to that of the wild-type enzyme Saccharolobus solfataricus
H228Q/H533Q mutant enzyme shows no detectable activity (kcat below 0.01 s-1) within the pH range of 3-7 and with Co2+ as the activating divalent metal ion Saccharolobus solfataricus
H533E the mutant enzyme is inhibited at increasing Zn2+ concentrations. The catalytic rate is reduced for all enzymes compared to that of the wild-type enzyme, although less dramatically with some activating metal ions The mutant enzyme has a structural integrity in terms of thermostability similar to that of the wild-type enzyme Saccharolobus solfataricus
H533Q the mutant enzyme is inhibited at increasing Zn2+ concentrations. The catalytic rate is reduced for all enzymes compared to that of the wild-type enzyme, although less dramatically with some activating metal ions The mutant enzyme has a structural integrity in terms of thermostability similar to that of the wild-type enzyme Saccharolobus solfataricus

Inhibitors

Inhibitors Comment Organism Structure
Zn2+ mutant enzymes H228Q, H533E, and H533Q, are all inhibited by high Zn2+ concentrations (1 mM) Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.13
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Co2+, mutant enzyme H533Q Saccharolobus solfataricus
0.35
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Co2+, wild-type enzyme Saccharolobus solfataricus
0.47
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Mn2+, wild-type enzyme Saccharolobus solfataricus
0.7
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Mn2+, mutant enzyme H533Q Saccharolobus solfataricus
0.74
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Zn2+, wild-type enzyme Saccharolobus solfataricus
1.1
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 0.1 mM Zn2+, mutant enzyme H533Q Saccharolobus solfataricus
1.1
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Cd2+, mutant enzyme H533Q Saccharolobus solfataricus
1.1
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Zn2+, mutant enzyme H228E Saccharolobus solfataricus
1.4
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Co2+, mutant enzyme H228E Saccharolobus solfataricus
1.7
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Co2+, mutant enzyme H228Q Saccharolobus solfataricus
1.9
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Cd2+, wild-type enzyme Saccharolobus solfataricus
1.9
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Zn2+, mutant enzyme H533Q Saccharolobus solfataricus
2.5
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Zn2+, mutant enzyme H228Q Saccharolobus solfataricus
2.7
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Cd2+, mutant enzyme H228E Saccharolobus solfataricus
2.9
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Co2+, mutant enzyme H533E Saccharolobus solfataricus

Metals/Ions

Metals/Ions Comment Organism Structure
Cd2+ in the wild-type enzyme Cd2+ stimulates hydrolysis of 4-nitrophenyl alpha-D-mannoside to a 4fold lesser extent than Co2+. The side chains at positions 228 and 533 are involved in binding the activating metal ion as their primary function Saccharolobus solfataricus
Co2+ Co2+ is by far the most efficient metal ion in stimulating hydrolysis of 4-nitrophenyl alpha-D-mannoside. The side chains at positions 228 and 533 are involved in binding the activating metal ion as their primary function Saccharolobus solfataricus
Mn2+ in the wild-type enzyme Mn2+ stimulates hydrolysis of 4-nitrophenyl alpha-D-mannoside to a 3.2fold lesser extent than Co2+. The side chains at positions 228 and 533 are involved in binding the activating metal ion as their primary function Saccharolobus solfataricus
Zn2+ in the wild-type enzyme Zn2+ stimulates hydrolysis of 4-nitrophenyl alpha-D-mannoside to a 3.6fold lesser extent than Co2+. The side chains at positions 228 and 533 are involved in binding the activating metal ion as their primary function Saccharolobus solfataricus

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus Q97UK5
-
-
Saccharolobus solfataricus P2 Q97UK5
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Saccharolobus solfataricus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl alpha-D-mannoside + H2O
-
 Saccharolobus solfataricus 4-nitrophenol + alpha-D-mannose
-
 ?
4-nitrophenyl alpha-D-mannoside + H2O
-
 Saccharolobus solfataricus P2 4-nitrophenol + alpha-D-mannose
-
 ?

Synonyms

Synonyms Comment Organism
class II alpha-mannosidase
-
 Saccharolobus solfataricus
ManA
-
 Saccharolobus solfataricus
SSO3006 locus name Saccharolobus solfataricus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
 assay at Saccharolobus solfataricus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30
-
 inactive below Saccharolobus solfataricus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.088
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Mn2+, mutant enzyme H533Q Saccharolobus solfataricus
0.099
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Co2+, mutant enzyme H533Q Saccharolobus solfataricus
0.118
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Zn2+, mutant enzyme H228E Saccharolobus solfataricus
0.4
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Cd2+, mutant enzyme H533Q Saccharolobus solfataricus
0.6
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Zn2+, mutant enzyme H228Q Saccharolobus solfataricus
0.8
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Co2+, mutant enzyme H228E Saccharolobus solfataricus
1.18
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Cd2+, mutant enzyme H228E Saccharolobus solfataricus
1.42
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Zn2+, mutant enzyme H533Q Saccharolobus solfataricus
1.6
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 0.1 mM Zn2+, mutant enzyme H533Q Saccharolobus solfataricus
1.6
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Co2+, mutant enzyme H228Q Saccharolobus solfataricus
4.3
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Co2+, mutant enzyme H533E Saccharolobus solfataricus
4.8
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Mn2+, wild-type enzyme Saccharolobus solfataricus
6.6
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Zn2+, wild-type enzyme Saccharolobus solfataricus
11.3
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Co2+, wild-type enzyme Saccharolobus solfataricus
15.3
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Cd2+, wild-type enzyme Saccharolobus solfataricus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5
-
 wild-type enzyme and mutant enzymes H228E, H228Q, H533E, and H533Q Saccharolobus solfataricus

pH Range

pH Minimum pH Maximum Comment Organism
4.2 6 pH 4.2: about 35% of maximal activity, pH 6.0: about 40% of maximal activity, wild-type enzyme Saccharolobus solfataricus

pH Stability

pH Stability pH Stability Maximum Comment Organism
4.5
-
 instability below pH 4.5 Saccharolobus solfataricus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.014
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Mn2+, mutant enzyme H533E. KM is too high (above 10 mM for 4-nitrophenyl alpha-D-mannoside or.0.1 mM for Mn2+) to allow for determination of kinetic parameters other than kcat/KM Saccharolobus solfataricus
0.019
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Mn2+, mutant enzyme H228E. KM is too high (above 10 mM for 4-nitrophenyl alpha-D-mannoside or.0.1 mM for Mn2+) to allow for determination of kinetic parameters other than kcat/KM Saccharolobus solfataricus
0.038
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Mn2+, mutant enzyme H228Q. KM is too high (above 10 mM for 4-nitrophenyl alpha-D-mannoside or.0.1 mM for Mn2+) to allow for determination of kinetic parameters other than kcat/KM Saccharolobus solfataricus
0.046
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Cd2+, mutant enzyme H533E. KM is too high (above 10 mM for 4-nitrophenyl alpha-D-mannoside or.0.1 mM for Mn2+) to allow for determination of kinetic parameters other than kcat/KM Saccharolobus solfataricus
0.0732
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Zn2+, mutant enzyme H533E. KM is too high (above 10 mM for 4-nitrophenyl alpha-D-mannoside or.0.1 mM for Mn2+) to allow for determination of kinetic parameters other than kcat/KM Saccharolobus solfataricus
0.11
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Zn2+, mutant enzyme H228E Saccharolobus solfataricus
0.13
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Mn2+, mutant enzyme H533Q Saccharolobus solfataricus
0.24
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 0.1 mM Zn2+, mutant enzyme H533E. KM is too high (above 10 mM for 4-nitrophenyl alpha-D-mannoside or.0.1 mM for Mn2+) to allow for determination of kinetic parameters other than kcat/KM Saccharolobus solfataricus
0.24
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Zn2+, mutant enzyme H228Q Saccharolobus solfataricus
0.36
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Cd2+, mutant enzyme H533Q Saccharolobus solfataricus
0.43
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Cd2+, mutant enzyme H228E Saccharolobus solfataricus
0.53
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Cd2+, mutant enzyme H228Q. KM is too high (above 10 mM for 4-nitrophenyl alpha-D-mannoside or.0.1 mM for Mn2+) to allow for determination of kinetic parameters other than kcat/KM Saccharolobus solfataricus
0.57
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Co2+, mutant enzyme H228E Saccharolobus solfataricus
0.75
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Zn2+, mutant enzyme H533Q Saccharolobus solfataricus
0.76
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Co2+, mutant enzyme H533Q Saccharolobus solfataricus
0.94
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Co2+, mutant enzyme H228Q Saccharolobus solfataricus
1.5
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 0.1 mM Zn2+, mutant enzyme H533Q Saccharolobus solfataricus
1.5
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Co2+, mutant enzyme H533E Saccharolobus solfataricus
8.1
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Cd2+, wild-type enzyme Saccharolobus solfataricus
8.9
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Zn2+, wild-type enzyme Saccharolobus solfataricus
10.2
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Mn2+, wild-type enzyme Saccharolobus solfataricus
32.3
-
 4-nitrophenyl alpha-D-mannoside pH 5.0, 70°C, 1 mM Co2+, wild-type enzyme Saccharolobus solfataricus