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Literature summary for 3.2.1.54 extracted from

  • 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 (2004), Appl. Environ. Microbiol., 70, 5988-5995.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.2.1.54

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Pyrococcus furiosus
PTFA, DNA and amino acid sequence determination and analysis, expression in Escherichia coli strain BL21(DE3) and MC1061 Pyrococcus furiosus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetics Pyrococcus furiosus
0.52
-
 starch pH 4.5, 90°C, recombinant enzyme Pyrococcus furiosus
2.16
-
 beta-cyclodextrin pH 4.5, 90°C Pyrococcus furiosus
2.16
-
 beta-cyclodextrin pH 4.5, 90°C, recombinant enzyme Pyrococcus furiosus
2.61
-
 alpha-cyclodextrin pH 4.5, 90°C Pyrococcus furiosus
2.61
-
 alpha-cyclodextrin pH 4.5, 90°C, recombinant enzyme Pyrococcus furiosus
5.12
-
 gamma-cyclodextrin pH 4.5, 90°C Pyrococcus furiosus
5.12
-
 gamma-cyclodextrin pH 4.5, 90°C, recombinant enzyme Pyrococcus furiosus
11.1
-
 acarbose pH 4.5, 90°C Pyrococcus furiosus
11.1
-
 acarbose pH 4.5, 90°C, recombinant enzyme Pyrococcus furiosus
62.9
-
 maltotriose pH 4.5, 90°C Pyrococcus furiosus
62.9
-
 maltotriose pH 4.5, 90°C, recombinant enzyme Pyrococcus furiosus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
76084
-
 x * 76084, amino acid sequence calculation Pyrococcus furiosus
76084
-
 x * 76084, calculated from sequence Pyrococcus furiosus

Organism

Organism UniProt Comment Textmining
Pyrococcus furiosus
-
-
-
Pyrococcus furiosus
-
 bifunctional enzyme
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pyrococcus furiosus
recombinant PFTA 32.6fold from Escherichia coli Pyrococcus furiosus

Reaction

Reaction Comment Organism Reaction ID
cyclomaltodextrin + H2O = linear maltodextrin PFTA is a bifunctional enzyme showing alpha-amylase as well as cyclodextrin-hydrolyzing activity Pyrococcus furiosus
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
 Pyrococcus furiosus
176.4
-
-
 Pyrococcus furiosus
176.4
-
 purified recombinant enzyme, substrate beta-cyclodextrin Pyrococcus furiosus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl-alpha-D-maltopentaoside + H2O
-
 Pyrococcus furiosus 4-nitrophenol + alpha-D-maltopentaoside
-
 ?
acarbose + H2O the substrate is a potent inhibitor of alpha-amylases, cyclomaltodextrinase activity Pyrococcus furiosus ?
-
 ?
acarbose + H2O
-
 Pyrococcus furiosus acarviosine-glucose + alpha-D-glucose
-
 ?
alpha-cyclodextrin + H2O
-
 Pyrococcus furiosus ?
-
 ?
alpha-cyclodextrin + H2O cyclomaltodextrinase activity Pyrococcus furiosus ?
-
 ?
beta-cyclodextrin + H2O
-
 Pyrococcus furiosus maltooligosaccharides G1 to G7, maltoheptaose as major product ?
beta-cyclodextrin + H2O cyclomaltodextrinase activity Pyrococcus furiosus panose + maltoheptaose
-
 ?
gamma-cyclodextrin + H2O
-
 Pyrococcus furiosus ?
-
 ?
gamma-cyclodextrin + H2O cyclomaltodextrinase activity Pyrococcus furiosus ?
-
 ?
maltooligosaccharides + H2O alpha-amylase activity Pyrococcus furiosus maltotriose + maltotetraose main products ?
maltotriose + H2O
-
 Pyrococcus furiosus ?
-
 ?
maltotriose + H2O very low activity Pyrococcus furiosus ?
-
 ?
additional information substrate specificity, hydrolytic pattern, PFTA is a bifunctional enzyme showing alpha-amylase as well as cyclodextrin-hydrolyzing activity, but no transglycosylation activity, overview Pyrococcus furiosus ?
-
 ?
pullulan + H2O
-
 Pyrococcus furiosus panose + beta-cyclodextrin
-
 ?
pullulan + H2O cyclomaltodextrinase activity Pyrococcus furiosus panose + maltoheptaose
-
 ?
soluble starch + H2O
-
 Pyrococcus furiosus ?
-
 ?
starch + H2O alpha-amylase activity, preferred substrate Pyrococcus furiosus maltotriose + maltotetraose main products ?

Subunits

Subunits Comment Organism
? x * 76084, amino acid sequence calculation Pyrococcus furiosus
? x * 76084, calculated from sequence Pyrococcus furiosus
More primary structure scheme Pyrococcus furiosus

Synonyms

Synonyms Comment Organism
More cf. EC 3.2.1.1, alpha-amylase, the enzyme belongs to glycoside hydrolase family 13 Pyrococcus furiosus
PFTA
-
 Pyrococcus furiosus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
90
-
-
 Pyrococcus furiosus
90
-
 recombinant enzyme Pyrococcus furiosus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
70 100 70°C: about 60% of maximal activity, 95°C: about 70% of maximal activity, 100°C: about 20% of maximal activity Pyrococcus furiosus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
 highly thermostable recombinant enzyme Pyrococcus furiosus
85
-
 120 min, stable Pyrococcus furiosus
85
-
 purified recombinant enzyme, sodium acetate buffer, pH 5.0, completely stable for 120 min Pyrococcus furiosus
90
-
 half-life: 398 min Pyrococcus furiosus
90
-
 purified recombinant enzyme, sodium acetate buffer, pH 5.0, loss of 15-20% activity within 120 min Pyrococcus furiosus
95
-
 half-life: 114 min Pyrococcus furiosus
95
-
 purified recombinant enzyme, sodium acetate buffer, pH 5.0, loss of 25% activity within 120 min Pyrococcus furiosus
100
-
 1 h, more than 27% of initial activity remains Pyrococcus furiosus
100
-
 purified recombinant enzyme, sodium acetate buffer, pH 5.0, loss of 65% activity within 60 min Pyrococcus furiosus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
67
-
 starch pH 4.5, 90°C, recombinant enzyme Pyrococcus furiosus
173
-
 gamma-cyclodextrin pH 4.5, 90°C Pyrococcus furiosus
173
-
 gamma-cyclodextrin pH 4.5, 90°C, recombinant enzyme Pyrococcus furiosus
196
-
 beta-cyclodextrin pH 4.5, 90°C Pyrococcus furiosus
196
-
 beta-cyclodextrin pH 4.5, 90°C, recombinant enzyme Pyrococcus furiosus
228
-
 acarbose pH 4.5, 90°C Pyrococcus furiosus
228
-
 acarbose pH 4.5, 90°C, recombinant enzyme Pyrococcus furiosus
241
-
 alpha-cyclodextrin pH 4.5, 90°C Pyrococcus furiosus
241
-
 alpha-cyclodextrin pH 4.5, 90°C, recombinant enzyme Pyrococcus furiosus
268
-
 maltotriose pH 4.5, 90°C Pyrococcus furiosus
268
-
 maltotriose pH 4.5, 90°C, recombinant enzyme Pyrococcus furiosus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
4.5
-
-
 Pyrococcus furiosus
4.5
-
 assay at Pyrococcus furiosus

pH Range

pH Minimum pH Maximum Comment Organism
4 6 about 50% of maximal activity at pH 4.0 and 6.0 Pyrococcus furiosus

pH Stability

pH Stability pH Stability Maximum Comment Organism
2.5
-
 37°C, 24 h, complete inactivation Pyrococcus furiosus
3
-
 37°C, 24 h, 20% loss of activity Pyrococcus furiosus
4 11 37°C, 24 h, stable Pyrococcus furiosus