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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Paenibacillus macerans | - |
CCTCCM 203062 | - |
Paenibacillus macerans JFB05-01 | - |
CCTCCM 203062 | - |
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 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 | 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 | Comment | Organism |
---|---|---|
CGTase | - |
Paenibacillus macerans |
cyclodextrin glycosyltransferase | - |
Paenibacillus macerans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
assay at | Paenibacillus macerans |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
assay at | Paenibacillus macerans |
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 |