Information on EC 3.2.1.68 - isoamylase

New: Word Map on EC 3.2.1.68
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Search Reference ID:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea

EC NUMBER
COMMENTARY hide
3.2.1.68
-
RECOMMENDED NAME
GeneOntology No.
isoamylase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
hydrolysis of (1->6)-alpha-D-glucosidic branch linkages in glycogen, amylopectin and their beta-limit dextrins
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
metabolism of disaccharids
-
-
starch biosynthesis
-
-
starch degradation II
-
-
trehalose biosynthesis V
-
-
SYSTEMATIC NAME
IUBMB Comments
glycogen 6-alpha-D-glucanohydrolase
Also readily hydrolyses amylopectin. Differs from EC 3.2.1.41 (pullulanase) and EC 3.2.1.142 (limit dextrinase) by its inability to hydrolyse pullulan, and by limited action on alpha-limit dextrins. Maltose is the smallest sugar it can release from an alpha-(1->6)-linkage.
CAS REGISTRY NUMBER
COMMENTARY hide
9067-73-6
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
gene ArDBE
UniProt
Manually annotated by BRENDA team
gene treX, component of a treXYZ operon
-
-
Manually annotated by BRENDA team
gene treX, component of a treXYZ operon
-
-
Manually annotated by BRENDA team
Bacillus sp. CICIM 304
isolated by screening a soil sample collected from a hot spring of Tengchong volcano in Yunnan, China
UniProt
Manually annotated by BRENDA team
Bacillus sp. KSM-K16
KSM-K16
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
PY35
-
-
Manually annotated by BRENDA team
PY35
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
yeast, strain IGC 4051
-
-
Manually annotated by BRENDA team
cv. Nanicao
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
kidney bean
Swissprot
Manually annotated by BRENDA team
different isoforms Psisa1-Psisa3
-
-
Manually annotated by BRENDA team
strain JD210, isoamylase-hyperproducing mutant
-
-
Manually annotated by BRENDA team
strain MI-414, isomylase-hyperproducing mutant of SB-15
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
strain WU-5315, isoamylase hyperproducing mutant derived from JD-210
-
-
Manually annotated by BRENDA team
strain WU7211-2
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
alpha-D-glucan + H2O
?
show the reaction diagram
-
cleavage of alpha-D-glucosidic linkages of certain branched alpha-D-glucans
-
-
?
alpha-limit dextrin + H2O
linear maltooligosaccharides
show the reaction diagram
amylomaize + H2O
?
show the reaction diagram
-
debranching
-
-
?
amylopectin + H2O
?
show the reaction diagram
amylopectin + H2O
amylose + linear maltooligosaccharides
show the reaction diagram
amylopectin + H2O
linear maltooligosaccharides
show the reaction diagram
amylopectin + H2O
maltose + maltooligosaccharides
show the reaction diagram
amylopectin phi-dextrin + H2O
maltotetraose
show the reaction diagram
-
substrate on which all 3 types of debranching enzymes act
-
?
amylose + H2O
?
show the reaction diagram
-
short chain
-
-
?
amylose-extender amylopectin + H2O
maltose + maltooligosaccharides
show the reaction diagram
-
-
-
?
Arabidopsis starch + H2O
?
show the reaction diagram
beta limit dextrin + H2O
linear maltooligosaccharides
show the reaction diagram
-
-
-
?
beta limit dextrin + H2O
maltose + maltooligosaccharides
show the reaction diagram
from maize
-
-
?
beta-limit dextrin + H2O
?
show the reaction diagram
beta-limit dextrin + H2O
linear maltooligosaccharides
show the reaction diagram
beta-limit dextrin + H2O
maltose + ?
show the reaction diagram
-
-
-
-
?
branched chain oligosaccharides + H2O
maltotriose + higher maltosaccharides
show the reaction diagram
-
-
no detectable maltose released, characteristic action is hydrolysis of alpha 1,6-glucosidic linkages, but alpha 1,6 glucosidic linkages attaching maltose residues to chains of alpha 1,4-linked glucose residues are not readily hydrolyzed
?
corn amylopectin + H2O
?
show the reaction diagram
D-glucose + H2O
?
show the reaction diagram
-
59% of the activity with dextrin as substrate
-
-
?
dextrin + H2O
?
show the reaction diagram
-
enzyme activity 100%
-
-
?
di-O-alpha-maltosyl-beta-cyclodextrin + H2O
maltose + beta-cyclodextrin
show the reaction diagram
-
-
-
-
r
glycogen + H2O
?
show the reaction diagram
glycogen + H2O
linear maltooligosaccharides
show the reaction diagram
glycogen + H2O
maltodextrin
show the reaction diagram
glycogen + H2O
maltose + maltooligosaccharides
show the reaction diagram
maize beta-limit dextrin + H2O
?
show the reaction diagram
-
-
-
?
maize starch + H2O
?
show the reaction diagram
maltodextrin + H2O
?
show the reaction diagram
maltopentaose + H2O
?
show the reaction diagram
-
sugar immobilized on Sepharose 4B
-
-
?
maltose + H2O
?
show the reaction diagram
maltotriose + H2O
?
show the reaction diagram
-
activity 28%
-
-
?
oyster glycogen + H2O
?
show the reaction diagram
-
-
-
?
oyster gylcogen + H2O
maltose + maltooligosaccharides
show the reaction diagram
-
-
-
?
panose + H2O
D-glucose + maltose
show the reaction diagram
-
acts very slowly, small amounts of product formed after 24 h
-
?
phythoglycogen + H2O
?
show the reaction diagram
phytoglycogen + H2O
?
show the reaction diagram
phytoglycogen + H2O
maltose + maltooligosaccharides
show the reaction diagram
-
-
-
?
potato amylopectin + H2O
?
show the reaction diagram
-
-
-
?
potato starch, boiled + H2O
?
show the reaction diagram
pullulan + H2O
?
show the reaction diagram
pullulan + H2O
linear maltooligosaccharides
show the reaction diagram
-
-
-
?
saccharose + H2O
?
show the reaction diagram
-
activity 67%
-
-
?
soluble starch + H2O
?
show the reaction diagram
starch + H2O
?
show the reaction diagram
starch + H2O
linear maltooligosaccharides
show the reaction diagram
-
-
-
-
?
starch + H2O
maltose + maltooligosaccharides
show the reaction diagram
-
-
-
?
sticky rice starch + H2O
?
show the reaction diagram
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
amylopectin + H2O
?
show the reaction diagram
amylopectin + H2O
amylose + linear maltooligosaccharides
show the reaction diagram
Arabidopsis starch + H2O
?
show the reaction diagram
beta-limit dextrin + H2O
?
show the reaction diagram
glycogen + H2O
maltodextrin
show the reaction diagram
glycogen + H2O
maltose + maltooligosaccharides
show the reaction diagram
Q84UE6
-
-
-
?
maize starch + H2O
?
show the reaction diagram
-
-
-
-
?
starch + H2O
?
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
activates 10% at 0.5 mM, and 25% at 5 mM, Ca2+ highly stabilizes the enzyme at 80C
CaCl2
-
5 mM, 120% of activity
Co2+
activates 12% at 5 mM
Mn2+
activates 28% at 0.5 mM, and 41% at 5 mM
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(NH4)2Mo2O7
-
1 mM 22% inhibition, 10 mM 100% inhibition
-
2,4-Dinitrofluorobenzene
-
-
2-Hydroxy-5-nitrobenzyl bromide
-
-
ammonium sulfate
-
0.5 M, 30% residual activity
beta-cyclodextrin
-
29%, slightly inhibitory at 10 mM
cellobiose
-
10 mM, 0.4% amylose, relative activity 88%
corn amylose
-
-
-
cyclomaltoheptaose
-
10 mM, 0.4% amylose, relative activity 93%
D-glucono-delta-lactone
-
relative activity 87%
D-glucose
-
10 mM, 0.4% amylose, relative activity 95%
EDTA
-
5 mM, 80% residual activity
Fe3+
59% inhibition at 0.5 mM, 90% at 5 mM
guanidine hydrochloride
-
3 M, activity completely lost
Iodine
-
relative activity 93%
iodoacetate
isomaltose
-
10 mM, 0.4% amylose, relative activity 82%
maltose
-
10 mM, inhibition slightly, with 0.4% amylose relative activity 90%
maltotetraose
-
10 mM, 0.4% amylose, relative activity 50%
maltotriose
MgCl2
-
5 mM, 70% residual activity
MnCl2
-
5 mM, 80% residual activity
N-bromosuccinimide
NaF
-
1 mM, 19% inhibition
oligosaccharides with alpha-1,4-glucosidic linkages
-
-
p-chloromercuribenzoate
Phenylmercuriacetate
-
-
phenylmethanesulfonyl fluoride
9% inhibition at 5 mM
Sn2+
28% inhibition at 0.5 mM, 73% at 5 mM
sodium dodecylsulfate
-
1 mM, no residual activity
Succinic anhydride
-
relative activity 94%
xylose
-
10 mM, 0.4% amylose, relative activity 97%
Zn2+
64% inhibition at 0.5 mM, 88% at 5 mM
ZnCl2
-
5 mM, 20% residual activity
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
polyethyleneglycol 2000
-
maximal activation of 58% at 0.04% PEG 2000 Da
Triton X-100
-
reagent in the preincubation mixture at concentrations 0.05-1% activates the enzyme by 45-72%
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00012 - 0.00025
amylopectin
0.00002 - 0.00004
amylose-extender amylopectin
0.00018 - 0.00021
beta-limit dextrin
0.00062 - 0.00067
oyster glycogen
0.0054 - 0.0065
phytoglycogen
additional information
additional information
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.77
pullulan as substrate
0.95
pullulan as substrate; pullulan as substrate
3
-
pullulan, recombinant isoamylase
3.51
amylopectin as substrate
6.58
beta limit dextrin as substrate; beta limit dextrin as substrate
8.95
gylcogen as substrate; gylcogen as substrate
14.98
amylopectin as substrate; amylopectin as substrate
17.16
gylcogen as substrate
29.05
beta limit dextrin as substrate
51.2
-
rabbit liver glycogen
54.6
-
amylose, recombinant isoamylase
120
-
potato starch, recombinant isoamylase
154
-
rice starch, recombinant isoamylase
172
-
corn amylopectin, recombinant isoamylase
174
-
rabbit liver glycogen, recombinant isoamylase
182
-
oyster glycogen, recombinant isoamylase
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 6
-
amylopectin
6 - 7
-
-
6.3
-
only isoamylase with neutral pH optimum
6.5
broad optimum, profile overview
6.5 - 7
-
-
6.5 - 7.5
;
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1 - 5
-
starch, 0.1 M acetate-HCl buffer
2.2 - 7.8
-
0.1 M phosphate-citric acid buffer
4 - 8
-
not active below, amylopectin
5.5 - 9
over 90% of maximal activity within this range, profile overview
7.5 - 9.5
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 37
-
-
52
-
starch, 10 min reaction at pH 3.5
70
recombinant enzyme
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 60
-
not active above, starch
40 - 70
-
assay at
60
-
not active above, amylopectin
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50000
-
2 subunits, not linked covalently
75000
-
SU1
79000
-
maize gene su1 recombinant polypeptide, expressed in E. coli
80810
-
SB-15, DNA sequence analysis
86000 - 94000
88000
-
calculated by comparison with standard proteins
90000
-
undegraded polypeptide chain
95000
-
gel electrophoresis, low speed sedimentation
100000
gel filtration
120000
-
-
141000
-
isoamylase II, gel filtration
158000
-
ISA2 dimer, gel filtration and analytical ultracentrifugation
178000
-
native enzyme
180000
subunit ISA1, crystal structure analysis and SDS-PAGE
300000
wild type endosperm contains three high molecular mass ISA complexes resolved by gel filtration and native-PAGE. Two complexes of approximately 400 kDa contain both isoform ISA1 and isoform ISA2, and an approximately 300 kDa complex contains isoform ISA1 but not isoform ISA2
340000
-
native enzyme, gel filtration, TSKgel G3000SWxl
370000
gel permeation chromatography; gel permeation chromatography
400000
wild type endosperm contains three high molecular mass ISA complexes resolved by gel filtration and native-PAGE. Two complexes of approximately 400 kDa contain both isoform ISA1 and isoform ISA2, and an approximately 300 kDa complex contains isoform ISA1 but not isoform ISA2
420000
SDS-PAGE, homooligomer, five OsISA1
450000
native ISA1-ISA2 complex, gel filtration
490000
-
native enzyme, gel filtration, TSKgel G4000SWxl
510000
SDS-PAGE, heterooligomer, hetero-hexamer composed of five OsISA1 and one OsISA2; SDS-PAGE, heterooligomer, hetero-hexamer composed of five OsISA1 and one OsISA2
520000
-
gel filtration
750000
recombinant ISA1-ISA2 complex, gel filtration
830000
-
SDS-PAGE
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer or tetramer
-
the enzyme exists in two oligomeric states in solution, as a dimer and tetramer
homodimer
homooligomer
-
-
monomer
oligomer
tetramer
heterotetramer with PvISA1, 2 * 87000 PyISA1 and 2 * 93000 PvISA2, SDS-PAGE; heterotetramer with PvISA2, 2 * 87000 PvISA1 and 2 * 93000 PvISA2, SDS-PAGE
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified subunit ISA1 alone and in complex with maltoheptaose, X-ray diffraction structure determination at 2.3 A and 2.4 A, respectively; purified subunit ISA1 alone and in complex with maltoheptaose, X-ray diffrcation structure determination at 2.3 A and 2.4 A, respectively. The subunit structure includes highly conserved catalytic (beta/alpha)8 A-domain (aa 184-750), N-terminal beta-sandwich domain (aa 76-183), and C-terminal beta-sandwich domain (aa 751-875)
crystallized from ammonium sulfate solution, crystals belong to orthorhombic space group P212121
-
TreX in complex with an acarbose ligand, microbatch method under oil at 18C, dimeric crystal from 16% PEG 8000, 0.2 M NaCl, and 0.1 M CHES buffer, pH 9.5, tetrameric crystal form from 2.2 M ammonium phosphate and 0.1 M Tris-HCl buffer, pH 8.5. For the acarbose intermediate complex crystal, 0.1% acarbose is added to the protein, followed by incubation for 1 h prior to the setup of the crystal in 8% PEG 3000, 0.2M lithium sulfate, and 0.1 M imidazole buffer, pH 8.0, cyroprotection by 20% glycerol in mother liquor, in both crystal forms, the asymmetric unit consists of one dimer, X-ray diffraction structure determination and analysis at 2.8-3.0 A resolution
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.5 - 7.5
-
-
136806
3.5 - 5.5
-
-
136803
4.5 - 9
purified native enzyme, 50C, 1 h, over 70% activity remaining
732278
5 - 11.5
-
more than 50% of the original activity detectable between pH 6.6-10.5
136809
6.5 - 7
-
assayed at 22C enzyme displays broader activity and stability optima of pH 5.0 -7.5
136813
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
recombinant wild-type and His-tagged enzyme are more stable at room temperature than at 4C
10 - 40
-
temperature treatment does not influence the enzyme activity
21
-
recombinant wild-type and His-tagged enzyme are more stable at room temperature than at 4C
30 - 70
purified native enzyme, pH 6.0, 1 h, over 70% activity remaining
40
-
stable up to 40C, at temperature below 30C and above 65C enzyme is almost inactive
40 - 50
hetero-oligomer, loss of 50% activity; hetero-oligomer, loss of 50% activity
40
homooligomer, complete loss of activity
50
-
amylopectin, incubated in the absence of substrate enzyme stable only up to 45C
60
-
activity decreased about 95%
65
-
heating 10 min at 65C results in complete loss of activity
75
-
2 h, recombinant wild-type enzyme and His-tagged enzyme are sstable
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Pseudomonas isoamylase immobilized physically by raw starch adsorption results in an increase in pH and temperature stability and a retention of about 75% of activity even after 75 days
quite stable in maltose-containing buffer
-
very unstable, even at low temperatures
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80C, two weeks, 30% loss of activity
-
4C for 5 months 85% enzyme activity retained, preservation with 0.1% potassium sorbate isoamylase activity was not reduced until 2 months of incubation
-
4C several months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, DEAE cellulose column chromatography, CM-cellulose column chromatography, and DEAE-Sephadex gel filtration
-
copurification of native isozymes ISA1 and ISA2 partially from leaves by gel filtration, copurification of TAP-tagged ISA1 and ISA2 from Escherichia coli strain BL21(DE3) by tandem-affinity chromatography on TEV and calmodulin, followed by gel filtration
HiTrapQ column chromatography and Superdex 200 gel filtration
HitrapQ column chromatography and TSKgel DEAE-5PW gel filtration
-
native extracellular enzyme 369fold from culture supernatant by ultrafiltration, ammonium sulfate fractionation, gel filtration, two different steps of anion exchange chromatography, and dialysis, to homogeneity
native subunit ISA2 from leaves by anion exchange chromatography, ultrafiltration, and gel filtration, separation from subunit ISA1; native subunit ISAI from leaves by anion exchange chromatography, ultrafiltration, and gel filtration, separation from subunit ISA2
-
Q sepharose column chromatography; Q sepharose column chromatography; Q sepharose column chromatography
raw starch desorption adsorption
recombinant C-terminally His8-tagged single subunit ISA1 or ISA1 in complex with His8-tagged ISA2 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography; recombinant C-terminally His8-tagged single subunit ISA2 or His8-tagged ISA2 in complex with ISA1 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant His-tagged subunit ISA1 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant protein by His tag column chromatography
-
recombinant wild-type and His-tagged enzyme
-
recombinant wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography
-
wild type enzyme by HitrapQ HP and TSKgel G3000SWXL column chromatography, recombinant OsISA2 by HitrapQ HP column chromatography; wild type enzyme by HitrapQ HP and TSKgel G3000SWXL column chromatography, recombinant OsISA2 by HitrapQ HP column chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloned to an expression vector with a T7lac promoter. Both wild-type and His-tagged isoamylases are expressed in Escherichia coli
-
cloning of cDNAs of different isoforms, Psisa1-Psisa3, DNA and amino acid sequence determination and analysis, phylogenetic analysis and sequence comparisons, overview
-
coexpression of wild-type His-tagged ISA1 and ISA2, and of mutant D367A ISA1 with wild-type ISA2, in Escherichia coli strain BL21(DE3)
DNA and amino acid sequence determination and analysis, phylogenetic tree
DNA and amino acid sequence determination and analysis, sequence comparisons
-
expressed in Escherichia coli
-
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli BL21(DE3) Star cells
-
expressed in Saccharomyces cerevisiae
expression in Escherichia coli
-
expression of in Escherichia coli
-
expression of OsISA2 in Escherichia coli; expression of OsISA2in Escherichia coli
expression of wild-type and mutant enzymes in Escherichia coli, sequence comparison
-
gene ArDBE, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis
gene isa1, single expression of C-terminally His8-tagged subunit in Escherichia coli strain BL21(DE3), co-expression of untagged ISA1 subunit with C-terminally His8-tagged subunit ISA2 in Escherichia coli; gene isa2, single expression of C-terminally His8-tagged subunit in Escherichia coli strain BL21(DE3) or co-expression with untagged ISA1 subunit in Escherichia coli
gene isa2, single expression of C-terminally His8-tagged subunit in Escherichia coli strain BL21(DE3) or co-expression with untagged ISA1 subunit in Escherichia coli; gene su-1, single expression of C-terminally His8-tagged subunit ISA1 in Escherichia coli strain BL21(DE3), co-expression of untagged ISA1 subunit with C-terminally His8-tagged subunit ISA2 in Escherichia coli. Zea mays ISA1 subunit is heterologously expressed in Arabidopsis thaliana lacking endogenous ISA1 or lacking both native ISA1 and ISA2 via transfection with Agrobacterium tumefaciens strain GV3101. The maize protein functions in Arabidopsis leaves to support nearly normal starch metabolism in the absence of any native ISA1 or ISA2
-
gene su1, expressed in Escherichia coli, pAR4
-
gene treX, component of a treXYZ operon, DNA and amino acid sequence determination and analysis, sequence comparison, genetic organization in the treXYZ operon, overview
-
iam gene cloned and expressed in Escherichia coli JM101, pMON17481, production of a nonsecreted signal peptide-less isoamylase
-
into Agrobacterium tumefaciens strain GV310, GFP/GUS promoter fusion constructs for AtISA1, AtISA2, AtISA3 into Arabidopsis thaliana using the floral dip method
-
introduction of ISA1 gene from Triticum aestivum (TaISA1 from the D genome donor Aegilops tauschii) into a rice sugary-1 mutant line EM914 in which starch is completely replaced by phytoglycogen in the endosperm
-
isoamylase gene is a single copy in rice genome, located on chromsome 8
-
plasmid pIAM275, pUC9 carrying iam gene, Escherichia coli, recominant plasmid does not direct the synthesis of isoamylase in Escherichia coli , sequenced
-
recombinant GST-ISA2 fusion protein is expressed in Escherichia coli Rosetta DE3-pLysS cells
the STA7 locus encodes ISA1, recombinant expression of His-tagged subunit ISA1 in Escherichia coli strain BL21(DE3); the STA8 locus encodes ISA2, recombinant expression of HA-tagged ISA2 in strain BafV13 and BafO6
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
isoform ISA2 transcript is relatively abundant during periods of either starch biosynthesis or catabolism
the isoform ISA1 transcript level is elevated in tissues where starch is synthesized
the isoform ISA1 transcript level is low during starch degradation
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D367A
site-directed mutagenesis of the catalytic nucleophile Asp-367 in ISA1, the mutation abolishes the catalytic activity of the enzyme complex
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
degradation
-
thermophilic enzyme shows a potential to be used in industry to degrade the debranching points of starch at a high temperature
detergent
food industry
medicine
-
diagnosis of acute pancreatitis in humans
nutrition
synthesis
-
the enzyme is used for debranching of amylomaize in the production of cycloamylose, natural amylopectin containing starch with enhanced conversion yield after debranching, use of Thermus aquaticus 4-alpha-glucanotransferase for conversion of debranched amylomaize amylose and amylomaize amylopectin into cycloamylose, overview
additional information
Show AA Sequence (114 entries)
Please use the Sequence Search for a certain query.