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Information on EC 3.2.1.68 - isoamylase and Organism(s) Pseudomonas amyloderamosa and UniProt Accession P10342

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EC Tree
     3 Hydrolases
         3.2 Glycosylases
             3.2.1 Glycosidases, i.e. enzymes that hydrolyse O- and S-glycosyl compounds
                3.2.1.68 isoamylase
IUBMB Comments
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.
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This record set is specific for:
Pseudomonas amyloderamosa
UNIPROT: P10342
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Word Map
The taxonomic range for the selected organisms is: Pseudomonas amyloderamosa
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
isoamylase, glycogen debranching enzyme, starch debranching enzyme, alpha-1,6-glucosidase, glycogen-debranching enzyme, isoamylase1, isoamylase 3, atisa3, isoamylase-type debranching enzyme, atisa1, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycogen-6-glucanohydrolase
-
bacterial isoamylase
-
-
debranching enzyme
-
-
-
-
glycogen 6-glucanohydrolase
-
-
glycogen alpha-1,6-glucanohydrolase
-
-
Pseudomonas isoamylase
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
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
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
alpha-limit dextrin + H2O
linear maltooligosaccharides
show the reaction diagram
-
-
-
?
amylopectin + H2O
maltose + maltooligosaccharides
show the reaction diagram
isoamylase is a direct debranching enzyme with the ability to cleave all the alpha-1,6 linkages of glycogen both inner and outer branching points of soluble amylopectin
-
-
?
beta-limit dextrin + H2O
linear maltooligosaccharides
show the reaction diagram
-
-
-
?
glycogen + H2O
maltose + maltooligosaccharides
show the reaction diagram
-
-
-
?
alpha-D-glucan + H2O
?
show the reaction diagram
-
cleavage of alpha-D-glucosidic linkages of certain branched alpha-D-glucans
-
-
?
amylopectin + H2O
?
show the reaction diagram
-
the enzyme can debranch almost all of component chains of amylopectin
-
-
?
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
-
?
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
-
-
-
-
?
di-O-alpha-maltosyl-beta-cyclodextrin + H2O
maltose + beta-cyclodextrin
show the reaction diagram
-
-
-
-
r
glycogen + H2O
maltose + maltooligosaccharides
show the reaction diagram
maltose + H2O
?
show the reaction diagram
phytoglycogen + H2O
?
show the reaction diagram
-
the enzyme can debranch almost all of component chains of phytoglycogen
-
-
?
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
additional information
?
-
-
isoamylase is one of the starch-debranching enzymes which catalyzes the hydrolysis of alpha-1,6-glucosidic linkages specific in alpha-glucans including amylopectin, branched starch, and glycogen
-
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,4-dinitrofluorobenzene
-
-
2-Hydroxy-5-nitrobenzyl bromide
-
-
cellobiose
-
10 mM, 0.4% amylose, relative activity 88%
corn amylose
-
-
-
Cu2+
-
relative activity 78%
CuCl2
-
1 mM, 30% inhibition
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%
guanidine hydrochloride
-
3 M, activity completely lost
HgCl2
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
N-bromosuccinimide
-
-
NaF
-
1 mM, 19% inhibition
oligosaccharides with alpha-1,4-glucosidic linkages
-
-
p-chloromercuribenzoate
Phenylmercuriacetate
-
-
SDS
-
0,1%, activity completely lost
Succinic anhydride
-
relative activity 94%
Urea
-
20% activity retained
xylose
-
10 mM, 0.4% amylose, relative activity 97%
additional information
-
inhibitory effect of pressure and agitation speed on isoamylase activity in supercritical CO2, overview
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
polyethyleneglycol 2000
-
maximal activation of 58% at 0.04% PEG 2000 Da
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 4
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
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
52
-
starch, 10 min reaction at pH 3.5
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 60
-
not active above, starch
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
ISOA_PSEAY
776
1
83627
Swiss-Prot
-
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
95000
x * 95000, SDS-PAGE
46000
-
2 * 46000, sedimentation equilibrium
50000
-
2 subunits, not linked covalently
52000
-
2 * 52000, gel filtration
80810
-
SB-15, DNA sequence analysis
86000 - 94000
88000
-
calculated by comparison with standard proteins
90000
-
undegraded polypeptide chain
94000
95000
-
gel electrophoresis, low speed sedimentation
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 95000, SDS-PAGE
dimer
monomer
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystallized from ammonium sulfate solution, crystals belong to orthorhombic space group P212121
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.5 - 7.5
-
-
136806
3.5 - 5.5
-
-
136803
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
-
not stable above
60
-
activity decreased about 95%
65
-
heating 10 min at 65°C results in complete loss of activity
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
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C for 5 months 85% enzyme activity retained, preservation with 0.1% potassium sorbate isoamylase activity was not reduced until 2 months of incubation
-
4°C several months
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
raw starch desorption adsorption
HitrapQ column chromatography and TSKgel DEAE-5PW column chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Saccharomyces cerevisiae
expressed in Escherichia coli BL21 cells
-
plasmid pIAM275, pUC9 carrying iam gene, Escherichia coli, recominant plasmid does not direct the synthesis of isoamylase in Escherichia coli , sequenced
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
food industry
saccharification potential of isoamylases is employed in food industry for preparation of high glucose syrup, maltose, maltitol, trehalose, cyclodextrin and resistant starch from starch
analysis
nutrition
additional information
isoamylase can potentially be used in the elucidation of fine structures of polysaccharides and related alpha-glucans and can be also be used as effective additive in dishwashing and laundry detergents
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Amemura, A.; Chakraborty, R.; Fujita, M.; Noumi, T.; Masamitsu, F.
Cloning and nucleotide sequence of the isoamylase gene from Pseudomonas amyloderamosa SB-15
J. Biol. Chem.
263
9271-9275
1988
Pseudomonas amyloderamosa
Manually annotated by BRENDA team
Amemura, A.; Konishi, Y.; Harada, T.
Molecular weight of the undegraded polypeptide chain of Pseudomonas amyloderamosa isoamylase
Biochim. Biophys. Acta
611
390-393
1980
Pseudomonas amyloderamosa
Manually annotated by BRENDA team
Kato, K.; Konishi, Y.; Amemura, A.; Harada, T.
Affinity chromatography of Pseudomonas isoamylase on cross-linked amylose gel
Agric. Biol. Chem.
41
2077-2080
1977
Pseudomonas amyloderamosa
-
Manually annotated by BRENDA team
Kitagawa, H.; Amemura, A.; Harada, T.
Studies on the inhibition and molecular properties of crystalline Pseudomonas isoamylase
Agric. Biol. Chem.
39
989-994
1975
Pseudomonas amyloderamosa
-
Manually annotated by BRENDA team
Lee, E.Y.C.; Whelan, W.J.
Glycogen and starch debranching enzymes
The Enzymes, 3rd Ed. (Boyer, P. D. , ed. )
5
191-234
1972
Cytophaga sp., Pseudomonas amyloderamosa
-
Manually annotated by BRENDA team
Yokobayashi, K.; Misaki, A.; Harada, T.
Purification and properties of Pseudomonas isoamylase
Biochim. Biophys. Acta
212
458-469
1970
Pseudomonas amyloderamosa
Manually annotated by BRENDA team
Fujita, F.; Sakai, S.; Futai, M.; Amemura, A.
Characterization of an isoamylase-hyperproducing mutant of Pseudomonas amyloderamosa
Agric. Biol. Chem.
54
2315-2321
1990
Pseudomonas amyloderamosa, Pseudomonas amyloderamosa MI-414
-
Manually annotated by BRENDA team
Katsuya, Y.; Mezaki, Y.; Kubota, M.; Matsuura, Y.
Three-dimensional structure of Pseudomonas isoamylase at 2.2 A resolution
J. Mol. Biol.
281
885-897
1998
Pseudomonas amyloderamosa
Manually annotated by BRENDA team
Ara, K.; Saeki, K.; Ito, S.
Purification and characterization of an alkaline isomylase from an alkalophilic strain of Bacillus
J. Gen. Microbiol.
139
781-786
1993
Bacillus sp. (in: Bacteria), Pseudomonas amyloderamosa, Pseudomonas amyloderamosa SMP1
-
Manually annotated by BRENDA team
Lin, L.L.; Fang, T.Y.; Chu, W.S.; Hsu, W.H.
Improved elution of isoamylase adsorbed on raw starch and the preservation of purified enzyme
Lett. Appl. Microbiol.
19
383-385
1994
Pseudomonas amyloderamosa, Pseudomonas amyloderamosa WU-5315
-
Manually annotated by BRENDA team
Krohn, M.; Barry, G.F.; Kishore, G.M.
An isoamylase with neutral pH optimum from a Flavobacterium species: cloning, characterization and expression of the iam gene
Mol. Gen. Genet.
254
469-478
1997
Flavobacterium sp., Pseudomonas amyloderamosa, Pseudomonas amyloderamosa JD210, Pseudomonas amyloderamosa SMP1
Manually annotated by BRENDA team
Yoon, S.; Robyt, J.F.
Activation and stabilization of 10 starch-degrading enzymes by Triton X-100, polyethylene glycols, and polyvinyl alcohols
Enzyme Microb. Technol.
37
556-562
2005
Pseudomonas amyloderamosa
-
Manually annotated by BRENDA team
Kang, H.K.; Cha, H.; Yang, T.J.; Park, J.T.; Lee, S.; Kim, Y.W.; Auh, J.H.; Okada, Y.; Kim, J.W.; Cha, J.; Kim, C.H.; Park, K.H.
Enzymatic synthesis of dimaltosyl-beta-cyclodextrin via a transglycosylation reaction using TreX, a Sulfolobus solfataricus P2 debranching enzyme
Biochem. Biophys. Res. Commun.
366
98-103
2008
Pseudomonas amyloderamosa
Manually annotated by BRENDA team
Park, I.M.; Ibanez, A.M.; Shoemaker, C.F.
Rice starch molecular size and its relationship with amylose content
Starch Staerke
59
69-77
2007
Pseudomonas amyloderamosa
-
Manually annotated by BRENDA team
Wang, S.; Lai, J.; Huang, M.; Tai, C.; Liu, H.
Deactivation of isoamylase and β-amylase in the agitated reactor under supercritical carbon dioxide
Bioprocess Biosyst. Eng.
33
1007-1015
2010
Pseudomonas amyloderamosa, Pseudomonas amyloderamosa WU7211-2
Manually annotated by BRENDA team
Ray, R.
Microbial isoamylases: An overview
Am. J. Food Technol.
6
1-18
2011
Myroides odoratus, Pseudomonas amyloderamosa (P10342), Pseudomonas amyloderamosa SB-15 (P10342)
-
Manually annotated by BRENDA team
Kobayashi, T.; Sasaki, S.; Utsumi, Y.; Fujita, N.; Umeda, K.; Sawada, T.; Kubo, A.; Abe, J.; Colleoni, C.; Ball, S.; Nakamura, Y.
Comparison of chain-length preferences and glucan specificities of isoamylase-type alpha-glucan debranching enzymes from rice, Cyanobacteria, and Bacteria
PLoS ONE
11
e0157020
2016
Crocosphaera subtropica ATCC 51142, Escherichia coli, Oryza sativa, Pseudomonas amyloderamosa, Synechococcus elongatus, Synechococcus elongatus PCC7942
Manually annotated by BRENDA team