Any feedback?
Information on EC 3.2.1.3 - glucan 1,4-alpha-glucosidase and Organism(s) Aspergillus niger and UniProt Accession P69328
for references in articles please use BRENDA:EC3.2.1.3Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
3.2.1.3 glucan 1,4-alpha-glucosidaseIUBMB Comments
Most forms of the enzyme can rapidly hydrolyse 1,6-alpha-D-glucosidic bonds when the next bond in the sequence is 1,4, and some preparations of this enzyme hydrolyse 1,6- and 1,3-alpha-D-glucosidic bonds in other polysaccharides. This entry covers all such enzymes acting on polysaccharides more rapidly than on oligosaccharides. EC 3.2.1.20 alpha-glucosidase, from mammalian intestine, can catalyse similar reactions.
Specify your search results
Word Map
- 3.2.1.3
- aspergillus
- niger
- glycogen
- alpha-amylase
- maltose
- corn
- awamori
-
amylolytic
- pompe
- sucrase
- myopathy
-
saccharification
- rhizopus
- lactase
-
brush
- sucrase-isomaltase
- acarbose
- dextrin
- glycogenosis
- maltotriose
- amylopectin
- flour
- amylases
-
starch-binding
- isomaltase
-
infantile
- maltodextrins
- occidentalis
- bran
- disaccharidase
- food industry
- pullulanase
-
liquefaction
- beta-cyclodextrins
-
debranching
- beta-amylase
-
carbohydrases
-
3.2.1.20
- cassava
- maltooligosaccharides
- trehalase
-
saccharify
-
bioethanol
- medicine
- industry
- energy production
- paper production
-
glucanotransferase
- nutrition
- maltoheptaose
- analysis
- maltotetraose
- biotechnology
- cgtase
- biofuel production
- isoamylase
-
liquefied
- 1-deoxynojirimycin
-
starchy
- synthesis
The taxonomic range for the selected organisms is: Aspergillus niger
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
Synonyms
glucoamylase, amyloglucosidase, acid maltase, maltase-glucoamylase, lysosomal alpha-glucosidase, maltase glucoamylase, gamma-amylase, glucose amylase, gam-1, glucoamylase p, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
1,4-alpha-D-glucan glucohydrolase
-
-
-
-
acid maltase
-
-
-
-
alpha-1,4-glucan glucohydrolase
amyloglucosidase
exo-1,4-alpha-glucosidase
-
-
-
-
GAI
-
-
-
-
GAII
-
-
-
-
gamma-amylase
Glucan 1,4-alpha-glucosidase
-
-
-
-
glucoamylase
glucose amylase
-
-
-
-
lysosomal alpha-glucosidase
-
-
-
-
Meiotic expression upregulated protein 17
-
-
-
-
alpha-1,4-glucan glucohydrolase
-
-
-
-
amyloglucosidase
-
-
-
-
gamma-amylase
-
-
-
-
glucoamylase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of terminal (1->4)-linked alpha-D-glucose residues successively from non-reducing ends of the chains with release of beta-D-glucose
the enzyme catalyses the cleavage of alpha-1,4- and alpha-1,6-glycosidic bonds. Glucoamylases use a classical acid/base Koshland-type inverting mechanism, releasing alpha-glucose from the nonreducing end of an alpha-glucan chain
Hydrolysis of terminal (1->4)-linked alpha-D-glucose residues successively from non-reducing ends of the chains with release of beta-D-glucose
cleavage of alpha-1,4-linkages is preferred to cleavage of alpha-1,6-linkages
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SYSTEMATIC NAME
IUBMB Comments
4-alpha-D-glucan glucohydrolase
Most forms of the enzyme can rapidly hydrolyse 1,6-alpha-D-glucosidic bonds when the next bond in the sequence is 1,4, and some preparations of this enzyme hydrolyse 1,6- and 1,3-alpha-D-glucosidic bonds in other polysaccharides. This entry covers all such enzymes acting on polysaccharides more rapidly than on oligosaccharides. EC 3.2.1.20 alpha-glucosidase, from mammalian intestine, can catalyse similar reactions.
CAS REGISTRY NUMBER
COMMENTARY
9032-08-0
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
4-nitrophenyl alpha-D-glucopyranoside + H2O
D-glucose + 4-nitrophenol
-
-
-
-
?
maltodextrin + H2O
maltodextrin + beta-D-glucose
-
during incubation 5-(hydroxymethyl)-2-furfuraldehyde is released, resulting in a glycated enzyme
-
-
?
maltose + H2O
beta-D-glucose + D-glucose
-
-
-
-
?
p-nitrophenyl-alpha-D-glucopyranoside + H2O
p-nitrophenol + D-glucose
-
-
-
-
?
glycogen + H2O
glucose + ?
-
shellfish glycogen, sweet-corn phytoglycogen, skate liver glycogen, human muscle glycogen, rabbit muscle glycogen, cat liver glycogen, cat liver glycogen. Incomplete conversion to glucose in absence of alpha-amylase
-
-
?
maltotriose + H2O
maltose + glucose
-
68% of the activity with starch
-
-
?
starch + H2O
beta-D-glucose + ?
-
-
-
-
?
starch + H2O
beta-D-glucose + ?
-
the starch binding domain of glucoamylase plays an active role in hydrolyzing raw starch and supports the enzyme adsorption to the cell wall where local increase of enzyme concentration may result in enhanced glucose flow to the cell
-
-
?
starch + H2O
beta-D-glucose + ?
-
the enzyme hydrolyzes alpha-1,4 glycosidic linkages in raw, sparsely soluble or soluble starches and related oligosaccharides with the inversion of the anomeric configuration to produce beta-glucose
-
-
?
starch + H2O
glucose + ?
-
with concentrated solutions of D-glucose, 10% w/v and 30% w/v, small amounts of isomaltose are produced as the sole reversion product
-
-
?
starch + H2O
glucose + ?
-
waxy-maize starch, waxy-sorghum starch, floridean starch. Incomplete conversion to glucose in absence of alpha-amylase
-
-
?
starch + H2O
starch + beta-D-glucose
-
hydrolysis of terminal 1,4-linked alpha-D-glucose residues successively from non-reducing ends of the starch chain, substrates corn cobs, maize starch, soluble starch, and wheat bran
-
-
?
starch + H2O
starch + beta-D-glucose
-
the enzyme is required for degradation of raw starch
-
-
?
starch + H2O
starch + beta-D-glucose
-
hydrolysis of solid-state starch from raw chestnut homogenate, composition, 30% of the raw chestnut is starch, overview
-
-
?
starch + H2O
starch + beta-D-glucose
-
the enzyme contains 7 subsites for substrate binding, subsite affinities
-
-
?
additional information
?
-
-
regulation mechanisms of enzyme expression, overview
-
-
?
additional information
?
-
-
immobilization of the enzyme on polyaniline polymer results in improved catalytic performance with decreased temperature optimum, and increased thermal stability and catalytic efficiency, overview
-
-
?
additional information
?
-
-
modelling of potato starch saccharification by Aspergillus niger, influences of assay conditions on activity, overview
-
-
?
additional information
?
-
-
glucoamylase functions via transient dimer formation during hydrolysis of insoluble substrates and address the question of the cooperative effect of starch binding and hydrolysis
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
starch + H2O
beta-D-glucose + ?
-
the starch binding domain of glucoamylase plays an active role in hydrolyzing raw starch and supports the enzyme adsorption to the cell wall where local increase of enzyme concentration may result in enhanced glucose flow to the cell
-
-
?
starch + H2O
starch + beta-D-glucose
-
hydrolysis of terminal 1,4-linked alpha-D-glucose residues successively from non-reducing ends of the starch chain, substrates corn cobs, maize starch, soluble starch, and wheat bran
-
-
?
starch + H2O
starch + beta-D-glucose
-
the enzyme is required for degradation of raw starch
-
-
?
additional information
?
-
-
regulation mechanisms of enzyme expression, overview
-
-
?
additional information
?
-
-
glucoamylase functions via transient dimer formation during hydrolysis of insoluble substrates and address the question of the cooperative effect of starch binding and hydrolysis
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ba2+
Sn2+
Zn2+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
methyl alpha-D-glucoside
-
competitive with maltose and non-competitive with starch
additional information
-
glucose, cycloheximide, and actinomycin D nearly completely suppresses enzyme expression, repression mechanism, overview
-
additional information
-
the deglycosylated enzyme is more sensitive to proteolytic degradation by subtilisin than the native enzyme
-
additional information
-
binding of a short heterobidentate inhibitor simultaneously directed toward the catalytic and starch binding domains causes dimerization of glucoamylase and not, an intramolecular conformational rearrangement mediated by linker flexibility
-
additional information
-
not inhibitory at 5 mM: Ag2+, Ca2+, Zn2+, Mg2+ and Cd2+ and EDTA
-
additional information
-
tannins isolated from extracts of pomegranate, cranberry, grape, and cocoa inhibit the activity of glucoamylase and alpha-amylase in vitro. In general, larger and more complex tannins, such as those in pomegranate and cranberry, more effectively inhibit the enzymes than less polymerized cocoa tannins
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Schardinger dextrin
-
slight stimulation of activity with maltose as substrate
sorbitol
-
presence of sorbitol and trehalose significantly increase the substrate affinity and catalytic efficiency of glucoamylases GAM-1 and GAM-2
trehalose
-
presence of sorbitol and trehalose significantly increase the substrate affinity and catalytic efficiency of glucoamylases GAM-1 and GAM-2
additional information
-
wheat bran, rice bran, and cellulose highly induce enzyme expression, maltose, cello-dextrins, cellulose, and cellulose- and hemicellulose-containing substrates also induce the enzyme to a lesser extent, induction mechanism, overview
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.2
maltotriose
-
15°C, pH 4.2, glucoamylase immobilized on macroporous silica
0.89
maltotriose
-
30°C, pH 4.2, glucoamylase immobilized on macroporous silica
0.017
starch
-
30°C, pH 4.2, glucoamylase immobilized on macroporous silica
additional information
additional information
-
kinetics and thermodynamics of wild-type and mutant enzymes
-
additional information
additional information
-
kinetics and thermodynamics, substrate specificity for fermentation
-
additional information
additional information
-
modelling of potato starch saccharification by commercial Aspergillus niger enzyme, kinetics
-
additional information
additional information
-
kinetics of starch hydrolysis by AMG modeled for two different systems, free enzyme in aqueous solution and entrapped enzyme within vesicles in aqueous suspension, overview, Michaelis-Menten kinetic model with product inhibition for intrinsic kinetics
-
additional information
additional information
-
thermodynamics and kinetics of starch hydrolysis, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.057 - 0.65
maltose
-
pH 4.5, 35°C, recombinant mutant D20C/A27C/S30P/G137A
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4.2
4.5
additional information
-
optimal pH for growth and enzyme production is pH 6.5
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
1.5 - 6.5
-
pH 1.5: about 70% of maximal activity with starch as substrate, about 60% of the activity with maltose as substrate, pH 6.5: about 15% of maximal activity with maltose as substrate, about 55% of maximal activity with starch as substrate
2 - 7
2 - 8
-
20% of maximal activity at pH 2.0 and 60% at pH 8.0, immobilized enzyme
2.5 - 8
-
pH 2.5: about 35% of maximal activity, immobilized enzyme, pH 2.5: about 55% of maximal activity, free enzyme, pH 8.0: about 25% of maximal activity, free enzyme, pH 8.0: about 50% of maximal activity, immobilized enzyme
2.5 - 8.5
-
soluble glucoamylase, approx. 70% of maximal activity at pH 2.5, approx. 50% of maximal activity at pH 8.5, crosslinked glucoamylase crystals, approx. 20% of maximal activity at pH 2.5, approx. 25% of maximal activity at pH 8.5
2 - 7
-
pH 2.0: about 55% of maximal activity, pH 7.0: about 25% of maximal activity, free and immobilized enzyme
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
40
50
55
60
additional information
-
optimal temperature for growth and enzyme production is 30°C
50
-
immobilized enzyme, the enzyme immobilized on polyaniline polymer shows a 10°C decreased temperature optimum
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30 - 60
-
30°C: about 30% of maximal activity, immobilized enzyme, 30°C: about 50% of maximal activity, free enzyme, 60°C: about 50% of maximal activity, free enzyme, 60°C: about 90% of maximal activity, immobilized enzyme
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
additional information
additional information
-
culture conditions for enzyme production, overview
additional information
-
optimization of growth conditions, enzyme expression depends on carbon source for growth, glucose nearly completely suppresses enzyme expression, while wheat bran, rice bran, and cellulose induce it, the enzyme prefers monosaccharides to disaccharides for growth, overview
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
the enzyme is a member of the glycoside hydrolase family 15 (GH15)
metabolism
dynamic metabolic response of Aspergillus niger to glucose perturbation, regulatory mechanism for reduced glucoamylase production. Reduction of the total adenine nucleotides and major precursor amino acids indicate the upregulated RNA synthesis is required to produce stress proteins, and partially explains the drop of glucoamylase production when Aspergillus niger experiences a fluctuated glucose concentration environment, adenine nucleotides dynamic concentration profiles, overview
physiological function
additional information
the relative orientation between the carbohydrate-binding domain (CBM) and the catalytic domain is flexible, as the domains can adopt different orientations independently of ligand binding, suggesting a role as an anchor to increase the contact time and the relative concentration of substrate near the active site. The C-terminal CBM adopts the well known beta-sandwich motif, which is a hallmark of carbohydrate-binding modules
physiological function
glucoamylase is an exoglucohydrolase that primarily catalyzes the hydrolysis of alpha-1,4 glycosidic linkages in raw starch and soluble oligosaccharides to generate beta-D-glucose
physiological function
reduction of the total adenine nucleotides and major precursor amino acids indicate the upregulated RNA synthesis is required to produce stress proteins, and partially explains the drop of glucoamylase production when Aspergillus niger experiences a fluctuated glucose concentration environment
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). AMYG_ASPNG
640
0
68309
Swiss-Prot
Secretory Pathway (Reliability: 2)
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
62000
68000
-
experimental molecular weight for glucoamylase 2, excellent agreement with the theoretical value
70000
-
calculated molecular weight for low-glycosylated linker variant dgGA
72063
-
1 * 93000, wild-type and mutant enzyme, SDS-PAGE, 1 * 72876, wild-type enzyme, mass spectrometry, 1 * 72063, mutant enzyme, mass spectrometry
72876
-
1 * 93000, wild-type and mutant enzyme, SDS-PAGE, 1 * 72876, wild-type enzyme, mass spectrometry, 1 * 72063, mutant enzyme, mass spectrometry
73000
90000
93000
-
1 * 93000, wild-type and mutant enzyme, SDS-PAGE, 1 * 72876, wild-type enzyme, mass spectrometry, 1 * 72063, mutant enzyme, mass spectrometry
62000
-
experimental molecular weight for low-glycosylated linker variant dgGA
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
-
complex formation with a heterobidentate ligand induces dimerization
monomer
additional information
monomer
-
a starch binding and a catalytic domain interspersed by a highly glycosylated polypeptide linker
monomer
-
1 * 93000, wild-type and mutant enzyme, SDS-PAGE, 1 * 72876, wild-type enzyme, mass spectrometry, 1 * 72063, mutant enzyme, mass spectrometry
additional information
the G1 isoform consists of a catalytic domain and a starch-binding domain connected by a heavily glycosylated linker region
additional information
-
the G1 isoform consists of a catalytic domain and a starch-binding domain connected by a heavily glycosylated linker region
additional information
glucoamylases consist of a catalytic domain and a carbohydrate-binding domain (CBM), with the latter being important for the interaction with the polymeric substrate. The relative orientation between the carbohydrate-binding domain (CBM) and the catalytic domain is flexible, as the domains can adopt different orientations independently of ligand binding, suggesting a role as an anchor to increase the contact time and the relative concentration of substrate near the active site. The C-terminal CBM adopts the well known beta-sandwich motif, which is a hallmark of carbohydrate-binding modules
additional information
the starch-binding domain (SBD) of glucoamylase from Aspergillus niger is a small globular protein containing a disulfide bond. The structure of the SBD has been determined by NMR. Cys509 and Cys604 are at the N- and C-termini, respectively, and they are linked by a disulfide bond, analysis of the conformation surrounding the disulfide bond, overview
additional information
-
the starch-binding domain (SBD) of glucoamylase from Aspergillus niger is a small globular protein containing a disulfide bond. The structure of the SBD has been determined by NMR. Cys509 and Cys604 are at the N- and C-termini, respectively, and they are linked by a disulfide bond, analysis of the conformation surrounding the disulfide bond, overview
additional information
-
the enzyme consists of a catalytic domain and a starch binding domain connected by an O-glycosylated peptide linker located at the N-terminus, the enzyme contains 7 subsites for substrate binding
additional information
-
fungal glucoamylases contain up to 7 subsites for substrate binding with highly varying affinity, the enzymes have two domains, namely a catalytic domain and a starch binding domain, the two domains are connected by an O-glycosylated polypeptide linker located at the N-terminus
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
glycoprotein
glycoprotein
proteolytic modification
glycoprotein
the linker region between starch-binding and catalytic domains is heavily glycosylated. The O-glycosylated residues are Ser467(443), Ser468(444), Thr476(452), Ser477(453), Ser483(459), Ser484(460) and Thr486(462)
glycoprotein
O-glycosylation is observed in enzyme AnGA, in particular in the linker domain, which connects to the carbohydrate-binding domain. Only two sites, at Ser483 and Ser484, can be modelled with confidence. The catalytic domain of AnGA has two resolved N-glycosylation sites at Asn195 and Asn419, with two GlcNAc residues visible at Asn195 and one at Asn419
glycoprotein
-
carbohydrate groups seem to play an important role in stabilizing the structure against inactivation by heat and storage
glycoprotein
-
effects of deglycosylation on enzyme structure, activity and stability, overview, deglycosylation by alpha-mannosidase does not affect the pH optimum or the active site and catalytic reaction mechanism including conformational changes of the enzyme, the secondary enzyme structure remains unaltered, deglycosylation increases the enzyme aggregation and reduces the thermal stability, the deglycosylated enzyme is more sensitive to proteolytic degradation by subtilisin than the native enzyme
glycoprotein
-
O-glycosylation, required for enzyme stability and activity, structure overview
proteolytic modification
-
the small enzyme form G2 consists of two molecular species identical to the residues 1-512 and 1-513 of the large enzyme form G1. The enzyme form G2 is generated by limited proteolysis of the larger enzyme form G1
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified isolated N-terminal catalytic domain of the G1 isoform, generated by subtilisin cleavage, sitting-drop vapour-diffusion method, 20 mg/ml protein at pH 8.5, 22°C room temperatur, the reservoir solution contains 50 mM Tris acetate, pH 8.5, 22.5% PEG 6000, 0.4 M sodium acetate and 10% glycerol, mixing of 0.001 ml protein and reservoir solution and equilibration against 1 ml reservoir solution, 2 weeks, X-ray diffraction structure determination and analysis at 1.9 A resolution, the active site of the enzyme is in complex with both Tris and glycerol molecules, structure modelling
purified recombinant enzyme, sitting drop vapour diffusion method, mixing of 150 nl of 18 mg/ml protein in 25 mM piperazine, pH 5.0, and 150 mM NaCl, with 150 nl of reservoir solution containing 0.1 M HEPES, pH 7.5, and 25% PEG 3350, 20°C, X-ray diffraction structure determination and analysis at 2.3 A resolution, molecular replacement using the catalytic domain of Aspergillus awamori glucoamylase as a search model (PDB entry 1glm)
purified starch-binding domain (SBD)-containing polypeptide (amino acids 509-616) of Aspergillus niger glucoamylase, hanging drop vapour diffusion method, using 2.0-2.3 M ammonium sulfate, 1.7% w/v PEG 400, 15% v/v glycerol, and 85 mM HEPES, pH 7.5, as the reservoir solution, the asymmetric unit of the starch-binding domain is composed of four protein molecules, 20°C, two weeks, X-ray diffraction structure determination and analysis at 2.0 A resolution, NMR structures of the SBD of Aspergillus niger glucoamylase (PDB entries 1kum, 1kul, 1aco, and 1acz) are used as search models for molecular replacement
cross linked glucoamylase crystals, glucoamylase is crystallized by the batch method using ammonium sulfate as precipitant, 65% saturation, along with 20% 2-propanol as cosolvent in 500 mM acetate buffer, pH 4.5, the solution is stirred at 4°C for 30 min and then kept for 16 h at the same temperature
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D20C/A27C/S30P/G137A
-
site-directed mutagenesis, the mutant, designated THS8, is highly thermotolerant with increased stability at 80°C compared to the wild-type enzyme
E180Q
H391Y
-
random mutagenesis, the mutant shows increased thermotolerance compared to the wild-type enzyme
R54L
-
active site mutant. For inhibitor acarbose, a rapid binding event is apparently intersected by a slower secondary binding event. Mutant shows a dramatically higher Kd value for acarbose
T290A
-
random mutagenesis, the mutant shows increased thermotolerance compared to the wild-type enzyme
T62A
-
random mutagenesis, the mutant shows increased thermotolerance compared to the wild-type enzyme
W120F
-
mutant of G1, 3% of wild-type kcat for maltose, 2% of kcat for maltotriose
W317F
-
mutant of G1, 90% of wild-type kcat for maltose, 97% of kcat for maltotriose
Y175F
-
mutation in subsite +3. Mutant displays only minor differences to wild-type in affinities to inhibitors acarbose and an acarbose conjugate
additional information
E180Q
-
mutant of G1, 48% of wild-type kcat for maltose, 88% of kcat for maltotriose
E180Q
-
active site mutant. For inhibitor acarbose, a rapid binding event is apparently intersected by a slower secondary binding event. Mutant shows a dramatically higher Kd value for acarbose
additional information
-
immobilization of the enzyme on polyaniline polymer results in improved catalytic performance with decreased temperature optimum, and increased thermal stability and catalytic efficiency with increased Vmax and reduced Km, overview
additional information
-
entrapment of amyloglucosidase into dipalmitoylphosphatidylcholine multilamellar vesicles and large unilamellar vesicles, vesicle formation, method optimization, enzyme activity is very stable during the first three batch runs, overview
additional information
-
random mutagenesis and mutant screening by plate thermostability assay for increased thermotolerance at 65-80°C, overview
additional information
-
engineered low-glycosylated variant of glucoamylase 1 with a short linker, low-glycosylated GA1 (dgGA). Low-glycosylated linker variant of GA1; GA1:L0 and dgGA:L0
additional information
-
disulfide-deficient mutant of the starch-binding domain of glucoamylase
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
-
enzyme in solid-state fermentation process of chestnut, pH 6.0, about 90% remaining activity after 24 h
50
60
60 - 80
-
the thermal stability at 70°C and other temperatures of the deglycosylated enzyme is reduced compared to the native enzyme
70
70 - 80
-
irreversible thermoinactivation kinetic analysis of recombinant wild-type and mutant enzymes, overview
80
90
-
crosslinked glucoamylase crystals, 48% loss of activity after 1 h in the presence of 4% starch, soluble glucoamylase, 55% loss of activity in the presence of 4% starch, complete loss of activity in the absence of starch
additional information
50
-
half-life of free enzyme is 33.8 h, half-life of immobilized enzyme is 190 h
50
-
pH 5.0, soluble enzyme half-life: 160 h, liposome-entrapped enzyme shows full activity after 180 h
70
-
250 min, immobilized enzyme loses 60% of its activity, half-life: 220 min. Free enzyme loses 75% of its activity, half-life 140 min
70
-
crosslinked glucoamylase crystals, 1.5% loss of activity after 1 h in the presence of 4% starch, soluble glucoamylase, 17% loss of activity in the presence of 4% starch, complete loss of activity in the absence of starch
80
-
crosslinked glucoamylase crystals, 23% loss of activity after 1 h in the presence of 4% starch, soluble glucoamylase, 38% loss of activity in the presence of 4% starch, complete loss of activity in the absence of starch
additional information
-
thermostability of immobilized glucoamylase increases considerably as a result of covalent immobilization onto poly(2-hydroxyethyl methacrylate)/ethylene glycol dimetharylate microspheres
additional information
-
the deglycosylated enzyme shows a higher aggregation rate compared to the native enzyme dependent on pH and presence of divalent cations, e.g. Mg2+ or Ca2+
additional information
-
the enzyme immobilized on polyaniline polymer shows an increased temperature stability compared to the free enzyme
additional information
-
during the thermal inactivation progress, combined with the loss of the helical structure and a majority of the tertiary structure, tryptophan residues are partially exposed and further lead to glucoamylases aggregating. The thermal stability of isoforms GAM-1 and GAM-2 is largely improved in the presence of sorbitol and trehalose by maintaining the native state of glucoamylases and preventing their thermal aggregation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
denaturation and renaturation does not seem to offer an economical approach to improve the usage time of immobilized enzyme
-
high operational stability of the enzyme immobilized onto poly(2-hydroxyethyl methacrylate)/ethylene glycol dimetharylate microspheres
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme from Aspergillus niger strain MBin118 by alpha-cyclodextrin affinity chromatography, elution with beta-cyclodextrin, followed by dialysis, to homogeneity
glucoamylase starch binding domain, affinity purification on granular corn starch
glucoamylases from a wild-type and a deoxy-D-glucose-resistant mutant Aspergillus niger to apparent homogeneity by ammonium sulfate fractionation, anion exchange chromatography, and hydrophobic interaction chromatography, the wild-tyype enzyme is purified 25.4fold, the mutant 30.6fold
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene glaA, DNA and amino acid sequence determination and analysis, the gene encoding glucoamylase G1 of Aspergillus niger strain DSM 823 shows high identity with the gene from Aspergillus tubingensis strains 643.92 and 127.49, phylogenetic analysis
gene GLAA, recombinant expression of the enzyme in Aspergillus niger strain MBin118
recombinant expression of the starch-binding domain (SBD)-containing polypeptide (amino acids 509-616) of glucoamylase in Escherichia coli
DNA and amino acid sequence determination and anaylsis of wild-type and mutant enzymes, expression of wild-type and mutant enzymes in Saccharomyces cerevisiae strain C468 using vector YEpPM18
-
expression of glucoamylase starch binding domain in Escherichia coli
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
industry
glucoamylase from Aspergillus niger is an industrially important biocatalyst that is utilized in the mass production of glucose from raw starch or soluble oligosaccharides
analysis
-
method for anchoring native acarbose to gold chip surfaces for surface plasmon resonance studies employing glucoamylase as analyte. The key method is the chemoselective and protecting group-free oxime functionalization of the pseudo-tetrasaccharide-based inhibitor acarbose. At pH 7.0 the association and dissociation rate constants for the acarbose-glucoamylase interaction are 10000 per M and s and 103 per s, respectively, and the conformational change to a tight enzyme-inhibitor complex affects the dissociation rate constant by a factor of 100 per s. The acarbose-presenting surface plason resonance surfaces can be used as a glucoamylase sensor that allows rapid, label-free affinity screening of small carbohydrate-based inhibitors in solution
biotechnology
-
immobilization of the enzyme on alginate beads for large-scale hydrolysis of starch in a fluidized bed of enzyme-alginate particles, method development, comparison to packed and batch mode, overview
industry
nutrition
synthesis
industry
-
immobilization of the enzyme on polyaniline polymer allows an improved catalytic performance and application of the enzyme in industrial processes
industry
-
glucoamylase is used industrially to catalyze the hydrolysis of alpha-1,4-glycosidic bonds to release beta-D-glucose from the nonreducing ends of starch or oligosaccharides
nutrition
-
optimization of solid-state enzymatic hydrolysis of chestnut in food production using the Aspergillus niger glucoamylase in concert with an alpha-amylase, overview
nutrition
-
the enzyme of the M115 mutant strain is useful for enhanced ethanol production by Saccharomyces cerevisiae, strain ATTC26602, using raw starch as substrate in solid state fermentation
nutrition
-
tannins isolated from extracts of pomegranate, cranberry, grape, and cocoa inhibit the activity of glucoamylase and alpha-amylase in vitro. In general, larger and more complex tannins, such as those in pomegranate and cranberry, more effectively inhibit the enzymes than less polymerized cocoa tannins
synthesis
-
the immobilized enzyme with high operational stability can be used for continuous production of glucose from soluble dextrin
synthesis
-
production of glucose, which is a feed stock for high fructose syrup
synthesis
-
the enzyme of the M115 mutant strain is useful for enhanced ethanol production by Saccharomyces cerevisiae, strain ATTC26602, using raw starch as substrate in solid state fermentation
synthesis
-
entrapment of amyloglucosidase into dipalmitoylphosphatidylcholine multilamellar vesicles and large unilamellar vesicles for biocatalysis inside liposomes and bioanalytical applications, overview
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Svensson, B.; Larsen, K.; Gunnarsson, A.
Characterization of a glucoamylase G2 from Aspergillus niger
Eur. J. Biochem.
154
497-502
1986
Aspergillus niger
Abe, J.I.; Nagano, H.; Hizukuri, S.
Kinetic and structural properties of the three forms of glucoamylase of Rhizopus delemar
J. Appl. Biochem.
7
235-247
1985
Aspergillus niger, Rhizopus arrhizus
-
Manjunath, P.; Shenoy, B.C.; Raghavendra Roa, M.R.
Fungal glucoamylases
J. Appl. Biochem.
5
235-260
1983
Aspergillus awamori, Aspergillus candidus, Aspergillus foetidus, Aspergillus niger, Aspergillus oryzae, Aspergillus phoenicis, Acremonium charticola, Amorphotheca resinae, Coniophora cerebella, Endomyces sp., Thermomyces lanuginosus, Pyricularia grisea, Mucor rouxianus, Rhizopus arrhizus
Fogarty, W.M.; Benson, C.P.
Purification and properties of a thermophilic amyloglucosidase from Aspergillus niger
Eur. J. Appl. Microbiol. Biotechnol.
18
271-278
1983
Aspergillus niger
-
Marshall, J.J.; Whelan, W.J.
Incomplete conversion of glycogen and starch by crystalline amyloglucosidase and its importance in the determination of amylaceous polymers
FEBS Lett.
9
85-88
1970
Aspergillus niger, Rhizopus niveus
Bahar, T.; Celebi, S.S.
Characterization of glucoamylase immobilized on magnetic poly(styrene) particles
Enzyme Microb. Technol.
23
301-304
1998
Aspergillus niger
-
Arica, M.Y.; Alaeddinoglu, N.G.; Hasirci, V.
Immobilization of glucoamylase onto activated pHEMA/EGDMA microspheres: properties and application to a packed-bed reactor
Enzyme Microb. Technol.
22
152-157
1998
Aspergillus niger
-
Gottschalk, N.; Jaenicke, R.
Authenticity and reconstitution of immobilized enzymes: characterization and denaturation/renaturation of glucoamylase II
Biotechnol. Appl. Biochem.
14
324-335
1991
Aspergillus niger
Goncalves, L.R.; Suzuki, G.S.; Giordano, R.C.; Giordano, R.L.
Kinetic and mass transfer parameters of maltotriose hydrolysis catalyzed by glucoamylase immobilized on macroporous silica and wrapped in pectin gel
Appl. Biochem. Biotechnol.
91-93
691-702
2001
Aspergillus niger
Christensen, U.
pH-dependence of the fast step of maltose hydrolysis catalysed by glucoamylase G1 from Aspergillus niger
Biochem. J.
349
623-628
2000
Aspergillus niger
Paldi, T.; Levy, I.; Shoseyov, O.
Glucoamylase starch-binding domain of Aspergillus niger B1: molecular cloning and functional characterization
Biochem. J.
372
905-910
2003
Aspergillus niger (Q8TG09), Aspergillus niger, Aspergillus niger B1 (Q8TG09), Aspergillus niger B1
Sauer, J.; Sigurskjold, B.W.; Christensen, U.; Frandsen, T.P.; Mirgorodskaya, E.; Harrison, M.; Roepstorff, P.; Svensson, B.
Glucoamylase: structure/function relationships, and protein engineering
Biochim. Biophys. Acta
1543
275-293
2000
Aspergillus awamori, Aspergillus niger, Saccharomycopsis fibuligera
Abraham, T.E.; Joseph, J.R.; Bindhu, L.B.V.; Jayakumar, K.K.
Crosslinked enzyme crystals of glucoamylase as a potent catalyst for biotransformations
Carbohydr. Res.
339
1099-1104
2004
Aspergillus niger
Roy, I.; Gupta, M.N.
Hydrolysis of starch by a mixture of glucoamylase and pullulanase entrapped individually in calcium alginate beads
Enzyme Microb. Technol.
34
26-32
2004
Aspergillus niger
-
Kainuma, K.
Applied glycoscience-past, present and future
Foods Food Ingredients J. Jpn.
178
4-10
1998
Aspergillus niger
-
Manger-Jacob, F.; Mueller, T.; Janssen, M.; Hoefer, M.; Hoelker, U.
Isolation and sequencing of a new glucoamylase gene from an Aspergillus niger aggregate strain (DSM 823) molecularly classified as Aspergillus tubingensis
Antonie van Leeuwenhoek
88
267-275
2005
Aspergillus tubingensis, Aspergillus niger (P69328), Aspergillus niger
Polakovic, M.; Bryjak, J.
Modelling of potato starch saccharification by an Aspergillus niger glucoamylase
Biochem. Eng. J.
18
57-63
2004
Aspergillus niger, Aspergillus niger A-7420
-
Jafari-Aghdam, J.; Khajeh, K.; Ranjbar, B.; Nemat-Gorgani, M.
Deglycosylation of glucoamylase from Aspergillus niger: effects on structure, activity and stability
Biochim. Biophys. Acta
1750
61-68
2005
Aspergillus niger
Norouzian, D.; Akbarzadeh, A.; Scharer, J.M.; Moo Young, M.
Fungal glucoamylases
Biotechnol. Adv.
24
80-85
2005
Aspergillus awamori, Aspergillus foetidus, Aspergillus niger, Aspergillus oryzae, Aspergillus phoenicis, Aspergillus terreus, Mucor javanicus, Neurospora crassa, Rhizopus arrhizus, Rhizopus niveus, Rhizopus microsporus var. oligosporus, Mycothermus thermophilus, Trichoderma reesei, Mucor rouxians, Arthrobotrys amerospora, Monascus kaoliang, Monascus kaoliang F-1, Aspergillus awamori X-100, Aspergillus terreus NA-170, Aspergillus niger NRRL 330, Aspergillus terreus NA-770
Lopez, C.; Torrado, A.; Guerra, N.P.; Pastrana, L.
Optimization of solid-state enzymatic hydrolysis of chestnut using mixtures of alpha-amylase and glucoamylase
J. Agric. Food Chem.
53
989-995
2005
Aspergillus niger
Rajoka, M.I.; Yasmin, A.; Latif, F.
Kinetics of enhanced ethanol productivity using raw starch hydrolyzing glucoamylase from Aspergillus niger mutant produced in solid state fermentation
Lett. Appl. Microbiol.
39
13-18
2004
Aspergillus niger
Silva, R.N.; Asquieri, E.R.; Fernandes, K.F.
Immobilization of Aspergillus niger glucoamylase onto a polyaniline polymer
Process Biochem.
40
1155-1159
2004
Aspergillus niger
-
Sutthirak, P.; Dharmsthiti, S.; Lertsiri, S.
Effect of glycation on stability and kinetic parameters of thermostable glucoamylase from Aspergillus niger
Process Biochem.
40
2821-2826
2005
Aspergillus niger
-
Rajoka, M.I.; Yasmeen, A.
Induction, and production studies of a novel glucoamylase of Aspergillus niger
World J. Microbiol. Biotechnol.
21
179-187
2005
Aspergillus niger, Aspergillus niger NIAB 280
-
Li, M.; Hanford, M.J.; Kim, J.W.; Peeples, T.L.
Amyloglucosidase enzymatic reactivity inside lipid vesicles
J. Biol. Eng.
1:4
no pp. given
2007
Aspergillus niger
Wang, Y.; Fuchs, E.; da Silva, R.; McDaniel, A.; Seibel, J.; Ford, C.
Improvement of Aspergillus niger glucoamylase thermostability by directed evolution
Starch Staerke
58
501-508
2006
Aspergillus niger
-
Jorgensen, A.D.; Nohr, J.; Kastrup, J.S.; Gajhede, M.; Sigurskjold, B.W.; Sauer, J.; Svergun, D.I.; Svensson, B.; Vestergaard, B.
Small angle X-ray studies reveal that Aspergillus niger glucoamylase has a defined extended conformation and can form dimers in solution
J. Biol. Chem.
283
14772-14780
2008
Aspergillus niger
Kumar, P.; Satyanarayana, T.
Microbial glucoamylases: characteristics and applications
Crit. Rev. Biotechnol.
29
225-255
2009
Aspergillus fumigatus, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Aspergillus phoenicis, Aspergillus sp., Aspergillus terreus, Aureobasidium pullulans, Saccharomyces cerevisiae, Moesziomyces antarcticus, Cephalosporium eichhorniae, Thermochaetoides thermophila, Thermoanaerobacter thermohydrosulfuricus, Thermoanaerobacterium thermosaccharolyticum, Curvularia lunata, Saccharomycopsis fibuligera, Endomycopsis fibuligera, Fusarium solani, Thermomyces lanuginosus, Humicola sp., Monascus sp. (in: Fungi), Mucor circinelloides, Mucor javanicus, Neurospora crassa, Paecilomyces variotii, Rhizopus arrhizus, Rhizopus sp., Saccharomyces cerevisiae 'var. diastaticus', Schwanniomyces castellii, Mycothermus thermophilus, Saccharolobus solfataricus, Thermoplasma acidophilum, Trichoderma reesei, Lactobacillus amylovorus, Thielaviopsis paradoxa, Thermomucor indicae-seudaticae, Picrophilus torridus, Arthrobotrys amerospora, Lentinula edodes L-54, Aspergillus awamori (Q12537)
Sugimoto, H.; Nakaura, M.; Nishimura, S.; Karita, S.; Miyake, H.; Tanaka, A.
Kinetically trapped metastable intermediate of a disulfide-deficient mutant of the starch-binding domain of glucoamylase
Protein Sci.
18
1715-1723
2009
Aspergillus niger
Lee, J.; Paetzel, M.
Structure of the catalytic domain of glucoamylase from Aspergillus niger
Acta Crystallogr. Sect. F
67
188-192
2011
Aspergillus niger (P69328), Aspergillus niger
Riaz, M.; Rashid, M.; Sawyer, L.; Akhtar, S.; Javed, M.; Nadeem, H.; Wear, M.
Physiochemical properties and kinetics of glucoamylase produced from deoxy-D-glucose resistant mutant of Aspergillus niger for soluble starch hydrolysis
Food Chem.
130
24-30
2012
Aspergillus niger
Sauer, J.; Abou Hachem, M.; Svensson, B.; Jensen, K.J.; Thygesen, M.B.
Kinetic analysis of inhibition of glucoamylase and active site mutants via chemoselective oxime immobilization of acarbose on SPR chip surfaces
Carbohydr. Res.
375
21-28
2013
Aspergillus niger
Bagheri, A.; Khodarahmi, R.; Mostafaie, A.
Purification and biochemical characterisation of glucoamylase from a newly isolated Aspergillus niger: relation to starch processing
Food Chem.
161
270-278
2014
Aspergillus niger
Barrett, A.; Ndou, T.; Hughey, C.A.; Straut, C.; Howell, A.; Dai, Z.; Kaletunc, G.
Inhibition of alpha-amylase and glucoamylase by tannins extracted from cocoa, pomegranates, cranberries, and grapes
J. Agric. Food Chem.
61
1477-1486
2013
Aspergillus niger
Liu, Y.; Meng, Z.; Shi, R.; Zhan, L.; Hu, W.; Xiang, H.; Xie, Q.
Effects of temperature and additives on the thermal stability of glucoamylase from Aspergillus niger
J. Microbiol. Biotechnol.
24
33-43
2014
Aspergillus niger
Roth, C.; Moroz, O.V.; Ariza, A.; Skov, L.K.; Ayabe, K.; Davies, G.J.; Wilson, K.S.
Structural insight into industrially relevant glucoamylases flexible positions of starch-binding domains
Acta Crystallogr. Sect. D
74
463-470
2018
Aspergillus niger (P69328), Amorphotheca resinae (Q03045), Penicillium oxalicum (S7ZIW0), Penicillium oxalicum 114-2 (S7ZIW0), Penicillium oxalicum CGMCC 5302 (S7ZIW0)
Suyama, Y.; Muraki, N.; Kusunoki, M.; Miyake, H.
Crystal structure of the starch-binding domain of glucoamylase from Aspergillus niger
Acta Crystallogr. Sect. F
73
550-554
2017
Aspergillus niger (P69328), Aspergillus niger
EXTERNAL LINKS (specific for EC-Number 3.2.1.3)
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
