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Literature summary for 2.4.1.19 extracted from
- Mutations of lysine 47 in cyclodextrin glycosyltransferase from Paenibacillus macerans enhance beta-cyclodextrin specificity (2009), J. Agric. Food Chem., 57, 8386-8391.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| subcloning in Escherichia coli strain JM109, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Paenibacillus macerans |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| K47H | site-directed mutagenesis, the mutant shows a shift in product specificity, slight enhancement of beta-cyclodextrin production and slight reduction of alpha-cyclodextrin production compared to the wild-type enzyme, the mutant enzyme exhibits lower stability compared to the wild-type enzyme | Paenibacillus macerans |
| K47L | site-directed mutagenesis, the mutant shows a shift in product specificity, enhancement of beta-cyclodextrin production and reduction of alpha-cyclodextrin production compared to the wild-type enzyme, the mutant enzyme exhibits lower stability compared to the wild-type enzyme | Paenibacillus macerans |
| K47R | site-directed mutagenesis, the mutant shows a shift in product specificity, slight enhancement of beta-cyclodextrin production and slight reduction of alpha-cyclodextrin production compared to the wild-type enzyme, the mutant enzyme exhibits lower stability compared to the wild-type enzyme | Paenibacillus macerans |
| K47S | site-directed mutagenesis, the mutant shows a shift in product specificity, enhancement of beta-cyclodextrin production and reduction of alpha-cyclodextrin production compared to the wild-type enzyme, the mutant enzyme exhibits lower stability compared to the wild-type enzyme | Paenibacillus macerans |
| K47T | site-directed mutagenesis, the mutant shows a shift in product specificity, enhancement of beta-cyclodextrin production and reduction of alpha-cyclodextrin production compared to the wild-type enzyme, the mutant enzyme exhibits lower stability compared to the wild-type enzyme | Paenibacillus macerans |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Paenibacillus macerans | - |
CCTCCM 203062 | - |
| Paenibacillus macerans JFB05-01 | - |
CCTCCM 203062 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Paenibacillus macerans |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 14.4 | - |
beta-cyclodextrin-forming activity, recombinant wild-type enzyme | Paenibacillus macerans |
| 21.7 | - |
beta-cyclodextrin-forming activity, recombinant mutant K47R | Paenibacillus macerans |
| 35.9 | - |
beta-cyclodextrin-forming activity, recombinant mutant K47H | Paenibacillus macerans |
| 45.9 | - |
alpha-cyclodextrin-forming activity, recombinant mutant K47L | Paenibacillus macerans |
| 46.4 | - |
alpha-cyclodextrin-forming activity, recombinant mutant K47S | Paenibacillus macerans |
| 47.4 | - |
alpha-cyclodextrin-forming activity, recombinant mutant K47T | Paenibacillus macerans |
| 51.4 | - |
beta-cyclodextrin-forming activity, recombinant mutant K47T | Paenibacillus macerans |
| 51.5 | - |
beta-cyclodextrin-forming activity, recombinant mutant K47S | Paenibacillus macerans |
| 53.1 | - |
beta-cyclodextrin-forming activity, recombinant mutant K47L | Paenibacillus macerans |
| 62.9 | - |
alpha-cyclodextrin-forming activity, recombinant mutant K47H | Paenibacillus macerans |
| 77.3 | - |
alpha-cyclodextrin-forming activity, recombinant mutant K47R | Paenibacillus macerans |
| 85.1 | - |
alpha-cyclodextrin-forming activity, recombinant wild-type enzyme | Paenibacillus macerans |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | the enzyme performs formation of alpha-, beta- and gamma-cyclodextrin. Lys47 is important for the alpha-cyclization reaction. Enhancement of beta-cyclodextrin specificity might be due to weakening or removal of hydrogen-bonding interactions between the side chain of residue 47 and the bent intermediate specific for alpha-cyclodextrin formation | Paenibacillus macerans | ? | - |
? |  |
| additional information | the enzyme performs formation of alpha-, beta- and gamma-cyclodextrin. Lys47 is important for the alpha-cyclization reaction. Enhancement of beta-cyclodextrin specificity might be due to weakening or removal of hydrogen-bonding interactions between the side chain of residue 47 and the bent intermediate specific for alpha-cyclodextrin formation | Paenibacillus macerans JFB05-01 | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| CGTase | - |
Paenibacillus macerans |
| cyclodextrin glycosyltransferase | - |
Paenibacillus macerans |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | - |
assay at | Paenibacillus macerans |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | - |
the recombinant wild-type enzyme has a half-life of 8.0 h, while the recombinant K47 mutants show half-lives of 4.8-7.2 h | Paenibacillus macerans |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6 | - |
assay at | Paenibacillus macerans |
General Information
| General Information | Comment | Organism |
|---|---|---|
| physiological function | the enzyme is responsible for cyclodextrin formation. Cyclodextrins are cyclic, nonreducing oligo-glucopyranose molecules linked via alpha(1,4)-glycosidic bonds mainly consisting of six, seven, or eight glucose residue, alpha-, beta-, or gamma-cyclodextrin, respectively | Paenibacillus macerans |
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