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Information on EC 3.2.1.1 - alpha-amylase and Organism(s) Pyrococcus furiosus and UniProt Accession O08452

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IUBMB Comments
Acts on starch, glycogen and related polysaccharides and oligosaccharides in a random manner; reducing groups are liberated in the alpha-configuration. The term "alpha" relates to the initial anomeric configuration of the free sugar group released and not to the configuration of the linkage hydrolysed.
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Pyrococcus furiosus
UNIPROT: O08452
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The taxonomic range for the selected organisms is: Pyrococcus furiosus
The enzyme appears in selected viruses and cellular organisms
Synonyms
alpha-amylase, diastase, alpha amylase, pancreatic alpha-amylase, crustacean cardioactive peptide, maltogenic amylase, taka-amylase a, human salivary alpha-amylase, bacillus licheniformis alpha-amylase, alpha-amylase 2, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1,4-alpha-D-glucan glucanohydrolase
-
-
-
-
alpha-1,4-glucan-4-glucanohydrolase
-
-
Alpha-amylase carcinoid
-
-
-
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Amy c6
-
-
-
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AMY1
-
-
-
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Amylase THC 250
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-
-
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amylase, alpha-
-
-
-
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Amylopsin
-
-
-
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Bactosol TK
-
-
-
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Buclamase
-
-
-
-
Clarase
-
-
-
-
Clone 103
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-
-
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Clone 168
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-
-
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Clone PHV19
-
-
-
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Clones GRAMY56 and 963
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-
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diastase
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-
-
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endoamylase
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-
-
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Fortizyme
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-
-
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G 995
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-
-
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glycogenase
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-
-
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High pI alpha-amylase
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-
-
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Isozyme 1B
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-
-
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Kleistase L 1
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-
-
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Low pI alpha-amylase
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-
-
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Maxamyl
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-
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Maxilase
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-
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Meiotic expression upregulated protein 30
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-
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Pancreatic alpha-amylase
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-
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Pivozin
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-
-
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Ptyalin
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-
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Spitase CP 1
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-
-
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TAA
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-
-
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Taka-amylase A
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-
-
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Takatherm
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-
-
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Thermamyl
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-
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Thermolase
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-
-
-
additional information
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cf. EC 3.2.1.54, cyclomaltodextrinase, the enzyme belongs to glycoside hydrolase family 13
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Endohydrolysis of (1->4)-alpha-D-glucosidic linkages in polysaccharides containing three or more (1->4)-alpha-linked D-glucose units
show the reaction diagram
PFTA is a bifunctional enzyme showing alpha-amylase as well as cyclodextrin-hydrolyzing activity
-
SYSTEMATIC NAME
IUBMB Comments
4-alpha-D-glucan glucanohydrolase
Acts on starch, glycogen and related polysaccharides and oligosaccharides in a random manner; reducing groups are liberated in the alpha-configuration. The term "alpha" relates to the initial anomeric configuration of the free sugar group released and not to the configuration of the linkage hydrolysed.
CAS REGISTRY NUMBER
COMMENTARY hide
9000-90-2
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SUBSTRATE
PRODUCT                       
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
liquefying enzyme. The main products of polysaccharide hydrolysis are G2 to G7. A small amount of G1 is formed after long hydrolysis periods. The enzyme A hydrolyzes long-chain oligosaccharides faster than shorter chain oligosaccharides
-
-
?
amylose + H2O
?
show the reaction diagram
liquefying enzyme. The main products of polysaccharide hydrolysis are G2 to G7. A small amount of G1 is formed after long hydrolysis periods. The enzyme A hydrolyzes long-chain oligosaccharides faster than shorter chain oligosaccharides
-
-
?
glycogen + H2O
?
show the reaction diagram
liquefying enzyme. The main products of polysaccharide hydrolysis are G2 to G7. A small amount of G1 is formed after long hydrolysis periods. The enzyme A hydrolyzes long-chain oligosaccharides faster than shorter chain oligosaccharides
-
-
?
maltodextran + H2O
?
show the reaction diagram
-
-
-
?
maltodextrin + H2O
?
show the reaction diagram
white dextrin
-
-
?
soluble starch + H2O
?
show the reaction diagram
4-nitrophenyl-alpha-D-maltopentaoside + H2O
4-nitrophenol + alpha-D-maltopentaoside
show the reaction diagram
-
-
-
-
?
acarbose + H2O
?
show the reaction diagram
-
the substrate is a potent inhibitor of alpha-amylases, cyclomaltodextrinase activity
-
-
?
alpha-cyclodextrin + H2O
?
show the reaction diagram
-
cyclomaltodextrinase activity
-
-
?
beta-cyclodextrin + H2O
panose + maltoheptaose
show the reaction diagram
-
cyclomaltodextrinase activity
-
-
?
gamma-cyclodextrin + H2O
?
show the reaction diagram
-
cyclomaltodextrinase activity
-
-
?
glycogen + H2O
?
show the reaction diagram
-
-
-
-
?
maltooligosaccharides + H2O
maltotriose + maltotetraose
show the reaction diagram
-
alpha-amylase activity
main products
-
?
maltotriose
maltose + glucose + maltotetraose + maltopentaose + maltohexaose
show the reaction diagram
-
the final optimum also mirrors the reverse reaction
-
r
maltotriose + H2O
?
show the reaction diagram
-
very low activity
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-
?
maltotriose + H2O
maltose + D-glucose
show the reaction diagram
-
-
-
-
?
pullulan + H2O
panose + maltoheptaose
show the reaction diagram
-
cyclomaltodextrinase activity
-
-
?
starch + H2O
?
show the reaction diagram
-
-
-
-
?
starch + H2O
malto-oligosaccharides
show the reaction diagram
-
-
-
-
?
starch + H2O
maltotriose + maltotetraose
show the reaction diagram
-
alpha-amylase activity, preferred substrate
main products
-
?
amylopectin + H2O
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
glycogen + H2O
?
show the reaction diagram
-
-
-
-
?
starch + H2O
malto-oligosaccharides
show the reaction diagram
-
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Cu2+
5 mM, complete inhibition
Ni2+
slight inhibition
Zn2+
5 mM, complete inhibition
alpha-cyclodextrin
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1-10 mM, 25-35% inhibition
beta-cyclodextrin
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1-10 mM, 10-25% inhibition
EDTA
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1-10 mM, about 60-65% inhibition
EGTA
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1-10 mM, about 50% inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
11.1
acarbose
-
pH 4.5, 90°C, recombinant enzyme
2.61
alpha-cyclodextrin
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pH 4.5, 90°C, recombinant enzyme
2.16
beta-cyclodextrin
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pH 4.5, 90°C, recombinant enzyme
5.12
gamma-cyclodextrin
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pH 4.5, 90°C, recombinant enzyme
62.9
maltotriose
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pH 4.5, 90°C, recombinant enzyme
0.52
starch
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pH 4.5, 90°C, recombinant enzyme
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
228
acarbose
-
pH 4.5, 90°C, recombinant enzyme
241
alpha-cyclodextrin
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pH 4.5, 90°C, recombinant enzyme
196
beta-cyclodextrin
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pH 4.5, 90°C, recombinant enzyme
173
gamma-cyclodextrin
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pH 4.5, 90°C, recombinant enzyme
268
maltotriose
-
pH 4.5, 90°C, recombinant enzyme
67
starch
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pH 4.5, 90°C, recombinant enzyme
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3900
pH 6.0, at 98°C
176.4
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purified recombinant enzyme, substrate beta-cyclodextrin
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5
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assay at
6.5 - 7.5
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5 - 7.5
pH 3.5: about 70% of maximal activity, pH 7.5: about 40% of maximal activity
4.7 - 7
80% activity or more between pH 4.7 and pH 7.0
6 - 8
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pH 6.0: about 50% of maximal activity, pH 8.0: about 60% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
no activity at room temperature
90
-
recombinant enzyme
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
70 - 130
70°C: about 55% of maximal activity, 130°C: about 65% of maximal activity
70 - 115
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70°C: about 30% of maximal activity, 115°C: about 75% of maximal activity
90 - 109
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activity at 90°C is about 60% compared to the activity at 108°C
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?
O08452_9EURY
460
1
52910
TrEMBL
-
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
100000
gel filtration
52875
x * 52875, extracellular enzyme, calculation from nucleotide sequence
66000
2 * 66000, SDS-PAGE
129000
-
non-denaturing PAGE
130500
-
equilibrium ultracentrifugation
157000
-
gel filtration
66000
-
2 * 66000, intracellular enzyme, electrophoresis in presence of 8 M urea
76084
-
x * 76084, amino acid sequence calculation
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 52875, extracellular enzyme, calculation from nucleotide sequence
homodimer
2 * 66000, SDS-PAGE
?
-
x * 76084, amino acid sequence calculation
dimer
-
2 * 66000, intracellular enzyme, electrophoresis in presence of 8 M urea
additional information
-
primary structure scheme
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
100
6 h, 20% loss of activity
115
3 h, 65% loss of activity
120
1 h, 40% loss of activity
98
half-life: 13 min
100
-
purified recombinant enzyme, sodium acetate buffer, pH 5.0, loss of 65% activity within 60 min
105
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2 h, about 80% loss of activity
106
-
denaturation of the enzyme is irreversible above a Tm of approximately 106°C and can be described by a one-step irreversible model
117
-
t1/2: 10.1 min
121
-
t1/2: 6.1 min, activation energy at 121°C is 316kJ/mol. Under an isothermal holding temperature of 121°C, the structure of the enzyme changes during denaturation from an alpha-helical structure, through a beta-sheet structure to an aggregated protein
125
-
t1/2: 2.7 min
85
-
purified recombinant enzyme, sodium acetate buffer, pH 5.0, completely stable for 120 min
90
-
purified recombinant enzyme, sodium acetate buffer, pH 5.0, loss of 15-20% activity within 120 min
95
-
purified recombinant enzyme, sodium acetate buffer, pH 5.0, loss of 25% activity within 120 min
98
-
8 h, about 70% loss of activity
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
free Ca2+, slight stabilization
-
guanidine HCl, 1 M, 27% loss of activity, significant decrease in activity at 2 M
-
moderate protease susceptibility
-
urea, 1 M, 13% loss of activity, significant decrease in activity at 2 M
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme expressed as inclusion bodies in Escherichia coli. The solubilization of the inclusion bodies is achieved by 90°C treatment for 3 min in Britton-Robinson buffer (0.04 M H3BO3, 0.04 M H3PO4, 0.04 M CH3COOH) at pH 10.5. The solubilized PFA is then diluted and subsequently purified by Phenyl Sepharose chromatography
recombinant PFTA 32.6fold from Escherichia coli
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression as inclusion bodies in Escherichia coli
expression in Escherichia coli
expression in Escherichia coli and Bacillus subtilis
expression in Chlamydomonas reinhardtii chloroplast
-
PTFA, DNA and amino acid sequence determination and analysis, expression in Escherichia coli strain BL21(DE3) and MC1061
-
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
heat denaturation is irreversible
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
brewing
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important industrial enzyme in brewing and alcohol production
food industry
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important industrial enzyme in brewing and alcohol production
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Laderman, K.A.; Davis, B.R.; Krutzsch, H.C.; Lewis, M.S.; Griko, Y.V.; Privalov, P.L.; Anfinsen, C.B.
The purification and characterization of an extremely thermostable alpha-amylase from the hyperthermophilic archaebacterium Pyrococcus furiosus
J. Biol. Chem.
268
24394-24401
1993
Pyrococcus furiosus
Manually annotated by BRENDA team
Jorgensen, S.; Vorgias, C.E.; Antranikian G.
Cloning, sequencing, characterization, and expression of an extracellular alpha-amylase from the hyperthermophilic archaeon Pyrococcus furiosus in Escherichia coli and Bacillus subtilis
J. Biol. Chem.
272
16335-16342
1997
Pyrococcus furiosus (O08452), Pyrococcus furiosus
Manually annotated by BRENDA team
Yang, S.J.; Lee, H.S.; Park, C.S.; Kim, Y.R.; Moon, T.W.; Park, K.H.
Enzymatic analysis of an amylolytic enzyme from the hyperthermophilic archaeon Pyrococcus furiosus reveals its novel catalytic properties as both an alpha-amylase and a cyclodextrin-hydrolyzing enzyme
Appl. Environ. Microbiol.
70
5988-5995
2004
Pyrococcus furiosus
Manually annotated by BRENDA team
Wang, L.; Zhou, Q.; Chen, H.; Chu, Z.; Lu, J.; Zhang, Y.; Yang, S.
Efficient solubilization, purification of recombinant extracellular alpha-amylase from Pyrococcus furiosus expressed as inclusion bodies in Escherichia coli
J. Ind. Microbiol. Biotechnol.
34
187-192
2007
Pyrococcus furiosus (O08452), Pyrococcus furiosus
Manually annotated by BRENDA team
Prakash, O.; Jaiswal, N.
alpha-Amylase: an ideal representative of thermostable enzymes
Appl. Biochem. Biotechnol.
160
2401-2414
2010
Alicyclobacillus acidocaldarius, Geobacillus stearothermophilus, Bacillus amyloliquefaciens, Anoxybacillus flavithermus, Bacillus subtilis, Lederbergia lentus, Bacillus licheniformis, Chloroflexus aurantiacus, Thermothelomyces heterothallicus, Desulfurococcus mucosus, Dictyoglomus thermophilum, Thermomyces lanuginosus, Lactiplantibacillus plantarum, Lipomyces kononenkoae, Pyrococcus furiosus, Pyrococcus woesei, Pyrodictium abyssi, Rhizopus sp., Rhodothermus marinus, Mycothermus thermophilus, Staphylothermus marinus, Thermoactinomyces vulgaris, Thermococcus fumicolans, Thermococcus hydrothermalis, Thermococcus litoralis, Thermococcus profundus, Thermotoga maritima, Thermus filiformis, Lactobacillus amylovorus, Halothermothrix orenii, Thermococcus aggregans, Thermococcus celer, Thermococcus guaymasensis
Manually annotated by BRENDA team
Brown, S.H.; Costantino, H.R.; Kelly, R.M.
Characterization of amylolytic enzyme activities associated with the hyperthermophilic archaebacterium Pyrococcus furiosus
Appl. Environ. Microbiol.
56
1985-1991
1990
Pyrococcus furiosus
Manually annotated by BRENDA team
Dong, G.; Vieille, C.; Savchenko, A.; Zeikus, J.
Cloning, sequencing, and expression of the gene encoding extracellular alpha-amylase from Pyrococcus furiosus and biochemical characterization of the recombinant enzyme
Appl. Environ. Microbiol.
63
3569-3576
1997
Pyrococcus furiosus (O08452), Pyrococcus furiosus
Manually annotated by BRENDA team
Brown, I.; Dafforn, T.R.; Fryer, P.J.; Cox, P.W.
Kinetic study of the thermal denaturation of a hyperthermostable extracellular alpha-amylase from Pyrococcus furiosus
Biochim. Biophys. Acta
1834
2600-2605
2013
Pyrococcus furiosus
Manually annotated by BRENDA team
Yang, Z.Q.; Li, Y.N. Zhang, Z.F.; Wang, Y.; Shen, G.F.
Expression of the gene coding for a thermostable alpha-amylase from Pyrococcus furious in Chiamydomonas reinhardtii chloroplast
Sheng Wu Gong Cheng Xue Bao
22
545-549
2006
Pyrococcus furiosus
Manually annotated by BRENDA team