Information on EC 3.2.1.116 - glucan 1,4-alpha-maltotriohydrolase

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

EC NUMBER
COMMENTARY hide
3.2.1.116
-
RECOMMENDED NAME
GeneOntology No.
glucan 1,4-alpha-maltotriohydrolase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of (1->4)-alpha-D-glucosidic linkages in amylaceous polysaccharides, to remove successive maltotriose residues from the non-reducing chain ends
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
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SYSTEMATIC NAME
IUBMB Comments
4-alpha-D-glucan maltotriohydrolase
Cf. EC 3.2.1.2 (beta-amylase), EC 3.2.1.60 (glucan 1,4-alpha-maltotetraohydrolase) and EC 3.2.1.98 (glucan 1,4-alpha-maltohexaosidase). The products have the alpha-configuration.
CAS REGISTRY NUMBER
COMMENTARY hide
91273-84-6
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain Ah-36
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Manually annotated by BRENDA team
strain Ah-36
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Manually annotated by BRENDA team
strain NTU22
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Manually annotated by BRENDA team
strain NTU22
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Manually annotated by BRENDA team
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
amylopectin + H2O
maltotriose
show the reaction diagram
amylose + H2O
maltotriose
show the reaction diagram
beta-cyclodextrin + H2O
maltotriose
show the reaction diagram
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main product
?
corn amylopectin + H2O
maltotriose + D-maltose + D-glucose
show the reaction diagram
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-
-
-
?
corn amylose + H2O
maltotriose + D-maltose + D-glucose
show the reaction diagram
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better substrate than corn starch or corn amylopectin
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-
?
corn starch + H2O
maltotriose + D-maltose
show the reaction diagram
-
-
-
-
?
gamma-cyclodextrin + H2O
?
show the reaction diagram
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-
-
-
?
maltoheptaose + H2O
maltotriose
show the reaction diagram
maltohexaose + H2O
maltotriose
show the reaction diagram
maltooligosaccharide + H2O
maltotriose
show the reaction diagram
maltopentaose + H2O
maltotriose + D-maltose
show the reaction diagram
maltotetraose + H2O
maltotriose + D-glucose
show the reaction diagram
maltotriose + H2O
maltose + D-glucose
show the reaction diagram
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slow hydrolysis
alpha-configuration
?
raw sago starch + H2O
maltotriose + ?
show the reaction diagram
short-chain amylose + H2O
maltotriose
show the reaction diagram
soluble starch + H2O
maltotriose + ?
show the reaction diagram
starch + H2O
maltotriose
show the reaction diagram
starch + H2O
maltotriose + D-maltose + D-glucose
show the reaction diagram
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?
additional information
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
CsCl
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can partially substitute for NaCl at higher concentrations
KCl
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can partially substitute for NaCl at higher concentrations
NaCl
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requirement, maximum activity at 2.5 M NaCl, no activity below 1 M NaCl
RbCl
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can partially substitute for NaCl at higher concentrations
additional information
not influenced by Mg2+, Ca2+, Cs2+, Li+, and Mn2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
alpha-cyclodextrin
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competitive inhibitor, 0.25%
beta-cyclodextrin
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competitive inhibitor, 0.025%
Ca2+
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1 mM, 33% inhibition
Co2+
53% residual activity at 10 mM
Cu2+
19% residual activity at 10 mM
EDTA
23% residual activity at 10 mM
Fe3+
19% residual activity at 10 mM
N-bromosuccinimide
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1 mM, 73% inhibition
SDS
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0.1%, 80% inhibition
Urea
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1 M, 74% inhibition
Zn2+
18% residual activity at 10 mM
ZnCl2
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1 mM, complete inhibition
additional information
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not inhibited by 1 mM iodoacetic acid, 1 mM phenylmethylsulfonyl fluoride
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
49
amylopectin
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pH 9, 50C, Km-value shown as glucose equivalents
19
amylose
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pH 9, 50C, DP, 17, Km-value shown as glucose equivalents
6.8
gamma-cyclodextrin
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pH 9, 50C, Km-value shown as glucose equivalents
42
glycogen
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pH 9, 50C, from oyster, Km-value shown as glucose equivalents
25
starch
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pH 9, 50C, soluble, Km-value shown as glucose equivalents
additional information
additional information
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Km-value for soluble starch is 0.88 mg/ml
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.63
alpha-cyclodextrin
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pH 9, 50C, 0.25%
0.41
beta-cyclodextrin
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pH 9, 50C, 0.025%
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
160
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pH 9, 50C
additional information
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.7
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in the presence of 2.5 M NaCl
9
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assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8.5
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about 35% activity at pH 5.0, about 90% activity at pH 6.0, about 97% activity at pH 7.0, about 65% activity at pH 8.0, about 33% activity at pH 8.5
5.8 - 8.5
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pH 5.8: about 90% of activity maximum, pH 8.5: about 30% of activity maximum
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
55
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pH 9, 30 min, in the presence of 2.5 M NaCl
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 55
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about 50% of activity maximum at 25C and 55C
30 - 65
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about 55% activity at 30C, about 75% activity at 40C, 100% activity at 50C, about 50% activity at 60C, about 30% activity at 65C
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40000
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gel filtration
60000
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gel filtration
76000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5
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30 min, 50C, 2.5 M NaCl, 0.5% soluble starch, 80% loss of activity
646648
4.2
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30 min, 50C, 2.5 M NaCl, 80% loss of activity
646648
5 - 9
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stable in the pH range
646647
5
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30 min, 50C, 2.5 M NaCl, 44% loss of activity
646648
6 - 10
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30 min, 50C, 2.5 M NaCl, 0.5% soluble starch, stable
646648
6 - 8.6
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30 min, 50C, 2.5 M NaCl, stable
646648
10.5
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30 min, 50C, 2.5 M NaCl, 30% loss of activity
646648
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 50
at 30C, the enzyme maintains approximately above 90% activity until 6 h and 80% activity until 12 h. At 40C, the enzyme maintains approximately above 90% activity until 2 h, 80% activity until 4 h, 70% activity until 8 h, and 60% activity until 12 h. At 50C, the enzyme maintains approximately 50% activity until 1 h, 30% activity until 6 h, and 20% activity until 12 h
40
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unstable above
50
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30 min, 2.5 M NaCl, pH 4.2: 80% loss of activity, pH 5.0: 44% loss of activity, pH 6.0-8.6: stable, pH 10.5: 30% loss of activity
55
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pH 8.0, 30 min, 2.5 M NaCl, 50% loss of activity, no loss of activity in the presence of 0.5% soluble starch
60
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3 h, about 70% of the activity remains
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
0.5% soluble starch stabilizes
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2.5 M NaCl stabilizes
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activity is irreversibly lost at low ionic strength
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no loss of activity after dialysis
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Acetone
benzene
93% activity at 20% (v/v)
chloroform
Ethanol
hexane
the enzyme exhibits extreme stability toward 20% (v/v) hexane (99% activity)
Methanol
n-Butanol
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2000fold, in the presence of 2.5 M NaCl
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His-tag affinity column chromatography
HiTrap Q column chromatography, and gel filtration
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
nutrition
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enzyme produced on the industrial scale for the production of high maltotriose-containing syrup