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Literature summary extracted from

  • Dou, S.; Chi, N.; Zhou, X.; Zhang, Q.; Pang, F.; Xiu, Z.
    Molecular cloning, expression, and biochemical characterization of a novel cold-active alpha-amylase from Bacillus sp. dsh19-1 (2018), Extremophiles, 22, 739-749 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.1 DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21T1R(DE3) with molecular chaperones plasmid pGro7 required for enzyme solubility, subcloning in Escherichia coli strain JM109 Bacillus sp. dsh19-1

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.1 Co2+ 27% inhibition at 1 mM, 68% at 10 mM Bacillus sp. dsh19-1
3.2.1.1 Cu2+ 18% inhibition at 1 mM, 48% at 10 mM Bacillus sp. dsh19-1
3.2.1.1 EDTA 41% inhibition at 1 mM, 79% at 10 mM Bacillus sp. dsh19-1
3.2.1.1 Fe3+ 30% inhibition at 1 mM, 62% at 10 mM Bacillus sp. dsh19-1
3.2.1.1 Mn2+ 14% inhibition at 1 mM, 39% at 10 mM Bacillus sp. dsh19-1
3.2.1.1 NaCl enzyme AmyD-1 displays extreme salt tolerance, with the highest activity in the presence of 2.0 M NaCl and 60.5% of activity in 5.0 M NaCl, 40% inhibition at 5 M Bacillus sp. dsh19-1
3.2.1.1 Triton X-100 17% inhibition at 1 mM, 51.5% at 10 mM Bacillus sp. dsh19-1

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.1 additional information
-
additional information Michaelis-Menten kinetics, Km and Vmax of AmyD-1 in 2.0 M NaCl are 2.8 mg/ml and 21.8 mg*ml/ml, respectively, at 20°C and pH 6.0 Bacillus sp. dsh19-1

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.1 Ca2+ activates 9% at 1 mM, 59% at 10 mM Bacillus sp. dsh19-1
3.2.1.1 additional information no or poor effects by Mg2+, K+, and Zn2+ at 1-10 mM Bacillus sp. dsh19-1
3.2.1.1 Na+ activates 15% at 1 mM, 61% at 10 mM Bacillus sp. dsh19-1
3.2.1.1 NaCl enzyme AmyD-1 displays extreme salt tolerance, with the highest activity in the presence of 2.0 M NaCl and 60.5% of activity in 5.0 M NaCl, 40% inhibition at 5 M Bacillus sp. dsh19-1

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.1 Bacillus sp. dsh19-1 A0A192GPZ8
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.1 recombinant His-tagged enzyme 8.2fold from Escherichia coli strain BL21T1R(DE3) by affinity chromatography Bacillus sp. dsh19-1

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 16.4
-
purified recombinant His-tagged enzyme, pH 6.0, 20°C, substrate soluble starch Bacillus sp. dsh19-1

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.1 soluble starch + H2O
-
Bacillus sp. dsh19-1 D-glucose + maltose + maltotriose + maltotetraose
-
?

Subunits

EC Number Subunits Comment Organism
3.2.1.1 ? x * 50100, about, SDS-PAGE and sequence calculation Bacillus sp. dsh19-1

Synonyms

EC Number Synonyms Comment Organism
3.2.1.1 AmyD-1
-
Bacillus sp. dsh19-1
3.2.1.1 cold-active alpha-amylase
-
Bacillus sp. dsh19-1

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.1 20
-
recombinant enzyme Bacillus sp. dsh19-1

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.1 4 70 about 35.7% of maximal activity at 4°C, about 50% at 10°C and 45°C, about 10% at 65°C, inactivation at 75°C Bacillus sp. dsh19-1

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.1 10 45 purified recombinant His-tagged enzyme, over 50% activity remaining, completely stable at 20°C, over 80% activity remaining at 15-35°C Bacillus sp. dsh19-1

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.1 933.5
-
soluble starch recombinant enzyme, 20°C, pH 6.0 Bacillus sp. dsh19-1

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.1 6
-
recombinant enzyme Bacillus sp. dsh19-1

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.1 4 10 over 50% of maximal activity within this range, profile overview Bacillus sp. dsh19-1

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.1 4 9 purified recombinant His-tagged enzyme, over 50% activity remaining, completely stable at pH 6.0, over 80% activity remaining at pH 5.0-8.0 Bacillus sp. dsh19-1