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Literature summary extracted from

  • Park, K.M.; Jun, S.Y.; Choi, K.H.; Park, K.H.; Park, C.S.; Cha, J.
    Characterization of an exo-acting intracellular alpha-amylase from the hyperthermophilic bacterium Thermotoga neapolitana (2010), Appl. Microbiol. Biotechnol., 86, 555-566.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.1 expressed in Escherichia coli BL21(DE3) cells Thermotoga neapolitana

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.1 Co2+ complete inhibition at 1 mM Thermotoga neapolitana
3.2.1.1 Cu2+ 52.2% residual activity at 1 mM Thermotoga neapolitana
3.2.1.1 Hg2+ complete inhibition at 1 mM Thermotoga neapolitana
3.2.1.1 Mg2+ 69.4% residual activity at 1 mM Thermotoga neapolitana
3.2.1.1 Mn2+ complete inhibition at 1 mM Thermotoga neapolitana
3.2.1.1 additional information EDTA, dithiothreitol, N-bromosuccinimide, 2-mercaptoethanol (1 mM), and SDS (2% (w/v)) have no effect on activity Thermotoga neapolitana
3.2.1.1 Urea 25.8% residual activity at 1 mM Thermotoga neapolitana
3.2.1.1 Zn2+ 70.4% residual activity at 1 mM Thermotoga neapolitana

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.1 0.07
-
soluble starch in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana
3.2.1.1 1.26
-
maltotriose in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana
3.2.1.1 1.69
-
maltopentaose in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana
3.2.1.1 2.46
-
4-nitrophenyl alpha-D-maltohexaoside in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana
3.2.1.1 3.37
-
maltoheptaose in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.2.1.1 intracellular
-
Thermotoga neapolitana 5622
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.1 additional information Ni2+ and Ca2+ do not stimulate the activity of AmyB Thermotoga neapolitana

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.1 48000
-
SDS-PAGE and gel filtration Thermotoga neapolitana
3.2.1.1 50120
-
calculated from amino acid sequence Thermotoga neapolitana

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.1 4-nitrophenyl alpha-D-maltohexaoside + H2O Thermotoga neapolitana
-
?
-
?
3.2.1.1 amylase + H2O Thermotoga neapolitana
-
?
-
?
3.2.1.1 amylopectin + H2O Thermotoga neapolitana
-
?
-
?
3.2.1.1 glycogen + H2O Thermotoga neapolitana
-
?
-
?
3.2.1.1 glycogen + H2O Thermotoga neapolitana 41025
-
?
-
?
3.2.1.1 maltoheptaose + H2O Thermotoga neapolitana
-
maltose + D-glucose + ?
-
?
3.2.1.1 maltohexaose + H2O Thermotoga neapolitana
-
3 maltose
-
?
3.2.1.1 maltopentaose + H2O Thermotoga neapolitana
-
maltose + D-glucose + ?
-
?
3.2.1.1 maltotetraose + H2O Thermotoga neapolitana
-
2 maltose
-
?
3.2.1.1 maltotetraose + H2O Thermotoga neapolitana 41025
-
2 maltose
-
?
3.2.1.1 maltotriose + H2O Thermotoga neapolitana
-
maltose + D-glucose
-
?
3.2.1.1 additional information Thermotoga neapolitana AmyB enzymatic activity is negligible when acarbose, a maltotetraose is present, indicating that AmyB cleaves maltose units from the nonreducing end of maltooligosaccharides ?
-
?
3.2.1.1 additional information Thermotoga neapolitana inability to hydrolyze pullulan and beta-cyclodextrin with less than 10% of the specific activity of starch ?
-
?
3.2.1.1 additional information Thermotoga neapolitana 41025 AmyB enzymatic activity is negligible when acarbose, a maltotetraose is present, indicating that AmyB cleaves maltose units from the nonreducing end of maltooligosaccharides ?
-
?
3.2.1.1 additional information Thermotoga neapolitana 41025 inability to hydrolyze pullulan and beta-cyclodextrin with less than 10% of the specific activity of starch ?
-
?
3.2.1.1 soluble starch + H2O Thermotoga neapolitana
-
?
-
?
3.2.1.1 soluble starch + H2O Thermotoga neapolitana 41025
-
?
-
?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.1 Thermotoga neapolitana B5ARZ9
-
-
3.2.1.1 Thermotoga neapolitana 41025 B5ARZ9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.1 HiTrap SP column chromatography Thermotoga neapolitana

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.1 0.0063
-
using amylopectin as substrate, in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana
3.2.1.1 0.0109
-
using amylose as substrate, in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana
3.2.1.1 0.0188
-
using soluble starch as substrate, in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.1 4-nitrophenyl alpha-D-maltohexaoside + H2O
-
Thermotoga neapolitana ?
-
?
3.2.1.1 amylase + H2O
-
Thermotoga neapolitana ?
-
?
3.2.1.1 amylopectin + H2O
-
Thermotoga neapolitana ?
-
?
3.2.1.1 glycogen + H2O
-
Thermotoga neapolitana ?
-
?
3.2.1.1 glycogen + H2O
-
Thermotoga neapolitana 41025 ?
-
?
3.2.1.1 maltoheptaose + H2O
-
Thermotoga neapolitana maltose + D-glucose + ?
-
?
3.2.1.1 maltohexaose + H2O
-
Thermotoga neapolitana 3 maltose
-
?
3.2.1.1 maltopentaose + H2O
-
Thermotoga neapolitana maltose + D-glucose + ?
-
?
3.2.1.1 maltotetraose + H2O
-
Thermotoga neapolitana 2 maltose
-
?
3.2.1.1 maltotetraose + H2O
-
Thermotoga neapolitana 41025 2 maltose
-
?
3.2.1.1 maltotriose + H2O
-
Thermotoga neapolitana maltose + D-glucose
-
?
3.2.1.1 additional information AmyB enzymatic activity is negligible when acarbose, a maltotetraose is present, indicating that AmyB cleaves maltose units from the nonreducing end of maltooligosaccharides Thermotoga neapolitana ?
-
?
3.2.1.1 additional information inability to hydrolyze pullulan and beta-cyclodextrin with less than 10% of the specific activity of starch Thermotoga neapolitana ?
-
?
3.2.1.1 additional information AmyB enzymatic activity is negligible when acarbose, a maltotetraose is present, indicating that AmyB cleaves maltose units from the nonreducing end of maltooligosaccharides Thermotoga neapolitana 41025 ?
-
?
3.2.1.1 additional information inability to hydrolyze pullulan and beta-cyclodextrin with less than 10% of the specific activity of starch Thermotoga neapolitana 41025 ?
-
?
3.2.1.1 soluble starch + H2O
-
Thermotoga neapolitana ?
-
?
3.2.1.1 soluble starch + H2O
-
Thermotoga neapolitana 41025 ?
-
?

Subunits

EC Number Subunits Comment Organism
3.2.1.1 monomer 1 * 48000, gel filtration Thermotoga neapolitana

Synonyms

EC Number Synonyms Comment Organism
3.2.1.1 AmyB exo-acting intracellular alpha-amylase that possesses distinct characteristics from typical alpha-amylases and cyclodextrin-hydrolyzing enzymes such as maltogenic amylase, cyclodextrinase, and neopullulanase Thermotoga neapolitana

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.1 75
-
-
Thermotoga neapolitana

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.1 90 100 the enzyme retains its full activity after 7 h of incubation at 90°C, the half-life of AmyB is determined to be 50.2 and 28 h at 95°C and 100°C, respectively, thermostability is slightly enhanced by Ca2+ Thermotoga neapolitana

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.1 0.02
-
soluble starch in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana
3.2.1.1 25
-
maltoheptaose in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana
3.2.1.1 34
-
4-nitrophenyl alpha-D-maltohexaoside in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana
3.2.1.1 283
-
maltopentaose in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana
3.2.1.1 809
-
maltotriose in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.1 6.5
-
-
Thermotoga neapolitana

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.1 5 8.5
-
Thermotoga neapolitana

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.1 0.286
-
soluble starch in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana
3.2.1.1 7.5
-
maltoheptaose in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana
3.2.1.1 13.9
-
4-nitrophenyl alpha-D-maltohexaoside in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana
3.2.1.1 167.5
-
maltopentaose in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana
3.2.1.1 642.1
-
maltotriose in 50 mM sodium phosphate buffer (pH 6.5) at 75°C Thermotoga neapolitana