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Information on EC 3.2.1.1 - alpha-amylase and Organism(s) Hordeum vulgare and UniProt Accession P04063

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IUBMB Comments
Acts on starch, glycogen and related polysaccharides and oligosaccharides in a random manner; reducing groups are liberated in the alpha-configuration. The term "alpha" relates to the initial anomeric configuration of the free sugar group released and not to the configuration of the linkage hydrolysed.
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This record set is specific for:
Hordeum vulgare
UNIPROT: P04063
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Word Map
The taxonomic range for the selected organisms is: Hordeum vulgare
The enzyme appears in selected viruses and cellular organisms
Synonyms
alpha-amylase, diastase, alpha amylase, pancreatic alpha-amylase, crustacean cardioactive peptide, maltogenic amylase, taka-amylase a, human salivary alpha-amylase, bacillus licheniformis alpha-amylase, alpha-amylase 2, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
alpha-amylase 1
-
1,4-alpha-D-glucan glucanohydrolase
-
-
-
-
alpha amylase 1
-
-
Alpha-amylase carcinoid
-
-
-
-
alpha-amylase type A isozyme
-
alpha-amylases 1
-
Amy c6
-
-
-
-
Amylase THC 250
-
-
-
-
amylase, alpha-
-
-
-
-
Amylopsin
-
-
-
-
Bactosol TK
-
-
-
-
barley alpha-amylase 1
-
Buclamase
-
-
-
-
Clarase
-
-
-
-
Clone 103
-
-
-
-
Clone 168
-
-
-
-
Clone PHV19
-
-
-
-
Clones GRAMY56 and 963
-
-
-
-
diastase
-
-
-
-
endoamylase
-
-
-
-
Fortizyme
-
-
-
-
G 995
-
-
-
-
glycogenase
-
-
-
-
High pI alpha-amylase
-
-
-
-
Isozyme 1B
-
-
-
-
Kleistase L 1
-
-
-
-
Low pI alpha-amylase
Maxamyl
-
-
-
-
Maxilase
-
-
-
-
Meiotic expression upregulated protein 30
-
-
-
-
Pancreatic alpha-amylase
-
-
-
-
Pivozin
-
-
-
-
Ptyalin
-
-
-
-
Spitase CP 1
-
-
-
-
TAA
-
-
-
-
Taka-amylase A
-
-
-
-
Takatherm
-
-
-
-
Thermamyl
-
-
-
-
Thermolase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Endohydrolysis of (1->4)-alpha-D-glucosidic linkages in polysaccharides containing three or more (1->4)-alpha-linked D-glucose units
show the reaction diagram
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
SYSTEMATIC NAME
IUBMB Comments
4-alpha-D-glucan glucanohydrolase
Acts on starch, glycogen and related polysaccharides and oligosaccharides in a random manner; reducing groups are liberated in the alpha-configuration. The term "alpha" relates to the initial anomeric configuration of the free sugar group released and not to the configuration of the linkage hydrolysed.
CAS REGISTRY NUMBER
COMMENTARY hide
9000-90-2
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-chloro-4-nitrophenyl beta-D-maltoheptaoside + H2O
2-chloro-4-nitrophenol + beta-D-maltoheptaose
show the reaction diagram
-
-
-
?
amylose + H2O
oligosaccharides
show the reaction diagram
amylose DP17, preferred substrate of mutant enzyme T212Y
-
-
?
amylose DP17 + H2O
?
show the reaction diagram
-
-
-
?
beta-cyclodextrin + H2O
?
show the reaction diagram
insight into the action of alpha-amylase at the molecular level
-
-
?
starch + H2O
oligosaccharides
show the reaction diagram
insoluble blue starch, preferred substrate of mutant enzyme Y105A
-
-
?
2-chloro-4-nitrophenyl alpha-D-maltoheptaoside + H2O
2-chloro-4-nitrophenol + alpha-D-maltoheptaose
show the reaction diagram
-
-
-
-
?
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 + ?
show the reaction diagram
pH 6.8, 30°C
-
-
?
2-chloro-4-nitrophenyl beta-D-maltoheptaoside + H2O
?
show the reaction diagram
4,6-ethylidene-[G7]-p-nitrophenyl-[G1]-alpha-D-maltoheptaoside + H2O
p-nitrophenol + 4,6-ethyliden-[G7]-alpha-D-maltoheptaoside
show the reaction diagram
-
-
-
-
?
amylose + H2O
?
show the reaction diagram
amylose DP440, pH 6.8, 30°C
-
-
?
amylose + H2O
malto-oligosaccharides
show the reaction diagram
amylose + H2O
maltooligosaccharides
show the reaction diagram
-
-
-
-
?
amylose DP17 + H2O
?
show the reaction diagram
-
-
-
-
?
amylose DP440 + H2O
?
show the reaction diagram
beta-cyclodextrin + H2O
?
show the reaction diagram
insight into the action of alpha-amylase at the molecular level
-
-
?
insoluble Blue Starch + H2O
?
show the reaction diagram
pH 5.5, 37°C
-
-
?
maltodextrin + H2O
maltooligosaccharides
show the reaction diagram
-
-
-
-
?
maltoheptaose + H2O
maltohexaose + maltopentaose + maltotetraose + maltotriose + maltose
show the reaction diagram
-
-
-
-
?
maltoheptaose + H2O
maltotriose + maltose + D-glucose
show the reaction diagram
-
-
identified by thin-layer-chromatography
-
?
maltohexaose + H2O
maltotetraose + maltotriose + maltose + D-glucose
show the reaction diagram
-
-
identified by thin-layer-chromatography
-
?
maltopentaose + H2O
maltotetraose + maltotriose + maltose + D-glucose
show the reaction diagram
-
-
identified by thin-layer-chromatography
-
?
maltotetraose + H2O
maltotriose + maltose
show the reaction diagram
-
-
identified by thin-layer-chromatography
-
?
maltotriose + H2O
maltose + D-glucose
show the reaction diagram
-
-
identified by thin-layer-chromatography
-
?
p-nitrophenyl maltoheptaoside + H2O
?
show the reaction diagram
-
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
?
show the reaction diagram
-
-
-
-
?
starch + H2O
malto-oligosaccharides
show the reaction diagram
starch + H2O
maltooligosaccharides
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
amylose + H2O
malto-oligosaccharides
show the reaction diagram
-
-
-
-
?
amylose + H2O
maltooligosaccharides
show the reaction diagram
-
-
-
-
?
starch + H2O
malto-oligosaccharides
show the reaction diagram
-
-
-
-
?
starch + H2O
maltooligosaccharides
show the reaction diagram
-
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
alpha-cyclodextrin
-
weak inhibition
O-4,6-dideoxy-4-{[4,5,6-trihydroxy-3-hydroxymethyl-2-cyclohexen-1-yl]amino}-alpha-D-glucopyranosyl-(1-4)-O-alpha-D-glucopyranosyl-(1-4)-D-glucose
-
trivial name acarbose, uncompetitive inhibition vs. amylose and maltodextrin, mixed noncompetitive inhibition vs. maltoheptaose
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
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
0.00016 - 0.45
amylose
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
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
10 - 203
2-chloro-4-nitrophenyl beta-D-maltoheptaoside
78 - 264
amylose DP17
-
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
56 - 206
amylose
225
amylose DP17
-
pH 5.5, 37°C, recombinant AMY1-SBD fusion enzyme
-
95 - 209
amylose DP440
-
125 - 129
maltodextrin
2 - 5.6
maltoheptaose
additional information
additional information
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1900
-
recombinant Amy1, hydrolysis of soluble starch at 37°C and pH 6.0
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.74
-
recombinant AMY1-SBD fusion enzyme, amino acid sequence calculation
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
SwissProt
Manually annotated by BRENDA team
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
aleurone layer, two alpha-amylase families, the type A mRNAs are present in relative large amounts in unstimulated aleurone cells and increase about 20fold after stimulation with giberellic acid, type B mRNA is present at very low levels in unstimulated cells, but increases at least 100fold after cells are exposed to the hormone
Manually annotated by BRENDA team
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
AMY2_HORVU
427
1
47356
Swiss-Prot
Secretory Pathway (Reliability: 1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45000
68900
-
x * 68900, recombinant AMY1-SBD fusion enzyme, amino acid sequence calculation, x * 75000, recombinant glycosylated AMY1-SBD fusion enzyme, SDS-PAGE
75000
-
x * 68900, recombinant AMY1-SBD fusion enzyme, amino acid sequence calculation, x * 75000, recombinant glycosylated AMY1-SBD fusion enzyme, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
proteolytic modification
-
types of processing of barley rAMY1 produced in yeast: 1. processing of the signal peptide, 2. removal of the C-terminal Arg-Ser, 3. glutathionylation of Cys95, 4. O-glycosylation, 5. additional degradation of the C-terminus
additional information
-
types of processing of barley rAMY1 produced in yeast: 1. processing of the signal peptide, 2. removal of the C-terminal Arg-Ser, 3. glutathionylation of Cys95, 4. O-glycosylation, 5. additional degradation of the C-terminus
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
mutants Y105A/Y380A and H395A are crystallized
the structures of AMY1 Y380A and S378P are compared with the wild-type enzyme both in free form and in complex with acarbose
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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
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
Y380M
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
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5
-
AMY1 is more stable at low pH than AMY2. The activity of AMY2 rapidly decreases to 51% after 3 min incubation at pH 3.5
693604
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Ca2+ stabilizes under combined pressure–temperature treatments in the range of 0.1–800 MPa and 30–75°C
-
pressure up to 200 MPa significantly stabilizs the enzyme against temperature-induced inactivation
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, concentrated purified recombinant AMY1-SBD fusion enzyme, 50 mM MES, pH 6.5, 5 mM CaCl2, stable
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant wild-type and mutant enzymes from Pichia pastoris strain GS115 by beta-cyclodextrin affinity chromatography
purified from green and kilned malt
-
recombinant Amy1, affinity chromatography on cyclohepta-amylase substituted epoxy-Sepharose 6B
-
recombinant AMY1-SBD fusion enzyme from Aspergillus niger strain AB4.1 by beta-cyclodextrin affinity chromatography
-
recombinant enzyme
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
AMY1, DNA and amino acid sequence determination and analysis, cloning in Escherichia coli strain DH5alpha, expression of wild-type and mutant enzymes in Pichia pastoris strain GS115
AMY1, subcloning in Escherichia coli strain DH5alpha, expression of the alpha-amylase C-terminally fused to the Aspergillus niger glucoamlyase starch binding domain in Aspergillus niger strain AB4.1, the mutant enzyme is secreted to the culture medium due to the signal peptide of the barley alpha-amylase
-
cloned and overexpressed in Pichia pastoris, recombinant isoenzymes with a six-histidine fusion tag
-
cloned in Escherichia coli DH5alpha and XL10-Gold to pPICZA-AMY1. pPICZA-AMY1 and the mutants Y105A/Y380A, Y105A/Y380M, H395A, Y380A/H395A, Y380A, Y380M, and Y105A are expressed in Pichia pastoris
expression in Pichia pastoris
expression in Saccharomyces cerevisiae
-
secreted and correctly processed by Aspergillus niger
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
nutrition
-
the enzyme is crucial for the production of malt, an imortant starting material in the manufacture of beer and whisky
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Rogers, J.C.
Two barley alpha-amylase gene families are regulated differently in aleurone cells
J. Biol. Chem.
260
3731-3738
1985
Hordeum vulgare
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team
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)
-
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team