Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Tang, K.; Kobayashi, R.S.; Champreda, V.; Eurwilaichitr, L.; Tanapongpipat, S.
    Isolation and characterization of a novel thermostable neopullulanase-like enzyme from a hot spring in Thailand (2008), Biosci. Biotechnol. Biochem., 72, 1448-1456.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.135 Escherichia coli DH5alphaF' grown in Luria-Bertani broth, 37°C with plasmid pTZ57R/T, expression in yeast Pichia pastoris with recombinant plasmid pPICZ-BK193 in Yeast Extract Peptone Dextrose uncultured microorganism

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.135 1.22
-
 pullulan optimal pH 7.0, optimal temperature 75°C uncultured microorganism

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.135 Ca2+ essential for enzyme activity, also tested were Mg2+, Sr2+, Zn2+, Cu2+, Fe2+, Ni2+, Mn2+ uncultured microorganism

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.135 66000
-
 SDS-PAGE, 605 amino acid product of 1815 bp gene sequence with N-terminal signal sequence of 27 amino acids (M1-Q27) uncultured microorganism

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.135 uncultured microorganism A7LI67 gene Env Npu193A of environmental soil DNA sample from a hot spring in Thailand and expressed in yeast Pichia pastoris KM71 with plasmid pPICZalphaA
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.135 gene purification by agarose gel electrophoresis, enzyme purification by size exclusion chromatography, elution via Superdex 200 10/300 GL column with 50 mM MOPS, pH7, 50 mM NaCl uncultured microorganism

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.135 additional information
-
 very poor activity with substrate gamma-cyclodextrin, 50 mM MOPS buffer, pH 7.0, 2 mM CaCl2, 75°C, 45 min uncultured microorganism
3.2.1.135 0.15
-
 Vmax, substrate starch, 50 mM MOPS buffer, pH 7.0, 2 mM CaCl2, 75°C, 45 min uncultured microorganism
3.2.1.135 23.24
-
 Vmax, substrate pullulan, 50 mM MOPS buffer, pH 7.0, 2 mM CaCl2, 75°C, 45 min uncultured microorganism

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.135 gamma-cyclodextrin + H2O not alpha or beta-cyclodextrins uncultured microorganism maltose + ? no glucose ?
3.2.1.135 linear maltooligosaccharides + H2O
-
 uncultured microorganism maltose + ? no glucose ?
3.2.1.135 pullulan + H2O
-
 uncultured microorganism maltotriose + ? no panose ?
3.2.1.135 starch + H2O not alpha or beta-cyclodextrins uncultured microorganism maltose + ? no glucose ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.135 cyclomaltodextrinase
-
 uncultured microorganism
3.2.1.135 Env Npu193A
-
 uncultured microorganism
3.2.1.135 neopullulanase-like enzyme
-
 uncultured microorganism

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.135 75
-
 substrate pullulan: 50 mM MOPS, pH 7, 2 mM CaCl2 uncultured microorganism

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.135 30 90 substrate pullulan: more than 80% activity from 65-80°C, 70-80% activity from 55-60°C, about 40% activity between 40 and 50°C, about 20-30% activity at 30°C and beyond 80°C uncultured microorganism

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.135 45 75 substrate pullulan, pre-incubation at 45, 50, 60, 65, 70, 75°C for 30, 60, and 120 min. about 80% activity after 120 min between 45 and 60°C, more than 60% activity after 120 min at 65°C, about 50% after 120 min at 70°C, around 20% activity after 120 min at 75°C uncultured microorganism

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.135 7
-
 substrate pullulan uncultured microorganism

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.135 4 11 substrate pullulan: at least 50% activity retained between pH 5 and 9, about 20% activity at pH 4 and beyond pH 10. buffers of 100 mM pH 4: sodium citrate, pH 5-6: acetic acid, pH 6.5-7: MOPS, pH 7-9: Tris-HCl, pH 9-11: glycine uncultured microorganism

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.135 3 9 substrate pullulan, 25°C, 4 h pre-incubation at pH 3: 50 mM sodium citrate, pH 5: 50 mM sodium acetate, pH 7: 50 mM Tris-HCl, pH 9: 50 mM glycin, more than 50% activity retained at pH 3, more than 80% at pH 5, about 100% at pH 7 and 9 uncultured microorganism