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Literature summary extracted from

  • Nishioka, T.; Yasutake, Y.; Nishiya, Y.; Tamura, T.
    Structure-guided mutagenesis for the improvement of substrate specificity of Bacillus megaterium glucose 1-dehydrogenase IV (2012), FEBS J., 279, 3264-3275.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.47 expressed as a His-tagged fusion protein Priestia megaterium

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.47 the crystal structures of BmGlcDH-IV in ligand-free, NADH-bound and beta-D-glucose-bound forms is determined to a resolution of 2.0 A Priestia megaterium

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.47 A258F Km (D-glucose) increased at pH 6 compared to wild-type, decreased at pH 8 compared to wild-type. Mutant shows higher substrate specificity with D-xylose, D-mannose, D-galactose and D-glucosamine compared to wild-type. Mutant shows a markedly deteriorated thermostability Priestia megaterium
1.1.1.47 DELTAG261 Km (D-glucose) highly increased at pH 6 and pH 8 compared to wild-type.Specific activity of mutant for D-glucose is severely decreased at both pH 6.0 and pH 8.0. Mutant shows a markedly deteriorated thermostability Priestia megaterium
1.1.1.47 G259A Km (D-glucose) increased at pH 6 and pH 8 compared to wild-type. Specific activity of the G259A mutant is slightly lower, but is still comparable with wild-type BmGlcDH-IV. Thermostability comparable to wild-type Priestia megaterium
1.1.1.47 G259V Km (D-glucose) highly increased at pH 6 and pH 8 compared to wild-type. Specific activity of mutant for D-glucose is severely decreased at both pH 6.0 and pH 8.0. Thermostability comparable to wild-type Priestia megaterium
1.1.1.47 G261A Km (D-glucose) highly increased at pH 6 and pH 8 compared to wild-type. Specific activity of mutant for D-glucose is severely decreased at both pH 6.0 and pH 8.0. Thermostability comparable to wild-type Priestia megaterium
1.1.1.47 G261V Km (D-glucose) highly increased at pH 6 and pH 8 compared to wild-type. Specific activity of mutant for D-glucose is severely decreased at both pH 6.0 and pH 8.0. Thermostability comparable to wild-type Priestia megaterium

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.47 3.1
-
 D-glucose pH 6.0, 37°C, wild-tpye Priestia megaterium
1.1.1.47 4 10 D-glucose pH 6.0, 37°C, mutant G261A Priestia megaterium
1.1.1.47 4.3
-
 D-glucose pH 6.0, 37°C, mutant A258F Priestia megaterium
1.1.1.47 9.8
-
 D-glucose pH 8.0, 37°C, mutant A258F Priestia megaterium
1.1.1.47 11
-
 D-glucose pH 6.0, 37°C, wild-tpye Priestia megaterium
1.1.1.47 12
-
 D-glucose pH 8.0, 37°C, wild-tpye Priestia megaterium
1.1.1.47 63
-
 D-glucose pH 8.0, 37°C, wild-tpye Priestia megaterium
1.1.1.47 81
-
 D-glucose pH 6.0, 37°C, mutant G259A Priestia megaterium
1.1.1.47 380
-
 D-glucose pH 8.0, 37°C, mutant G259A Priestia megaterium
1.1.1.47 1100
-
 D-glucose pH 6.0, 37°C, mutant DELTAG261 Priestia megaterium
1.1.1.47 1400
-
 D-glucose pH 6.0, 37°C, mutant G259V Priestia megaterium
1.1.1.47 2000
-
 D-glucose pH 8.0, 37°C, mutant DELTAG261 Priestia megaterium
1.1.1.47 2000
-
 D-glucose pH 8.0, 37°C, mutant G261A Priestia megaterium
1.1.1.47 4000
-
 D-glucose pH 6.0, 37°C, mutant G261V Priestia megaterium
1.1.1.47 4000
-
 D-glucose pH 8.0, 37°C, mutant G259V Priestia megaterium
1.1.1.47 5000
-
 D-glucose pH 8.0, 37°C, mutant G261V Priestia megaterium

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.47 Priestia megaterium
-
-
-
1.1.1.47 Priestia megaterium P39485
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.47 D-galactose + NAD+ substrate specificity wild-tpye: 2.9% (reference: D-glucose 100%) Priestia megaterium D-galactono-1,5-lactone + NADH + H+
-
 ?
1.1.1.47 D-galactose + NAD+ substrate specificity wild-tpye: 9.1% (reference: D-glucose 100%) Priestia megaterium D-galactono-1,5-lactone + NADH + H+
-
 ?
1.1.1.47 D-glucosamine + NAD+ substrate specificity wild-tpye: 11% (reference: D-glucose 100%) Priestia megaterium D-glucosamino-1,5-lactone + NADH + H+
-
 ?
1.1.1.47 D-glucosamine + NAD+ substrate specificity wild-tpye: 3% (reference: D-glucose 100%) Priestia megaterium D-glucosamino-1,5-lactone + NADH + H+
-
 ?
1.1.1.47 D-glucose + NAD+
-
 Priestia megaterium D-glucono-1,5-lactone + NADH + H+
-
 ?
1.1.1.47 D-mannose + NAD+ substrate specificity wild-tpye: 18% (reference: D-glucose 100%) Priestia megaterium D-mannono-1,5-lactone + NADH + H+
-
 ?
1.1.1.47 D-mannose + NAD+ substrate specificity wild-tpye: 4.8% (reference: D-glucose 100%) Priestia megaterium D-mannono-1,5-lactone + NADH + H+
-
 ?
1.1.1.47 D-xylose + NAD+ substrate specificity wild-tpye: 10% (reference: D-glucose 100%) Priestia megaterium D-xylono-1,5-lactone + NADH + H+
-
 ?
1.1.1.47 D-xylose + NAD+ substrate specificity wild-tpye: 35% (reference: D-glucose 100%) Priestia megaterium D-xylono-1,5-lactone + NADH + H+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.1.47 BmGlcDH-III
-
 Priestia megaterium
1.1.1.47 BmGlcDH-IV
-
 Priestia megaterium
1.1.1.47 glucose 1-dehydrogenase III
-
 Priestia megaterium
1.1.1.47 glucose 1-dehydrogenase IV
-
 Priestia megaterium

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.47 37
-
 assay at Priestia megaterium

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.47 65
-
 wild-type enzyme retains 20% activity after 20 min at 65°C Priestia megaterium
1.1.1.47 65
-
 wild-type enzyme retains 70% activity after 40 min at 65°C Priestia megaterium

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.47 8
-
 assay at Priestia megaterium