Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
2-chloro-4-nitrophenyl beta-D-maltoheptaoside + H2O
2-chloro-4-nitrophenol + beta-D-maltoheptaose
-
-
-
?
amylose + H2O
oligosaccharides
amylose DP17, preferred substrate of mutant enzyme T212Y
-
-
?
amylose DP17 + H2O
?
-
-
-
?
beta-cyclodextrin + H2O
?
insight into the action of alpha-amylase at the molecular level
-
-
?
starch + H2O
oligosaccharides
insoluble blue starch, preferred substrate of mutant enzyme Y105A
-
-
?
2-chloro-4-nitrophenyl alpha-D-maltoheptaoside + H2O
2-chloro-4-nitrophenol + alpha-D-maltoheptaose
-
-
-
-
?
2-chloro-4-nitrophenyl beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranoside + H2O
2-chloro-4-nitrophenol + ?
pH 6.8, 30°C
-
-
?
2-chloro-4-nitrophenyl beta-D-maltoheptaoside + H2O
?
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside + H2O
p-nitrophenol + 4,6-ethyliden-[G7]-alpha-D-maltoheptaoside
-
-
-
-
?
amylose + H2O
?
amylose DP440, pH 6.8, 30°C
-
-
?
amylose + H2O
malto-oligosaccharides
amylose + H2O
maltooligosaccharides
-
-
-
-
?
amylose DP17 + H2O
?
-
-
-
-
?
beta-cyclodextrin + H2O
?
insight into the action of alpha-amylase at the molecular level
-
-
?
insoluble Blue Starch + H2O
?
pH 5.5, 37°C
-
-
?
maltodextrin + H2O
maltooligosaccharides
-
-
-
-
?
maltoheptaose + H2O
maltohexaose + maltopentaose + maltotetraose + maltotriose + maltose
-
-
-
-
?
maltoheptaose + H2O
maltotriose + maltose + D-glucose
-
-
identified by thin-layer-chromatography
-
?
maltohexaose + H2O
maltotetraose + maltotriose + maltose + D-glucose
-
-
identified by thin-layer-chromatography
-
?
maltopentaose + H2O
maltotetraose + maltotriose + maltose + D-glucose
-
-
identified by thin-layer-chromatography
-
?
maltotetraose + H2O
maltotriose + maltose
-
-
identified by thin-layer-chromatography
-
?
maltotriose + H2O
maltose + D-glucose
-
-
identified by thin-layer-chromatography
-
?
p-nitrophenyl maltoheptaoside + H2O
?
-
a maximum of substrate cleavage was identified at 152 MPa and 64°C, yielding approximately 25% higher substrate conversion after 30 min, as compared to the maximum at ambient pressure and 59°C
-
-
?
starch + H2O
malto-oligosaccharides
starch + H2O
maltooligosaccharides
-
-
-
-
?
additional information
?
-
2-chloro-4-nitrophenyl beta-D-maltoheptaoside + H2O
?
-
-
-
-
?
2-chloro-4-nitrophenyl beta-D-maltoheptaoside + H2O
?
-
-
-
?
amylose + H2O
malto-oligosaccharides
-
-
-
-
?
amylose + H2O
malto-oligosaccharides
-
amylose DP440 and amylose DP17
-
-
?
amylose DP440 + H2O
?
-
-
-
-
?
amylose DP440 + H2O
?
-
-
-
?
starch + H2O
malto-oligosaccharides
-
-
-
-
?
starch + H2O
malto-oligosaccharides
-
starch granules, high affinity for the substrate is mediated by the enzyme's separate starch binding domain, SBD
-
-
?
additional information
?
-
substrate specificities of wild-type and mutant enzymes, oligosaccharide action patterns of mutant enzymes, overview, Tyr105 and Thr212 at outermost substrate binding subsites -6 and +4 control substrate specificity, oligosaccharide cleavage patterns, and multiple binding modes of alpha-amylase 1, modelling of oligosaccharide substrate docking at the substrate binding area
-
-
?
additional information
?
-
-
substrate specificities of wild-type and mutant enzymes, oligosaccharide action patterns of mutant enzymes, overview, Tyr105 and Thr212 at outermost substrate binding subsites -6 and +4 control substrate specificity, oligosaccharide cleavage patterns, and multiple binding modes of alpha-amylase 1, modelling of oligosaccharide substrate docking at the substrate binding area
-
-
?
amylose + H2O
additional information
-
-
short chain, DP 17
-
-
?
starch + H2O
additional information
-
-
-
-
-
?
starch + H2O
additional information
-
-
soluble starch
-
-
?
starch + H2O
additional information
-
-
insoluble starch
-
-
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
0.6 - 10
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
0.12 - 2.36
amylose DP17
-
0.3 - 1.5
insoluble blue starch
-
2.3
2-chloro-4-nitrophenyl alpha-D-maltoheptaoside
-
pH 6.8, 30°C, recombinant AMY1-SBD fusion enzyme
0.669 - 10
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
0.45
4,6-ethylidene-[G7]-p-nitrophenyl[G1]-alpha-D-maltoheptaoside
-
37°C, pH 6.0
0.37
amylose DP17
-
pH 5.5, 37°C, recombinant AMY1-SBD fusion enzyme
-
0.23
amylose DP440
-
pH 5.5, 37°C, recombinant AMY1-SBD fusion enzyme
-
0.071 - 0.079
maltodextrin
0.21 - 0.22
maltoheptaose
additional information
amylose
0.6
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, mutant T212W
1.3
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, mutant Y105W
1.4
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, mutant T212P
1.7
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, wild-type enzyme
2
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, mutant T212Y
3.1
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, mutant Y105A/T212W
4.9
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, mutant Y105F
6
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, mutant Y105A/T212Y
10
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
above, pH 5.5, 37°C, mutant Y105A
0.12
amylose DP17
pH 5.5, 37°C, mutant T212Y
-
0.29
amylose DP17
pH 5.5, 37°C, mutant T212W
-
0.48
amylose DP17
pH 5.5, 37°C, mutants Y105W and T212P
-
0.57
amylose DP17
pH 5.5, 37°C, wild-type enzyme
-
0.58
amylose DP17
pH 5.5, 37°C, mutant Y105F
-
2
amylose DP17
pH 5.5, 37°C, mutant Y105A/T212W
-
2.27
amylose DP17
pH 5.5, 37°C, mutant Y105A/T212Y
-
2.36
amylose DP17
below, pH 5.5, 37°C, mutant Y105A
-
0.3
insoluble blue starch
pH 5.5, 37°C, mutant T212W
-
0.3
insoluble blue starch
pH 5.5, 37°C, mutant Y105A/T212W
-
0.4
insoluble blue starch
below, pH 5.5, 37°C, mutant Y105A
-
0.4
insoluble blue starch
pH 5.5, 37°C, mutant Y105F
-
0.4
insoluble blue starch
pH 5.5, 37°C, wild-type enzyme
-
0.5
insoluble blue starch
pH 5.5, 37°C, mutant Y105W
-
0.6
insoluble blue starch
pH 5.5, 37°C, mutant T212Y
-
1
insoluble blue starch
pH 5.5, 37°C, mutant Y105A/T212Y
-
1.5
insoluble blue starch
pH 5.5, 37°C, mutant T212P
-
0.669
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
mutant enzyme Y380A
0.67
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
Y380A mutant, pH 5.5, 30°C, kcat/Km: 28 1/s/mg/ml
0.7
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
wild-type enzyme, pH 5.5, 30°C, kcat/Km: 38 1/s/mg/ml
0.724
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
mutant enzyme Y380F
0.735
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
mutant enzyme S378T
0.758
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
alpha-amylase 1
0.861
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
mutant enzyme S378P
0.87
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
Y380M mutant, pH 5.5, 30°C, kcat/Km: 39 1/s/mg/ml
0.871
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
mutant enzyme Y380M
0.91
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
H395A mutant, kcat below 10 1/s, pH 5.5, 30°C
1.16
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
-
malt enzyme
1.4
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
Y380A/H395A mutant, pH 5.5, 30°C, kcat/Km: 25 1/s/mg/ml
2.49
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
-
recombinant enzyme
10
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
Y105A mutant, Km above 10 mM, kcat below 10 1/s, pH 5.5, 30°C
10
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
Y105A/Y380A mutant, Km above 10 mM, pH 5.5, 30°C, kcat/Km: 1300 1/s/mg/ml
10
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
Y105A/Y380M mutant, Km above 10 mM, kcat below 10 1/s, pH 5.5, 30°C
0.00016
amylose
-
30°C, pH 5.5
0.00021
amylose
-
30°C, pH 5.5
0.38
amylose
-
DP 17, malt enzyme, calculated as low molecular weight products
0.45
amylose
-
DP 17, recombinant enzyme, calculated as low molecular weight products
0.071
maltodextrin
-
30°C, pH 5.5
0.079
maltodextrin
-
30°C, pH 5.5
0.21
maltoheptaose
-
30°C, pH 5.5
0.22
maltoheptaose
-
30°C, pH 5.5
additional information
amylose
0.13 mg/ml, wild-type enzyme, pH 5.5, 30°C, kcat/Km: 1300 1/s/mg/ml
additional information
amylose
-
0.13 mg/ml, wild-type enzyme, pH 5.5, 30°C, kcat/Km: 1300 1/s/mg/ml
additional information
amylose
0.19 mg/ml, H395A mutant, pH 5.5, 30°C, kcat/Km: 394 1/s/mg/ml
additional information
amylose
-
0.19 mg/ml, H395A mutant, pH 5.5, 30°C, kcat/Km: 394 1/s/mg/ml
additional information
amylose
0.30 mg/ml, Y380A/H395A mutant, pH 5.5, 30°C, kcat/Km: 543 1/s/mg/ml
additional information
amylose
-
0.30 mg/ml, Y380A/H395A mutant, pH 5.5, 30°C, kcat/Km: 543 1/s/mg/ml
additional information
amylose
0.35 mg/ml, Y380M mutant, pH 5.5, 30°C, kcat/Km: 425 1/s/mg/ml
additional information
amylose
-
0.35 mg/ml, Y380M mutant, pH 5.5, 30°C, kcat/Km: 425 1/s/mg/ml
additional information
amylose
0.36 mg/ml, Y380A mutant, pH 5.5, 30°C, kcat/Km: 264 1/s/mg/ml
additional information
amylose
-
0.36 mg/ml, Y380A mutant, pH 5.5, 30°C, kcat/Km: 264 1/s/mg/ml
additional information
amylose
1.0 mg/ml, Y105A/Y380M mutant, pH 5.5, 30°C, kcat/Km: 56 1/s/mg/ml
additional information
amylose
-
1.0 mg/ml, Y105A/Y380M mutant, pH 5.5, 30°C, kcat/Km: 56 1/s/mg/ml
additional information
amylose
1.2 mg/ml, Y105A/Y380A mutant, pH 5.5, 30°C, kcat/Km: 51 1/s/mg/ml
additional information
amylose
-
1.2 mg/ml, Y105A/Y380A mutant, pH 5.5, 30°C, kcat/Km: 51 1/s/mg/ml
additional information
amylose
1.4 mg/ml, Y105A mutant, pH 5.5, 30°C, kcat/Km: 132 1/s/mg/ml
additional information
amylose
-
1.4 mg/ml, Y105A mutant, pH 5.5, 30°C, kcat/Km: 132 1/s/mg/ml
additional information
additional information
-
kinetics
-
additional information
additional information
-
Km-values of wild-type and mutant enzymes
-
additional information
additional information
KM-values for amylose: 0.363 mg/ml for mutant enzyme Y380A, 0.351 mg/ml for mutant enzyme Y380M, 0.391 mg/ml for mutant enzyme Y380F, 0.203 mg/ml for mutant enzyme S378P, 0.208 mg/ml for mutant enzyme S378T, 0.19 mg/ml for alpha-amylase 1
-
additional information
additional information
-
KM-values for amylose: 0.363 mg/ml for mutant enzyme Y380A, 0.351 mg/ml for mutant enzyme Y380M, 0.391 mg/ml for mutant enzyme Y380F, 0.203 mg/ml for mutant enzyme S378P, 0.208 mg/ml for mutant enzyme S378T, 0.19 mg/ml for alpha-amylase 1
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
10 - 203
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
132
2-chloro-4-nitrophenyl alpha-D-maltoheptaoside
-
pH 6.8, 30°C, recombinant AMY1-SBD fusion enzyme
3 - 35
2-chloro-4-nitrophenyl beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranoside
19 - 63.1
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
120
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside
-
37°C, pH 6.0
225
amylose DP17
-
pH 5.5, 37°C, recombinant AMY1-SBD fusion enzyme
-
additional information
additional information
-
10
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
below, pH 5.5, 37°C, mutant Y105A
24
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, mutant Y105A/T212W
31
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, mutant Y105A/T212Y
61
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, mutant T212W
119
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, wild-type enzyme
127
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, mutant T212Y
132
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, mutant Y105W
140
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, mutant T212P
203
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
pH 5.5, 37°C, mutant Y105F
78
amylose DP17
pH 5.5, 37°C, mutant Y105A/T212Y
-
105
amylose DP17
pH 5.5, 37°C, mutant Y105A/T212W
-
127
amylose DP17
pH 5.5, 37°C, mutant T212Y
-
146
amylose DP17
below, pH 5.5, 37°C, mutant Y105A
-
154
amylose DP17
pH 5.5, 37°C, mutant T212W
-
158
amylose DP17
pH 5.5, 37°C, mutant Y105F
-
165
amylose DP17
pH 5.5, 37°C, wild-type enzyme
-
200
amylose DP17
pH 5.5, 37°C, mutant Y105W
-
264
amylose DP17
pH 5.5, 37°C, mutant T212P
-
3 - 6
2-chloro-4-nitrophenyl beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranoside
wild-type enzyme, pH 5.5, 30°C, kcat/Km: 48 1/s/mg/ml
10
2-chloro-4-nitrophenyl beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranoside
Y105A mutant, kcat below 10 1/s, pH 5.5, 30°C
10
2-chloro-4-nitrophenyl beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranoside
Y105A/Y380A mutant, kcat below 10 1/s, pH 5.5, 30°C
10
2-chloro-4-nitrophenyl beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranoside
Y105A/Y380M mutant, kcat below 10 1/s, pH 5.5, 30°C
19
2-chloro-4-nitrophenyl beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranoside
Y380A mutant, pH 5.5, 30°C, kcat/Km: 28 1/s/mg/ml
21
2-chloro-4-nitrophenyl beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranoside
H395A mutant, pH 5.5, 30°C, kcat/Km: 23 1/s/mg/ml
34
2-chloro-4-nitrophenyl beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranoside
Y380M mutant, pH 5.5, 30°C, kcat/Km: 39 1/s/mg/ml
35
2-chloro-4-nitrophenyl beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranoside
Y380A/H395A mutant, pH 5.5, 30°C, kcat/Km: 25 1/s/mg/ml
19
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
mutant enzyme Y380A
34
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
mutant enzyme Y380M
40.4
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
-
malt enzyme
48
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
mutant enzyme S378T
52
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
alpha-amylase 1
56
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
mutant enzyme Y380F
59
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
mutant enzyme S378P
63.1
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
-
recombinant enzyme
56
amylose
Y105A/Y380M mutant, pH 5.5, 30°C, kcat/Km: 56 1/s/mg/ml, kcat/Km decreases 10-25fold for all Y105 mutants
61
amylose
Y105A/Y380A mutant, pH 5.5, 30°C, kcat/Km: 51 1/s/mg/ml, kcat/Km decreases 10-25fold for all Y105 mutants
75
amylose
H395A mutant, pH 5.5, 30°C, kcat/Km: 394 1/s/mg/ml
95
amylose
Y380A mutant, pH 5.5, 30°C, kcat/Km: 264 1/s/mg/ml
144
amylose
-
DP 17, malt enzyme, calculated as low molecular weight products
149
amylose
Y380M mutant, pH 5.5, 30°C, kcat/Km: 425 1/s/mg/ml
163
amylose
Y380A/H395A mutant, pH 5.5, 30°C, kcat/Km: 543 1/s/mg/ml
169
amylose
wild-type enzyme, pH 5.5, 30°C, kcat/Km: 1300 1/s/mg/ml
178
amylose
Y105A mutant, pH 5.5, 30°C, kcat/Km: 132 1/s/mg/ml, kcat/Km decreases 10-25fold for all Y105 mutants
185
amylose
-
DP 17, recombinant enzyme, calculated as low molecular weight products
202
amylose
-
30°C, pH 5.5
206
amylose
-
30°C, pH 5.5
95
amylose DP440
mutant enzyme Y380A
-
144
amylose DP440
mutant enzyme S378T
-
149
amylose DP440
mutant enzyme Y380M
-
162
amylose DP440
mutant enzyme Y380F
-
163
amylose DP440
mutant enzyme S378P
-
185
amylose DP440
alpha-amylase 1
-
209
amylose DP440
-
pH 5.5, 37°C, recombinant AMY1-SBD fusion enzyme
-
125
maltodextrin
-
30°C, pH 5.5
129
maltodextrin
-
30°C, pH 5.5
2
maltoheptaose
-
30°C, pH 5.5
5.6
maltoheptaose
-
30°C, pH 5.5
additional information
additional information
-
-
-
additional information
additional information
-
KM-values of wild-type and mutant enzymes
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
T212P
site-directed mutagenesis, the mutant shows altered substrate specificity and kinetics compared to the wild-type enzyme
T212W
site-directed mutagenesis, the mutant shows altered substrate specificity and kinetics compared to the wild-type enzyme
T212Y
site-directed mutagenesis, the mutant shows altered substrate specificity and kinetics compared to the wild-type enzyme
Y105A
site-directed mutagenesis, the mutant shows altered substrate specificity and kinetics compared to the wild-type enzyme
Y105A/T212W
site-directed mutagenesis, the mutant shows altered substrate specificity and kinetics compared to the wild-type enzyme
Y105A/T212Y
site-directed mutagenesis, the mutant shows altered substrate specificity and kinetics compared to the wild-type enzyme
Y105F
site-directed mutagenesis, the mutant shows altered substrate specificity and kinetics compared to the wild-type enzyme
Y105W
site-directed mutagenesis, the mutant shows altered substrate specificity and kinetics compared to the wild-type enzyme
H395A
located in the C-domain, the sugar tong, may be involved in the allosteric activation of the enzyme
S378P
kcat/KM for amylose is 1.2fold lower than wild-type value. kcat/KM for 2-chloro-4-nitrophenyl beta-D-maltoheptaoside is nearly identical to wild-type value
S378T
kcat/KM for amylose is 1.4fold lower than wild-type value. kcat/KM for 2-chloro-4-nitrophenyl beta-D-maltoheptaoside is nearly identical to wild-type value
Y105A
43% of the activity compared to wild-type
Y105A/Y380A
loss of 18-36% activity relative to wild-type
Y105A/Y380M
loss of 18-36% activity relative to wild-type
Y380A/H395A
located in the C-domain, the sugar tong, may be involved in the allosteric activation of the enzyme
Y380F
kcat/KM for amylose is fold lowerthan wild-type value. kcat/KM for is 2.6fold lower than wild-type value. kcat/KM for 2-chloro-4-nitrophenyl beta-D-maltoheptaoside is 1.1fold higher than wild-type value
additional information
-
construction of a mutant alpha-amylase, containing its signal peptide, which is fused to the starch binding domain, SBD, of the glucoamylase GA-I of Aspergillus niger via a 37 amino acid GA-I linker segment, the activity of the fusion protein is 2fold enhanced with amylose, and with starch at low concentration, not at high concentration, compared to the wild-type enzyme
Y380A
mutant fails to bind to beta-cyclodextrin-Sepharose, a starch-mimic resin used for alpha-amylase affinity purification. The Kd for beta-cyclodextrin binding to Y380A is 1.4 mm compared to 0.20-0.25 mM for the wild-type, S378P and S378T enzymes. Crystal structures of both wild-type and S378P enzymes, but not the Y380A enzyme, shows binding of the pseudotetrasaccharide acarbose at the sugar tongs site. beta-Cyclodextrin both inhibits binding to and suppresses activity on starch granules for wild-type and S378P enzymes, but does not affect these properties of Y380A. Y380A enzyme hydrolyzes amylose with reduced multiple attack. kcat/KM for amylose is 3.7fold lower than wild-type value. kcat/KM for 2-chloro-4-nitrophenyl beta-D-maltoheptaoside is 2.4fold lower than wild-type value
Y380A
29-50% decrease in activity compared to wild-type enzyme
Y380M
mutant fails to bind to beta-cyclodextrin-Sepharose, a starch-mimic resin used for alpha-amylase affinity purification. The Kd for beta-cyclodextrin binding to Y380M is 1.4 mm compared to 0.20-0.25 mM for the wild-type, S378P and S378T enzymes. kcat/KM for amylose is 2.3fold lower than wild-type value. kcat/KM for 2-chloro-4-nitrophenyl beta-D-maltoheptaoside is 1.8fold lower than wild-type value
Y380M
29-50% decrease in activity compared to wild-type enzyme
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Rogers, J.C.
Two barley alpha-amylase gene families are regulated differently in aleurone cells
J. Biol. Chem.
260
3731-3738
1985
Hordeum vulgare
brenda
Svensson, B.
Protein engineering in the alpha-amylase family: catalytic mechanism, substrate specificity and stability
Plant Mol. Biol.
25
141-157
1994
Aspergillus oryzae, Geobacillus stearothermophilus, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus licheniformis, Drosophila melanogaster, Escherichia coli, Saccharomycopsis fibuligera, Homo sapiens, Hordeum vulgare, Mus musculus, Oryza sp., Vigna radiata, Streptomyces hygroscopicus, Sus scrofa, Triticum aestivum, Xanthomonas campestris
brenda
Juge, N.; Svensson, B.; Williamson, G.
Secretion, purification, and characterisation of barley alpha-amylase produced by heterologous gene expression in Aspergillus niger
Appl. Microbiol. Biotechnol.
49
385-392
1998
Hordeum vulgare
brenda
Sogaard, M.; Andersen, J.S.; Roepstorff, R.; Svensson, B.
Electrospray mass spectrometry characterization of post-translational modification of barley alpha-amylase 1 produced in yeast
Biotechnology
11
1162-1165
1993
Hordeum vulgare
brenda
Oudjeriouat, N.; Moreau, Y.; Santimone, M.; Svensson, B.; Marchis-Mouren, G.; Desseaux, V.
On the mechanism of alpha-amylase
Eur. J. Biochem.
270
3871-3879
2003
Hordeum vulgare
brenda
Wong, D.W.S.; Batt, S.B.; Robertson, G.H.
Characterization of active barley alpha-amylase 1 expressed and secreted by Saccharomyces cerevisiae
J. Protein Chem.
20
619-623
2002
Hordeum vulgare
brenda
Juge, N.; Nohr, J.; Le Gal-Coeffet, M.F.; Kramhoft, B.; Furniss, C.S.; Planchot, V.; Archer, D.B.; Williamson, G.; Svensson, B.
The activity of barley alpha-amylase on starch granules is enhanced by fusion of a starch binding domain from Aspergillus niger glucoamylase
Biochim. Biophys. Acta
1764
275-284
2006
Hordeum vulgare
brenda
Bak-Jensen, K.S.; Andre, G.; Gottschalk, T.E.; Paes, G.; Tran, V.; Svensson, B.
Tyrosine 105 and threonine 212 at outermost substrate binding subsites -6 and +4 control substrate specificity, oligosaccharide cleavage patterns, and multiple binding modes of barley alpha-amylase 1
J. Biol. Chem.
279
10093-10102
2004
Hordeum vulgare (P04063), Hordeum vulgare
brenda
Buckow, R.; Weiss, U.; Heinz, V.; Knorr, D.
Stability and catalytic activity of alpha-amylase from barley malt at different pressure-temperature conditions
Biotechnol. Bioeng.
97
1-11
2007
Hordeum vulgare
brenda
Bozonnet, S.; Jensen, M.T.; Nielsen, M.M.; Aghajari, N.; Jensen, M.H.; Kramhoft, B.; Willemoes, M.; Tranier, S.; Haser, R.; Svensson, B.
The pair of sugar tongs site on the non-catalytic domain C of barley alpha-amylase participates in substrate binding and activity
FEBS J.
274
5055-5067
2007
Hordeum vulgare (P00693), Hordeum vulgare
brenda
Hachem, M.A.; Bozonnet, S.; Willemoes, M.; Bonsager, B.C.; Nielsen, M.M.; Fukuda, K.; Kramhoft, B.; Maeda, K.; Sigurskjold, B.W.; Hagglund, P.; Finnie, C.; Mori, H.; Robert, X.; Jensen, M.H.; Tranier, S.; Aghajari, N.; Haser, R.; Svensoon, B.
Interactions between barley alpha-amylases, substrates, inhibitors and regulatory proteins
J. Appl. Glycosci.
53
163-169
2006
Hordeum vulgare (P00693), Hordeum vulgare (P04063)
-
brenda
Nielsen, M.M.; Seo, E.S.; Bozonnet, S.; Aghajari, N.; Robert, X.; Haser, R.; Svensson, B.
Multi-site substrate binding and interplay in barley alpha-amylase 1
FEBS Lett.
582
2567-2571
2008
Hordeum vulgare (P00693), Hordeum vulgare
brenda
Yuk, J.B.; Choi, S.H.; Lee, T.H.; Jang, M.U.; Park, J.M.; Yi, A.R.; Svensson, B.; Kim, T.J.
Effects of calcium ion concentration on starch hydrolysis of barley alpha-amylase isozymes
J. Microbiol. Biotechnol.
18
730-734
2008
Hordeum vulgare
brenda