Cloned (Comment) | Organism |
---|---|
expressed in Bacillus subitilis WB600 | Bacillus licheniformis |
Protein Variants | Comment | Organism |
---|---|---|
L134R | amyl, from recombinant Bacillus subtilis, when the pH is reduced from 6.5 to 4.5, compared with wild-type, Km increases but turnover number decreases, the mutants show an inverse trend, which results in catalytic efficiency (kcat/Km) increased. When the pH is 6.5, the kcat/Km is about 1.5times that of the mutants L134R, S320A, and L134R/S320A. In contrast, the kcat/Km of L134R and S320A are about 8.6- and 7.6times higher than that of the wild-type at pH 4.5. No significant difference on the kcat/Km of the mutants L134R, S320A, and L134R/S320A is shown when the pH is 5.5 and 6.5, respectively | Bacillus licheniformis |
L134R/S320A | amyd, from recombinant Bacillus subtilis, when the pH is reduced from 6.5 to 4.5, compared with wild-type, Km increases but turnover number decreases, the mutants show an inverse trend, which results in catalytic efficiency (kcat/Km) increased. The highest kcat/Km with pH 4.5 approximately 14times that of the wild-type is observed in the double mutant. No significant difference on the kcat/Km of the mutants L134R, S320A, and L134R/S320A is shown when the pH is 5.5 and 6.5, respectively | Bacillus licheniformis |
S320A | amy2, from recombinant Bacillus subtilis, when the pH is reduced from 6.5 to 4.5, compared with wild-type, Km increases but turnover number decreases, the mutants show an inverse trend, which results in catalytic efficiency (kcat/Km) increased. When the pH is 6.5, the kcat/Km is about 1.5times that of the mutants L134R, S320A, and L134R/S320A. In contrast, the kcat/Km of L134R and S320A are about 8.6- and 7.6times higher than that of the wild-type at pH 4.5. No significant difference on the kcat/Km of the mutants L134R, S320A, and L134R/S320A is shown when the pH is 5.5 and 6.5, respectively | Bacillus licheniformis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
starch | 0.60 mg/ml, double mutant, kcat/Km: 1560 ml/mg/s, L134R/S320A, 25°C, pH 6.0 | Bacillus licheniformis | |
additional information | - |
starch | 0.61 mg/ml, mutant S329A, kcat/Km: 1870 ml/mg/s, 25°C, pH 6.0 | Bacillus licheniformis | |
additional information | - |
starch | 0.64 mg/ml, mutant L134R, kcat/Km: 1670 ml/mg/s, 25°C, pH 6.0 | Bacillus licheniformis | |
additional information | - |
starch | 0.86 mg/ml, wild-type, kcat/Km: 1920 ml/mg/s, 25°C, pH 6.0 | Bacillus licheniformis | |
0.01 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | double mutant, kcat/Km: L134R/S320A, 680000 1/s/M, 25°C, pH 6.0 | Bacillus licheniformis | |
0.0113 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant S329A, kcat/Km: 740000 1/s/M, 25°C, pH 6.0 | Bacillus licheniformis | |
0.0117 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant L134R, kcat/Km: 710000 1/s/M, 25°C, pH 6.0 | Bacillus licheniformis | |
0.012 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | wild-type, kcat/Km: 770000 1/s/M, 25°C, pH 6.0 | Bacillus licheniformis | |
0.137 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | double mutant, L134R/S320A, kcat/Km: L134R/S320A, 1910000 1/s/M, 37°C, pH 4.5 | Bacillus licheniformis | |
0.147 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | wild-type, kcat/Km: 1420000 1/s/M, 37°C, pH 6.5 | Bacillus licheniformis | |
0.171 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant L134R, kcat/Km: 1350000 1/s/M, 37°C, pH 4.5 | Bacillus licheniformis | |
0.185 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | double mutant, L134R/S320A, kcat/Km: L134R/S320A, 680000 1/s/M, 37°C, pH 5.5 | Bacillus licheniformis | |
0.186 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant S329A, kcat/Km: 1210000 1/s/M, 37°C, pH 4.5 | Bacillus licheniformis | |
0.19 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | double mutant, L134R/S320A, kcat/Km: L134R/S320A, 1100000 1/s/M, 37°C, pH 6.5 | Bacillus licheniformis | |
0.193 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant L134R, kcat/Km: 1070000 1/s/M, 37°C, pH 5.5 | Bacillus licheniformis | |
0.2 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant S329A, kcat/Km: 1230000 1/s/M, 37°C, pH 5.5 | Bacillus licheniformis | |
0.201 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant L134R, kcat/Km: 920000 1/s/M, 37°C, pH 6.5 | Bacillus licheniformis | |
0.201 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | wild-type, kcat/Km: 870000 1/s/M, 37°C, pH 5.5 | Bacillus licheniformis | |
0.205 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant S329A, kcat/Km: 880000 1/s/M, 37°C, pH 6.5 | Bacillus licheniformis | |
0.281 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | wild-type, kcat/Km: 140000 1/s/M, 37°C, pH 4.5 | Bacillus licheniformis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus licheniformis | P06278 | wild-type; CICC 10181 | - |
Bacillus licheniformis CICC 10181 | P06278 | wild-type; CICC 10181 | - |
Purification (Comment) | Organism |
---|---|
wild-type, and recombinant mutants L134R, S320A, and L134R/S320A | Bacillus licheniformis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside + H2O | - |
Bacillus licheniformis | p-nitrophenol + 4,6-ethyliden-[G7]-alpha-D-maltoheptaoside | - |
? | |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside + H2O | - |
Bacillus licheniformis CICC 10181 | p-nitrophenol + 4,6-ethyliden-[G7]-alpha-D-maltoheptaoside | - |
? | |
starch + H2O | - |
Bacillus licheniformis | ? | - |
? | |
starch + H2O | - |
Bacillus licheniformis CICC 10181 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
alpha amylase | - |
Bacillus licheniformis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at, determination of catalytic properties on the substrates | Bacillus licheniformis |
37 | - |
assay at, determination of catalytic properties and pH study | Bacillus licheniformis |
70 | - |
assay at | Bacillus licheniformis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
100 | - |
around 60% of the enzyme activity is still detectable in wild-type and mutant enzymes after incubating in a boiling bath for 60 min | Bacillus licheniformis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
starch | double mutant, L134R/S320A, kcat/Km: 1560 ml/mg/s, L134R/S320A, 25°C, pH 6.0 | Bacillus licheniformis | |
additional information | - |
starch | mutant L134R, kcat/Km: 1670 ml/mg/s, 25°C, pH 6.0 | Bacillus licheniformis | |
additional information | - |
starch | mutant S329A, kcat/Km: 1870 ml/mg/s, 25°C, pH 6.0 | Bacillus licheniformis | |
additional information | - |
starch | wild-type, kcat/Km: 1920 ml/mg/s, 25°C, pH 6.0 | Bacillus licheniformis | |
7.86 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | double mutant, L134R/S320A, kcat/Km: L134R/S320A, 680000 1/s/M, 25°C, pH 6.0 | Bacillus licheniformis | |
8.14 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant L134R, kcat/Km: 710000 1/s/M, 25°C, pH 6.0 | Bacillus licheniformis | |
8.32 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant S329A, kcat/Km: 740000 1/s/M, 25°C, pH 6.0 | Bacillus licheniformis | |
8.73 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | wild-type, kcat/Km: 770000 1/s/M, 25°C, pH 6.0 | Bacillus licheniformis | |
34 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | wild-type, kcat/Km: 140000 1/s/M, 37°C, pH 4.5 | Bacillus licheniformis | |
175 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant S329A, kcat/Km: 880000 1/s/M, 37°C, pH 6.5 | Bacillus licheniformis | |
175 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | wild-type, kcat/Km: 870000 1/s/M, 37°C, pH 5.5 | Bacillus licheniformis | |
186 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant L134R, kcat/Km: 920000 1/s/M, 37°C, pH 6.5 | Bacillus licheniformis | |
187 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant S329A, kcat/Km: 1230000 1/s/M, 37°C, pH 5.5 | Bacillus licheniformis | |
201 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant L134R, kcat/Km: 1070000 1/s/M, 37°C, pH 5.5 | Bacillus licheniformis | |
205 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | wild-type, kcat/Km: 1420000 1/s/M, 37°C, pH 6.5 | Bacillus licheniformis | |
209 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | double mutant, L134R/S320A, kcat/Km: L134R/S320A, 1100000 1/s/M, 37°C, pH 6.5 | Bacillus licheniformis | |
219 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant S329A, kcat/Km: 1210000 1/s/M, 37°C, pH 4.5 | Bacillus licheniformis | |
233 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | double mutant, L134R/S320A, kcat/Km: L134R/S320A, 680000 1/s/M, 37°C, pH 5.5 | Bacillus licheniformis | |
234 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | mutant L134R, kcat/Km: 1350000 1/s/M, 37°C, pH 4.5 | Bacillus licheniformis | |
266 | - |
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside | double mutant, L134R/S320A, kcat/Km: L134R/S320A, 1910000 1/s/M, 37°C, pH 4.5 | Bacillus licheniformis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
assay at | Bacillus licheniformis |
7 | - |
optimum | Bacillus licheniformis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5.5 | 9.5 | 4000 U/ml at pH 5.5, above 4000 U/ml at pH 10.0, pH-optimum: 10000 U/ml | Bacillus licheniformis |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
4.5 | - |
the mutant L134R/S320A shows a stronger activity in acidic conditions, In contrast, the wild-type is sensitive to acidic pH. The double mutant is more effective in increasing enzyme stability against acidic pH than L134R and S320A | Bacillus licheniformis |