Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Paenibacillus macerans |
Protein Variants | Comment | Organism |
---|---|---|
Q265K | the mutant shows 15% cyclization (alpha-cyclodextrin-forming) and 236% hydrolysis (starch-degrading) activities compared to the wild type enzyme | Paenibacillus macerans |
Q265K | the mutant with enhanced maltodextrin specificity produces higher 2-O-D-glucopyranosyl-L-ascorbic acid yields than the wild type enzyme. Compared with the wild type enzyme, the mutant has lower cyclization activity and higher hydrolysis and disproportionation activity | Paenibacillus macerans |
Q265K/Y195S | the mutant shows no cyclization (alpha-cyclodextrin-forming) and 226% hydrolysis (starch-degrading) activities compared to the wild type enzyme | Paenibacillus macerans |
Y195S | the mutant shows no cyclization (alpha-cyclodextrin-forming) and 200% hydrolysis (starch-degrading) activities compared to the wild type enzyme | Paenibacillus macerans |
Y195S | the mutant with enhanced maltodextrin specificity produces higher 2-O-D-glucopyranosyl-L-ascorbic acid yields than the wild type enzyme. Compared with the wild type enzyme, the mutant has lower cyclization activity and higher hydrolysis and disproportionation activity | Paenibacillus macerans |
Y195S/Q265K | compared with the wild type enzyme, the mutant has no cyclization activity and 498% hydrolysis and disproportionation activity | Paenibacillus macerans |
Y195S/Y260R | compared with the wild type enzyme, the mutant has lower cyclization activity and higher hydrolysis and disproportionation activity | Paenibacillus macerans |
Y260R | the mutant shows no cyclization (alpha-cyclodextrin-forming) and 226% hydrolysis (starch-degrading) activities compared to the wild type enzyme | Paenibacillus macerans |
Y260R | the mutant with enhanced maltodextrin specificity produces higher 2-O-D-glucopyranosyl-L-ascorbic acid yields than the wild type enzyme. Compared with the wild type enzyme, the mutant has lower cyclization activity and higher hydrolysis and disproportionation activity | Paenibacillus macerans |
Y260R/Q265K | compared with the wild type enzyme, the mutant has 8% cyclization activity and 213% hydrolysis activity | Paenibacillus macerans |
Y260R/Q265K/Y195S | the mutant shows 12% cyclization (alpha-cyclodextrin-forming) and 557% hydrolysis (starch-degrading) activities compared to the wild type enzyme | Paenibacillus macerans |
Y260R/Y195S | the mutant shows no cyclization (alpha-cyclodextrin-forming) and 492% hydrolysis (starch-degrading) activities compared to the wild type enzyme | Paenibacillus macerans |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | not inhibited by L-ascorbic acid | Paenibacillus macerans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.47 | - |
maltodextrin | mutant enzyme Y260R/Q265K/Y195S, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.48 | - |
maltodextrin | mutant enzyme Y260R/Q265K, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.5 | - |
maltodextrin | mutant enzyme Q265K/Y195S, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.51 | - |
maltodextrin | mutant enzyme Y260R, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.52 | - |
maltodextrin | mutant enzyme Q265K, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.53 | - |
maltodextrin | mutant enzyme Y260R/Y195S, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.62 | - |
maltodextrin | mutant enzyme Y195S, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.64 | - |
maltodextrin | wild type enzyme, at pH 5.5 and 37°C | Paenibacillus macerans | |
38.32 | - |
L-ascorbic acid | wild type enzyme, at pH 5.5 and 37°C | Paenibacillus macerans | |
43.44 | - |
L-ascorbic acid | mutant enzyme Y260R, at pH 5.5 and 37°C | Paenibacillus macerans | |
44.32 | - |
L-ascorbic acid | mutant enzyme Q265K, at pH 5.5 and 37°C | Paenibacillus macerans | |
44.52 | - |
L-ascorbic acid | mutant enzyme Y260R/Q265K, at pH 5.5 and 37°C | Paenibacillus macerans | |
47.21 | - |
L-ascorbic acid | mutant enzyme Q265K/Y195S, at pH 5.5 and 37°C | Paenibacillus macerans | |
48.47 | - |
L-ascorbic acid | mutant enzyme Y195S, at pH 5.5 and 37°C | Paenibacillus macerans | |
48.84 | - |
L-ascorbic acid | mutant enzyme Y260R/Y195S, at pH 5.5 and 37°C | Paenibacillus macerans | |
51.07 | - |
L-ascorbic acid | mutant enzyme Y260R/Q265K/Y195S, at pH 5.5 and 37°C | Paenibacillus macerans |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
75000 | - |
x * 75000, SDS-PAGE | Paenibacillus macerans |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Paenibacillus macerans | - |
- |
- |
Paenibacillus macerans JFB05-01 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA agarose column chromatography | Paenibacillus macerans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
maltodextrin + L-ascorbic acid | - |
Paenibacillus macerans | 2-O-D-glucopyranosyl-L-ascorbic acid + ? | - |
? | |
maltodextrin + L-ascorbic acid | - |
Paenibacillus macerans JFB05-01 | 2-O-D-glucopyranosyl-L-ascorbic acid + ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 75000, SDS-PAGE | Paenibacillus macerans |
Synonyms | Comment | Organism |
---|---|---|
CGTase | - |
Paenibacillus macerans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
36 | - |
- |
Paenibacillus macerans |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
the half-life at 40°C is 7.6 h | Paenibacillus macerans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000086 | - |
L-ascorbic acid | mutant enzyme Y195S, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.000097 | - |
L-ascorbic acid | wild type enzyme, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.0001 | - |
L-ascorbic acid | mutant enzyme Q265K, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.000105 | - |
L-ascorbic acid | mutant enzyme Y260R, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.000106 | - |
L-ascorbic acid | mutant enzyme Q265K/Y195S, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.00011 | - |
L-ascorbic acid | mutant enzyme Y260R/Q265K, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.000114 | - |
L-ascorbic acid | mutant enzyme Y260R/Q265K/Y195S, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.000133 | - |
L-ascorbic acid | mutant enzyme Y260R/Y195S, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.0059 | - |
maltodextrin | wild type enzyme, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.0067 | - |
maltodextrin | mutant enzyme Y195S, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.0084 | - |
maltodextrin | mutant enzyme Q265K, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.009 | - |
maltodextrin | mutant enzyme Y260R, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.0096 | - |
maltodextrin | mutant enzyme Q265K/Y195S, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.0098 | - |
maltodextrin | mutant enzyme Y260R/Y195S, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.0104 | - |
maltodextrin | mutant enzyme Y260R/Q265K, at pH 5.5 and 37°C | Paenibacillus macerans | |
0.0124 | - |
maltodextrin | mutant enzyme Y260R/Q265K/Y195S, at pH 5.5 and 37°C | Paenibacillus macerans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
- |
Paenibacillus macerans |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
5.5 | - |
the half-life at pH 5.5 is 29.8 h | Paenibacillus macerans |