Information on EC 3.2.1.41 - pullulanase

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

EC NUMBER
COMMENTARY hide
3.2.1.41
-
RECOMMENDED NAME
GeneOntology No.
pullulanase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of (1->6)-alpha-D-glucosidic linkages in pullulan, amylopectin and glycogen, and in the alpha- and beta-limit dextrins of amylopectin and glycogen
show the reaction diagram
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-
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
hydrolysis of O-glycosyl bond
additional information
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
starch degradation
-
-
SYSTEMATIC NAME
IUBMB Comments
pullulan 6-alpha-glucanohydrolase
Different from EC 3.2.1.142 (limit dextrinase) in its action on glycogen, and its rate of hydrolysis of limit dextrins. Its action on amylopectin is complete. Maltose is the smallest sugar that it can release from an alpha-(1->6)-linkage.
CAS REGISTRY NUMBER
COMMENTARY hide
9075-68-7
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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EPZ37738
GenBank
Manually annotated by BRENDA team
-
EPZ37738
GenBank
Manually annotated by BRENDA team
oat
-
-
Manually annotated by BRENDA team
strain FDTA 13 isolated from tapioca soil and mutant NTG04-B4
-
-
Manually annotated by BRENDA team
strain AN-7 from soil in India
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-
Manually annotated by BRENDA team
sugar beet
-
-
Manually annotated by BRENDA team
previously B. thermoleovorans US105
-
-
Manually annotated by BRENDA team
previously B. thermoleovorans US105
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
L. cv. Chariot
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain PCC73102
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-
Manually annotated by BRENDA team
strain PCC73102
-
-
Manually annotated by BRENDA team
optimized growth conditions: medium with 1.5 g per l maltose and 8.3 g per l yeast extract
-
-
Manually annotated by BRENDA team
spinach
-
-
Manually annotated by BRENDA team
mutant strains
-
-
Manually annotated by BRENDA team
group B Streptococcus type I
-
-
Manually annotated by BRENDA team
Thermoanaerobium sp.
-
-
-
Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
type I enzyme
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
HB8
-
-
Manually annotated by BRENDA team
additional information
Promozyme 200 L purchased from Novo Nordisk, Denmark
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-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3 glycogen + H2O
4 maltotriose
show the reaction diagram
4-nitrophenyl alpha-D-maltoheptaoside + H2O
4-nitrophenol + alpha-D-maltoheptaose
show the reaction diagram
4-nitrophenyl alpha-D-maltotrioside + H2O
4-nitrophenol + alpha-D-maltotriose
show the reaction diagram
42-O-beta-D-galactosyl-maltose + beta-cyclodextrin
(Gal-G2)-beta-cyclodextrin + (Gal-G2)2-beta-cyclodextrin
show the reaction diagram
-
analysis of reaction conditions and structural identification of products
-
-
r
6-O-alpha-maltosyl cyclodextrin + H2O
glucose + cyclodextrin
show the reaction diagram
6-O-alpha-maltotriosyl cyclomaltoheptaose + H2O
glucose + cyclodextrin
show the reaction diagram
6-O-alpha-maltotriosyl cyclomaltohexaose + H2O
glucose + cyclodextrin
show the reaction diagram
amylopectin + H2O
?
show the reaction diagram
amylopectin + H2O
D-glucose + maltose + maltotriose
show the reaction diagram
amylopectin + H2O
malto-2-4-oligosaccharide + ?
show the reaction diagram
-
-
-
-
?
amylopectin + H2O
maltooligosaccharides + maltose
show the reaction diagram
amylopectin + H2O
maltotriose
show the reaction diagram
-
-
-
?
amylopectin + H2O
maltotriose + maltose + ?
show the reaction diagram
amylopectin + H2O
maltotriose + maltose + maltooligosaccharides
show the reaction diagram
amylopectin beta-limit dextrin + H2O
maltotriose + maltose
show the reaction diagram
amylose + H2O
?
show the reaction diagram
amylose + H2O
malto-2-4-oligosaccharide + ?
show the reaction diagram
-
-
-
-
?
amylose + H2O
maltooligosaccharides + maltose
show the reaction diagram
amylose + H2O
maltose + maltotriose
show the reaction diagram
amylose + H2O
maltotriose + maltose
show the reaction diagram
beta-cyclodextrin + H2O
?
show the reaction diagram
21% of activity compared to pullulan
-
-
?
beta-limit dextrin + H2O
?
show the reaction diagram
beta-limited dextrin + H2O
maltotriose + maltose + maltooligosaccharides
show the reaction diagram
cyclomaltooctaose + H2O
6,6-di-O-alpha-maltosyl-cG8
show the reaction diagram
-
gamma-cyclodextrin
-
-
?
dextran + H2O
?
show the reaction diagram
-
-
-
-
?
dextrin + H2O
maltooligosaccharides + maltose
show the reaction diagram
gamma-cyclodextrin + H2O
?
show the reaction diagram
71% of activity compared to pullulan
-
-
?
glycogen + H2O
?
show the reaction diagram
glycogen + H2O
malto-2-4-oligosaccharide + ?
show the reaction diagram
-
-
-
-
?
glycogen + H2O
maltooligosaccharides + maltose
show the reaction diagram
glycogen + H2O
maltotriose + maltose
show the reaction diagram
-
-
-
?
glycogen + H2O
maltotriose + maltose + maltooligosaccharides
show the reaction diagram
glycogen beta-limit dextrin + H2O
maltotriose + maltose + maltosetetraose
show the reaction diagram
guar galactomannan + H2O
?
show the reaction diagram
-
-
-
-
?
maltoheptaose + H2O
maltotriose + maltose + D-glucose
show the reaction diagram
maltohexaose + H2O
maltotriose + maltose + D-glucose + maltotetraose
show the reaction diagram
maltopentaose + H2O
maltotriose + maltose
show the reaction diagram
maltose + cG8
6-O-alpha-maltosyl-cG8
show the reaction diagram
-
through the reversed action
-
-
r
maltotetraose + H2O
maltose + glucose + maltotriose
show the reaction diagram
phytoglycogen + H2O
?
show the reaction diagram
9% of activity compared to pullulan
-
-
?
potato starch + H2O
?
show the reaction diagram
pullulan + H2O
?
show the reaction diagram
pullulan + H2O
malto-2-4-oligosaccharide + ?
show the reaction diagram
-
best substrate
-
-
?
pullulan + H2O
maltotriose
show the reaction diagram
pullulan + H2O
maltotriose + ?
show the reaction diagram
pullulan + H2O
maltotriose + maltose
show the reaction diagram
rice flour + H2O
?
show the reaction diagram
soluble starch
?
show the reaction diagram
-
-
-
?
soluble starch
maltotriose + maltose
show the reaction diagram
soluble starch + H2O
?
show the reaction diagram
soluble starch + H2O
malto-2-4-oligosaccharide + ?
show the reaction diagram
-
-
-
-
?
soluble starch + H2O
maltotriose + maltose
show the reaction diagram
soluble starch + H2O
maltotriose + maltose + maltooligosaccharides
show the reaction diagram
12.8% of the activity with pullulan
-
-
?
starch + H2O
?
show the reaction diagram
starch + H2O
D-glucose + maltose + maltotriose
show the reaction diagram
starch + H2O
maltose + maltotriose
show the reaction diagram
starch + H2O
maltotriose
show the reaction diagram
metroxylon sagu + H2O
debranched starch
show the reaction diagram
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
maltooligosaccharides + maltose
show the reaction diagram
pullulan + H2O
maltotriose
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cd2+
-
highly stimulated
Hg2+
-
complete inactivating
NaCl
-
the enzyme is active over a wide range of concentrations (04.5 M) of NaCl
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
2-mercaptoethanol
5 mM, 51% residual activity
alpha-cyclodextrin
ammonium sulfate
0.5 mM, 36% residual activity
beta-cyclodextrin
CuCl2
5 mM, no residual activity
cyclodextrin
-
Cyclodextrins
-
cyclomaltoheptaose
-
-
cyclomaltohexaose
-
-
EGTA
-
1-10 mM, 65-70% inhibition
gamma-cyclodextrin
-
10 mM, 20% loss of activity
Guanidine HCl
iodoacetamide
N-bromosuccinimide
NH4+
EPZ37738
2 mM, 49% of initial activtiy
p-chloromercuribenzoate
phenylmethylsulfonyl fluoride
-
partially
Pullulan
-
substrate inhibition above 0.1%
Sodium dodecyl sulfate
sodium dodecylsulfate
1 mM, no residual activity
Triton X-100
ZnCl2
5 mM, 51% residual activity
additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
bovine serum albumin
-
assists reactivation after unfolding
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Brij-35
-
stimulates at 0.025-0.10% v/v
dithiothreitol
glutathione-SH
-
1-10 mM, increase of activity by 100%
Guanidine HCl
0.1 mM, 1.1fold activation
maltodextrin
-
activation is caused by an endotransglycosylation reaction catalyzed by EC 3.2.1.41
mercaptoethanol
10 mM, 1.1fold asctivation
thioredoxin
upon preincubation with both dithiothreitol and thioredoxin, the initial reaction velocity increases by a factor of two
Triton X-100
Tween 80
-
stimulates at 0.025-0.10% v/v
Tween-20
EPZ37738
1% v/v, 143% of initial activtiy
Urea
1.0-5.0 mM, 1.1fold activation
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.037 - 0.664
4-nitrophenyl alpha-D-maltotrioside
4.1
beta-limit dextrin
pH 6.0, 37C
0.021 - 2.5
Pullulan
additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.18
amylopectin
Oryza sativa
-
37C, pH 6.0
3.31
beta-limit dextrin
Oryza sativa
-
37C, pH 6.0
23.1 - 4830
Pullulan
2.89 - 877.9
soluble starch
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.2
-
substrate guar galactomannan, pH 4.5, 37C
3.2
-
of the periplasmic homogenate before purification
12
-
in the supernatant
15.2
-
substrate pullulan, pH 4.5, 37C
15.5
substrate phytoglycogen, pH 6.0, 37C
16.5 - 29.8
32.7
substrate amylopectin, pH 6.0, 37C
36
-
of the periplasmic homogenate after purification
151.5
-
after purification
165.9
substrate pullulan, pH 6.0, 37C
223.9
substrate beta-limit dextrin, pH 6.0, 37C
1352 - 1790
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 3.5
amylolytic activity towards starch
5
optimum pH for pullulanase activity
5
optimum pH for amylase activity
5.5 - 6.5
-
maltose + cyclomaltooctaose
7.5
-
pullulytic activity
8.5 - 9
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2 - 5
pH 2.0: about 60% of maximal activity, pH 5.0: about 60% of maximal activity, , amylolytic activity toward starch
4 - 12
-
-
4
-
not active below, maltose + cyclomaltooctaose
4 - 7.5
4.3 - 7.2
pH 4.3: about 80% of maximal activity, pH 7.2: about 75% of maximal activity
4.5 - 6.5
-
over 89% of maximal activity between pH 5.0 and pH 6.5, over 75% at pH 4.5
4.5 - 7.8
5 - 10
-
-
5 - 7
-
-
5 - 11
more than 60% of maximum activity
5.5 - 8.5
-
pH 5.5: about 45% of maximal activity, pH 8.5: about 45% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
assay at; assay at
60 - 65
95 - 105
amylolytic activity toward starch
98
-
assay at
102 - 108
-
-
105
with 0.5 mM Ca2+
125
-
substrate: pullulan or starch, 5 mM Ca2+