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Literature summary extracted from

  • Ferrer, M.; Golyshina, O.V.; Plou, F.J.; Timmis, K.N.; Golyshin, P.N.
    A novel alpha-glucosidase from the acidophilic archaeon Ferroplasma acidiphilum strain Y with high transglycosylation activity and an unusual catalytic nucleophile (2005), Biochem. J., 391, 269-276.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.20 expression in Escherichia coli Ferroplasma acidiphilum

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.20 H243A kcat/Km is 23% lower than wild-type value, the iron content is about 55% compared to wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 H275A kcat/Km is slighly higher than wild-type value, the iron content is similar to that of the wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 H277A kcat/Km is slighly lower than wild-type value, the iron content is similar to that of the wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 H325A kcat/Km is slighly higher than wild-type value, the iron content is similar to that of the wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 H390A kcat/Km is 98% lower than wild-type value, contains no iron Ferroplasma acidiphilum
3.2.1.20 H41A kcat/Km is slighly higher than wild-type value, the iron content is similar to that of the wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 H47A kcat/Km is 25% lower than wild-type value, the iron content is about 85% compared to wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 H9A kcat/Km is slighly higher than wild-type value, the iron content is similar to that of the wild-type enzyme Ferroplasma acidiphilum

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.20 5-fluoro-alpha-D-glucopyranosyl fluoride strong inhibition by the reversible inhibitor, which inhibits immediately upon addition and maintains the same level of inhibition over time Ferroplasma acidiphilum
3.2.1.20 diethyl dicarbonate
-
 Ferroplasma acidiphilum
3.2.1.20 additional information no inactivation with 1-ethyl-3-[3-(dimethylamino)propyl]carbodi-imide, tetranitromethane, N-ethylmaleimide, iodoacetate, p-chloromercuribenzoate, 4-chloromercuriphenylsulfonic acid, di-isopropyl fluorophosphate, PMSF. The presence of glucose and maltose, the end-products of the hydrolysis of sucrose, maltose and maltotriose, does not affect the hydrolytic activity of the enzyme Ferroplasma acidiphilum
3.2.1.20 tosylphenylalanylchloromethane
-
 Ferroplasma acidiphilum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.20 0.32
-
 sucrose pH 3.0, 50°C, wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 0.64
-
 maltose pH 3.0, 50°C, wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 0.69
-
 maltotriose pH 3.0, 50°C, wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 5
-
 4-nitrophenyl alpha-D-glucopyranoside pH 3.0, 50°C, wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 9.3
-
 maltotetraose pH 3.0, 50°C, wild-type enzyme Ferroplasma acidiphilum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.20 Iron the enzyme contains 1.04 mol of iron per mol of enzyme Ferroplasma acidiphilum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.20 57000
-
 x * 57000, SDS-PAGE Ferroplasma acidiphilum
3.2.1.20 57300
-
 x * 57300, calculated from sequence Ferroplasma acidiphilum

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.20 Ferroplasma acidiphilum Q5K3Q3
-
-
3.2.1.20 Ferroplasma acidiphilum DSM 12658 Q5K3Q3
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.20 4-nitrophenyl alpha-D-glucopyranoside + H2O
-
 Ferroplasma acidiphilum 4-nitrophenol + alpha-D-glucose
-
 ?
3.2.1.20 maltose + H2O
-
 Ferroplasma acidiphilum 2 D-glucose
-
 ?
3.2.1.20 maltose + H2O
-
 Ferroplasma acidiphilum DSM 12658 2 D-glucose
-
 ?
3.2.1.20 maltotetraose + H2O very low activity Ferroplasma acidiphilum ?
-
 ?
3.2.1.20 maltotetraose + H2O very low activity Ferroplasma acidiphilum DSM 12658 ?
-
 ?
3.2.1.20 maltotriose + H2O
-
 Ferroplasma acidiphilum ?
-
 ?
3.2.1.20 maltotriose + H2O
-
 Ferroplasma acidiphilum DSM 12658 ?
-
 ?
3.2.1.20 additional information the enzyme also shows high transglycosylation activity, it transfers glucosyl groups from maltose to produce maltotriose selectively Ferroplasma acidiphilum ?
-
 ?
3.2.1.20 additional information the enzyme also shows high transglycosylation activity, it transfers glucosyl groups from maltose to produce maltotriose selectively Ferroplasma acidiphilum DSM 12658 ?
-
 ?
3.2.1.20 sucrose + H2O
-
 Ferroplasma acidiphilum D-glucose + D-fructose
-
 ?
3.2.1.20 sucrose + H2O
-
 Ferroplasma acidiphilum DSM 12658 D-glucose + D-fructose
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.20 ? x * 57000, SDS-PAGE Ferroplasma acidiphilum
3.2.1.20 ? x * 57300, calculated from sequence Ferroplasma acidiphilum

Synonyms

EC Number Synonyms Comment Organism
3.2.1.20 alphaGluFa
-
 Ferroplasma acidiphilum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.20 50
-
 assay at Ferroplasma acidiphilum
3.2.1.20 55 60 hydrolysis of sucrose Ferroplasma acidiphilum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.20 50
-
 pH 2.5, t1/2: 34 min Ferroplasma acidiphilum
3.2.1.20 55
-
 thermostability: 57 min Ferroplasma acidiphilum
3.2.1.20 60
-
 thermostability: 32 vmin Ferroplasma acidiphilum
3.2.1.20 65
-
 thermostability: less than 5 min Ferroplasma acidiphilum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.20 2.8
-
 maltotetraose pH 3.0, 50°C, wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 13
-
 maltotriose pH 3.0, 50°C, wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 22.5
-
 4-nitrophenyl alpha-D-glucopyranoside pH 3.0, 50°C, wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 94
-
 sucrose pH 3.0, 50°C, wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 126
-
 maltose pH 3.0, 50°C, wild-type enzyme Ferroplasma acidiphilum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.20 2.4 3.5 hydrolysis of sucrose Ferroplasma acidiphilum
3.2.1.20 3
-
 assay at Ferroplasma acidiphilum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.20 1.5 6 pH 1.5: 74% of maximal activity, pH 2.0: 92% of maximal activity, pH 6.0: about 35% of maximal activity. No measurable activity is detected at pH values above 7.0 and below 1.0 Ferroplasma acidiphilum

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.20 2.5
-
 50°C, t1/2: 34 min Ferroplasma acidiphilum

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.2.1.20 0.0029
-
 diethyl dicarbonate pH 3.0, 50°C Ferroplasma acidiphilum
3.2.1.20 0.0042
-
 tosylphenylalanylchloromethane pH 3.0, 50°C Ferroplasma acidiphilum

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.2.1.20 Ferroplasma acidiphilum calculated from sequence
-
 6.42

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.20 0.3
-
 maltotetraose pH 3.0, 50°C, wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 4.5
-
 4-nitrophenyl alpha-D-glucopyranoside pH 3.0, 50°C, wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 5.4
-
 sucrose pH 3.0, 50°C, mutant enzyme H390A Ferroplasma acidiphilum
3.2.1.20 18.8
-
 maltotriose pH 3.0, 50°C, wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 197
-
 maltose pH 3.0, 50°C, wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 220
-
 sucrose pH 3.0, 50°C, mutant enzyme H47A Ferroplasma acidiphilum
3.2.1.20 227
-
 sucrose pH 3.0, 50°C, mutant enzyme H243A Ferroplasma acidiphilum
3.2.1.20 291
-
 sucrose pH 3.0, 50°C, mutant enzyme H277A Ferroplasma acidiphilum
3.2.1.20 293
-
 sucrose pH 3.0, 50°C, wild-type enzyme Ferroplasma acidiphilum
3.2.1.20 296
-
 sucrose pH 3.0, 50°C, mutant enzyme H325A Ferroplasma acidiphilum
3.2.1.20 297
-
 sucrose pH 3.0, 50°C, mutant enzyme H275A Ferroplasma acidiphilum
3.2.1.20 302
-
 sucrose pH 3.0, 50°C, mutant enzyme H9A Ferroplasma acidiphilum
3.2.1.20 312
-
 sucrose pH 3.0, 50°C, mutant enzyme H41A Ferroplasma acidiphilum