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Literature summary for 2.4.1.19 extracted from
- Site-saturation engineering of lysine 47 in cyclodextrin glycosyltransferase from Paenibacillus macerans to enhance substrate specificity towards maltodextrin for enzymatic synthesis of 2-O-D-glucopyranosyl-L-ascorbic acid (AA-2G) (2013), Appl. Microbiol. Biotechnol., 97, 5851-5860.
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| K47F | improved synthesis of 2-O-D-glucopyranosyl-L-ascorbic acid with maltodextrin as glucosyl donor, 30% increase in yield. Mutation leads to relatively lower cyclization activities and higher disproportionation activities. The enhancement of maltodextrin specificity may be due to the short residue chain and the removal of hydrogen bonding interactions between the side chain of residue 47 and the sugar at -3 subsite | Paenibacillus macerans |
| K47L | improved synthesis of 2-O-D-glucopyranosyl-L-ascorbic acid with maltodextrin as glucosyl donor, highest titer of product among the mutants tested, 57% increase in yield. Mutation leads to relatively lower cyclization activities and higher disproportionation activities. The enhancement of maltodextrin specificity may be due to the short residue chain and the removal of hydrogen bonding interactions between the side chain of residue 47 and the sugar at -3 subsite | Paenibacillus macerans |
| K47V | improved synthesis of 2-O-D-glucopyranosyl-L-ascorbic acid with maltodextrin as glucosyl donor, 48% increase in yield. Mutation leads to relatively lower cyclization activities and higher disproportionation activities. The enhancement of maltodextrin specificity may be due to the short residue chain and the removal of hydrogen bonding interactions between the side chain of residue 47 and the sugar at -3 subsite | Paenibacillus macerans |
| K47W | improved synthesis of 2-O-D-glucopyranosyl-L-ascorbic acid with maltodextrin as glucosyl donor, 24% increase in yield. Mutation leads to relatively lower cyclization activities and higher disproportionation activities. The enhancement of maltodextrin specificity may be due to the short residue chain and the removal of hydrogen bonding interactions between the side chain of residue 47 and the sugar at -3 subsite | Paenibacillus macerans |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.48 | - |
maltodextrin | mutant K47V, pH 5.5, 37°C, average molecular weight of 2150.68 Da for maltodextrin | Paenibacillus macerans |  |
| 0.49 | - |
maltodextrin | mutant K47L, pH 5.5, 37°C, average molecular weight of 2150.68 Da for maltodextrin | Paenibacillus macerans | |
| 0.52 | - |
maltodextrin | mutant K47F, pH 5.5, 37°C, average molecular weight of 2150.68 Da for maltodextrin | Paenibacillus macerans | |
| 0.55 | - |
maltodextrin | mutant K47W, pH 5.5, 37°C, average molecular weight of 2150.68 Da for maltodextrin | Paenibacillus macerans | |
| 0.64 | - |
maltodextrin | wild-type, pH 5.5, 37°C, average molecular weight of 2150.68 Da for maltodextrin | Paenibacillus macerans | |
| 38.3 | - |
L-ascorbic acid | wild-type, pH 5.5, 37°C, average molecular weight of 2150.68 Da for maltodextrin | Paenibacillus macerans | |
| 44.7 | - |
L-ascorbic acid | mutant K47W, pH 5.5, 37°C, average molecular weight of 2150.68 Da for maltodextrin | Paenibacillus macerans | |
| 46.1 | - |
L-ascorbic acid | mutant K47F, pH 5.5, 37°C, average molecular weight of 2150.68 Da for maltodextrin | Paenibacillus macerans | |
| 46.9 | - |
L-ascorbic acid | mutant K47V, pH 5.5, 37°C, average molecular weight of 2150.68 Da for maltodextrin | Paenibacillus macerans | |
| 51.7 | - |
L-ascorbic acid | mutant K47L, pH 5.5, 37°C, average molecular weight of 2150.68 Da for maltodextrin | Paenibacillus macerans | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 75000 | - |
x * 75000, SDS-PAGE | Paenibacillus macerans |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Paenibacillus macerans | O52766 | - |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| L-ascorbic acid + beta-cyclodextrin | - |
Paenibacillus macerans | L-ascorbic acid-2-O-alpha-D-glucoside + ? | - |
? | |
| L-ascorbic acid + maltodextrin | - |
Paenibacillus macerans | L-ascorbic acid-2-O-alpha-D-glucoside + ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 75000, SDS-PAGE | Paenibacillus macerans |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| CGT | - |
Paenibacillus macerans |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 36 | - |
optimum both for wild-type and mutant strains | Paenibacillus macerans |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 28 | 44 | - |
Paenibacillus macerans |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.004 | - |
maltodextrin | mutant K47W, pH 5.5, 37°C, average molecular weight of 2150.68 Da for maltodextrin | Paenibacillus macerans | |
| 0.004 | - |
maltodextrin | wild-type, pH 5.5, 37°C, average molecular weight of 2150.68 Da for maltodextrin | Paenibacillus macerans | |
| 0.005 | - |
maltodextrin | mutant K47F, pH 5.5, 37°C, average molecular weight of 2150.68 Da for maltodextrin | Paenibacillus macerans | |
| 0.005 | - |
maltodextrin | mutant K47V, pH 5.5, 37°C, average molecular weight of 2150.68 Da for maltodextrin | Paenibacillus macerans | |
| 0.006 | - |
maltodextrin | mutant K47L, pH 5.5, 37°C, average molecular weight of 2150.68 Da for maltodextrin | Paenibacillus macerans | |
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