Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain JM109 | Neisseria polysaccharea |
Protein Variants | Comment | Organism |
---|---|---|
E328Q | site-directed mutagenesis, inactive mutant | Neisseria polysaccharea |
R226N | site-directed mutagenesis, compared to the wild-type enzyme, the mutant shows a 10fold enhancement in the catalytic efficiency and a nearly twofold higher production of an insoluble amylose-like polymer | Neisseria polysaccharea |
R226X | site-directed mutagenesis, the single site mutants, except R226N, show reduced activity compare to the wild-type enzyme | Neisseria polysaccharea |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0123 | - |
maltotriose | pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea | |
0.014 | - |
maltobiose | pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea | |
0.024 | - |
maltopentaose | pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea | |
0.026 | - |
maltotetraose | pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea | |
11 | - |
maltobiose | pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea | |
16 | - |
maltotriose | pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea | |
30 | - |
maltotetraose | pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea | |
61 | - |
maltopentaose | pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Neisseria polysaccharea | amylosucrase from Neisseria polysaccharea is a transglucosylase that synthesizes an insoluble amylose-like polymer from sole substrate sucrose, product isolation and analysis, overview | ? | - |
? | |
sucrose + maltobiose | Neisseria polysaccharea | - |
D-fructose + maltotriose | - |
? | |
sucrose + maltopentaose | Neisseria polysaccharea | - |
D-fructose + maltohexaose | - |
? | |
sucrose + maltotetraose | Neisseria polysaccharea | - |
D-fructose + maltopentaose | - |
? | |
sucrose + maltotriose | Neisseria polysaccharea | - |
D-fructose + maltotetraose | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Neisseria polysaccharea | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | amylosucrase from Neisseria polysaccharea is a transglucosylase that synthesizes an insoluble amylose-like polymer from sole substrate sucrose, product isolation and analysis, overview | Neisseria polysaccharea | ? | - |
? | |
sucrose + maltobiose | - |
Neisseria polysaccharea | D-fructose + maltotriose | - |
? | |
sucrose + maltopentaose | - |
Neisseria polysaccharea | D-fructose + maltohexaose | - |
? | |
sucrose + maltotetraose | - |
Neisseria polysaccharea | D-fructose + maltopentaose | - |
? | |
sucrose + maltotriose | - |
Neisseria polysaccharea | D-fructose + maltotetraose | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | amylosucrases adopt a deep pocket topology of about 15 A with the catalytic triad located at the bottom | Neisseria polysaccharea |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Neisseria polysaccharea |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.1 | - |
maltobiose | pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea | |
3.1 | - |
sucrose | with maltobiose, pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea | |
4.8 | - |
maltotriose | pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea | |
4.8 | - |
sucrose | with maltotriose, pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea | |
7.5 | - |
maltotetraose | pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea | |
7.5 | - |
sucrose | with maltotetraose, pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea | |
14 | - |
maltobiose | pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea | |
14 | - |
sucrose | with maltobiose, pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea | |
33 | - |
maltotriose | pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea | |
33 | - |
sucrose | with maltotriose, pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea | |
47 | - |
maltopentaose | pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea | |
47 | - |
sucrose | with maltopentaose, pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea | |
67 | - |
maltotetraose | pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea | |
67 | - |
sucrose | with maltotetraose, pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea | |
173 | - |
maltopentaose | pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea | |
173 | - |
sucrose | with maltopentaose, pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Neisseria polysaccharea |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the GH13 family. Amylosucrases adopt a deep pocket topology of about 15 A with the catalytic triad located at the bottom | Neisseria polysaccharea |
additional information | active site topology and structure coparisons, overview | Neisseria polysaccharea |
additional information | Arg226, was proposed from molecular modeling studies to play an important role in the formation of the active site topology and in the accessibility of ligands to the catalytic site | Neisseria polysaccharea |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.25 | - |
maltotetraose | pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea | |
0.28 | - |
maltobiose | pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea | |
0.3 | - |
maltotriose | pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea | |
0.77 | - |
maltopentaose | pH 7.0, 30°C, recombinant wild-type enzyme | Neisseria polysaccharea | |
0.99 | - |
maltobiose | pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea | |
2.6 | - |
maltotetraose | pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea | |
2.7 | - |
maltotriose | pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea | |
7.2 | - |
maltopentaose | pH 7.0, 30°C, recombinant mutant R226N | Neisseria polysaccharea |