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Literature summary extracted from

  • Kuehn, A.; Yu, S.; Giffhorn, F.
    Catabolism of 1,5-anhydro-D-fructose in Sinorhizobium morelense S-30.7.5: discovery, characterization, and overexpression of a new 1,5-anhydro-D-fructose reductase and its application in sugar analysis and rare sugar synthesis (2006), Appl. Environ. Microbiol., 72, 1248-1257.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.1.1.263 additional information activity of AFR is not influenced by 1 mM EDTA Ensifer adhaerens

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.263 expressed in Escherichia coli Ensifer adhaerens
1.1.1.292 overexpression in Escherichia coli Ensifer adhaerens

General Stability

EC Number General Stability Organism
1.1.1.263 the highest activity of AFR is measured at 30°C in 100 mM Bistris-HCl at pH 6.5, with 50% remaining activity at pH 5.2 (in citrate buffer) and pH 8.8 (in Tris-HCl buffer) Ensifer adhaerens

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.263 additional information activity of AFR was not influenced by 1 mM EDTA or by any of the given metal salts NaCl, KCl, MgCl2, CaCl2, MnCl2, FeCl3, and ZnCl2, 1 mM each Ensifer adhaerens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.263 0.2
-
 NADPH
-
 Ensifer adhaerens
1.1.1.263 8.4
-
 1,5-Anhydro-D-fructose
-
 Ensifer adhaerens
1.1.1.263 11
-
 D-glucosone
-
 Ensifer adhaerens
1.1.1.292 0.2
-
 NADPH pH 6.5, 30°C Ensifer adhaerens
1.1.1.292 8.4
-
 1,5-Anhydro-D-fructose pH 6.5, 30°C Ensifer adhaerens
1.1.1.292 11
-
 D-glucosone pH 6.5, 30°C Ensifer adhaerens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.263 additional information activity of AFR was not influenced by NaCl, KCl, MgCl2, CaCl2, MnCl2, FeCl3, and ZnCl2, 1 mM each Ensifer adhaerens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.263 35100
-
 MALDI-TOF-MS Ensifer adhaerens
1.1.1.263 38200
-
 gel filtration Ensifer adhaerens
1.1.1.263 40000
-
 SDS-PAGE Ensifer adhaerens
1.1.1.292 35100
-
 1 * 35100, MALDI-TOF-MS Ensifer adhaerens
1.1.1.292 38200
-
 gel filtration Ensifer adhaerens
1.1.1.292 40000
-
 1 * 40000, SDS-PAGE Ensifer adhaerens

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.263 Ensifer adhaerens
-
-
-
1.1.1.263 Ensifer adhaerens S-30.7.5
-
-
-
1.1.1.292 Ensifer adhaerens Q2I8V6 i.e. Ensifer adhaerens
-
1.1.1.292 Ensifer adhaerens S-30.7.5 Q2I8V6 i.e. Ensifer adhaerens
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.263 Q-Sepharose HP column chromatography, Superdex 200 HR10 30 column chromatography and Red Sepharose CL-6B chromatography Ensifer adhaerens
1.1.1.292 native and recombinant enzyme Ensifer adhaerens

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.263 additional information
-
 AFR of S. morelense S-30.7.5 was inactive toward pyridine-3-aldehyde, 2,3-butanedione, glucuronic acid, acetaldehyde, and formaldehyde Ensifer adhaerens
1.1.1.263 0.01
-
 1,5-anhydro-D-glucitol as substrate Ensifer adhaerens
1.1.1.263 0.06
-
 1,5-anhydro-D-mannitol as substrate Ensifer adhaerens
1.1.1.263 6.5
-
 in crude cell extract Ensifer adhaerens
1.1.1.263 21
-
 ascopyrone as substrate Ensifer adhaerens
1.1.1.263 21
-
 D-galactosone as substrate Ensifer adhaerens
1.1.1.263 53
-
 D-allosone as substrate Ensifer adhaerens
1.1.1.263 88
-
 D-xylosone as substrate Ensifer adhaerens
1.1.1.263 118
-
 6-deoxy-D-glucosone as substrate Ensifer adhaerens
1.1.1.263 118
-
 D-glucosone as substrate Ensifer adhaerens
1.1.1.263 191
-
 3-keto-1,5-anhydro-D-fructose as substrate Ensifer adhaerens
1.1.1.263 488.9
-
 after 75 fold purification Ensifer adhaerens
1.1.1.263 530
-
 1,5-anhydro-D-fructose as substrate Ensifer adhaerens
1.1.1.292 288.9
-
 native enzyme Ensifer adhaerens
1.1.1.292 484
-
 recombinant enzyme expressed in Escherichia coli Ensifer adhaerens

Storage Stability

EC Number Storage Stability Organism
1.1.1.263 -20°C, 50 days, 50% loss of activity Ensifer adhaerens
1.1.1.263 0°C, 50 days, 50% loss of activity Ensifer adhaerens
1.1.1.292 -20°C or 0°C, 50 days, 50% loss of activity Ensifer adhaerens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.263 1,5-anhydro-D-fructose + NADP+
-
 Ensifer adhaerens 1,5-anhydro-D-mannitol + NADPH
-
 r
1.1.1.263 1,5-anhydro-D-fructose + NADP+
-
 Ensifer adhaerens S-30.7.5 1,5-anhydro-D-mannitol + NADPH
-
 r
1.1.1.263 1,5-anhydro-D-glucitol + NADPH
-
 Ensifer adhaerens ? + NADP+
-
 ?
1.1.1.263 1,5-anhydro-D-glucitol + NADPH
-
 Ensifer adhaerens S-30.7.5 ? + NADP+
-
 ?
1.1.1.263 1,5-anhydro-D-mannitol + NADPH
-
 Ensifer adhaerens ? + NADP+
-
 ?
1.1.1.263 1,5-anhydro-D-mannitol + NADPH
-
 Ensifer adhaerens S-30.7.5 ? + NADP+
-
 ?
1.1.1.263 3-keto-1,5-anhydro-D-fructose + NADP+
-
 Ensifer adhaerens 1,5-anhydro-D-mannitol + NADPH
-
 ?
1.1.1.263 3-keto-1,5-anhydro-D-fructose + NADP+
-
 Ensifer adhaerens S-30.7.5 1,5-anhydro-D-mannitol + NADPH
-
 ?
1.1.1.263 6-deoxy-D-glucosone + NADP+
-
 Ensifer adhaerens 6-deoxy-D-mannose + NADPH
-
 ?
1.1.1.263 6-deoxy-D-glucosone + NADP+
-
 Ensifer adhaerens S-30.7.5 6-deoxy-D-mannose + NADPH
-
 ?
1.1.1.263 ascopyrone P + NADP+
-
 Ensifer adhaerens ? + NADPH
-
 ?
1.1.1.263 D-allosone + NADP+
-
 Ensifer adhaerens D-altrose + NADPH
-
 ?
1.1.1.263 D-galactosone + NADP+
-
 Ensifer adhaerens D-talose + NADPH
-
 ?
1.1.1.263 D-glucosone + NADP+
-
 Ensifer adhaerens D-mannose + NADPH
-
 ?
1.1.1.263 D-xylosone + NADP+
-
 Ensifer adhaerens D-lyxose + NADPH
-
 ?
1.1.1.292 1,5-anhydro-D-fructose + NADPH + H+
-
 Ensifer adhaerens 1,5-anhydro-D-mannitol + NADP+
-
 r
1.1.1.292 1,5-anhydro-D-fructose + NADPH + H+
-
 Ensifer adhaerens S-30.7.5 1,5-anhydro-D-mannitol + NADP+
-
 r
1.1.1.292 1,5-anhydro-D-glucitol + NADP+ 23% of the activity with 1,5-anhydro-D-mannitol Ensifer adhaerens ?
-
 ?
1.1.1.292 1,5-anhydro-D-glucitol + NADP+ 23% of the activity with 1,5-anhydro-D-mannitol Ensifer adhaerens S-30.7.5 ?
-
 ?
1.1.1.292 6-deoxy-D-glucosone + NADPH + H+ 22% of the activity with 1,5-anhydro-D-fructose Ensifer adhaerens 6-deoxy-D-mannose + NADP+
-
 ?
1.1.1.292 6-deoxy-D-glucosone + NADPH + H+ 22% of the activity with 1,5-anhydro-D-fructose Ensifer adhaerens S-30.7.5 6-deoxy-D-mannose + NADP+
-
 ?
1.1.1.292 D-allosone + NADPH + H+ 10% of the activity with 1,5-anhydro-D-fructose Ensifer adhaerens D-altrose + NADP+
-
 ?
1.1.1.292 D-allosone + NADPH + H+ 10% of the activity with 1,5-anhydro-D-fructose Ensifer adhaerens S-30.7.5 D-altrose + NADP+
-
 ?
1.1.1.292 D-glucosone + NADPH + H+ 22% of the activity with 1,5-anhydro-D-fructose Ensifer adhaerens D-mannose + NADP+
-
 ?
1.1.1.292 D-glucosone + NADPH + H+ 22% of the activity with 1,5-anhydro-D-fructose Ensifer adhaerens S-30.7.5 D-mannose + NADP+
-
 ?
1.1.1.292 D-xylosone + NADPH + H+ 17% of the activity with 1,5-anhydro-D-fructose Ensifer adhaerens D-lyxose + NADP+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.263 monomer 1 * 35100, MALDI-TOF-MS Ensifer adhaerens
1.1.1.292 monomer 1 * 40000, SDS-PAGE Ensifer adhaerens
1.1.1.292 monomer 1 * 35100, MALDI-TOF-MS Ensifer adhaerens

Synonyms

EC Number Synonyms Comment Organism
1.1.1.263 AFR
-
 Ensifer adhaerens
1.1.1.263 NADPH-dependent monomeric reductase
-
 Ensifer adhaerens
1.1.1.292 1,5-anhydro-D-fructose reductase (ambiguous)
-
 Ensifer adhaerens
1.1.1.292 AFR
-
 Ensifer adhaerens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.263 63.2
-
 D-glucosone
-
 Ensifer adhaerens
1.1.1.263 286.7
-
 1,5-Anhydro-D-fructose
-
 Ensifer adhaerens
1.1.1.263 1300
-
 1,5-Anhydro-D-fructose
-
 Ensifer adhaerens
1.1.1.292 63.2
-
 D-glucosone pH 6.5, 30°C Ensifer adhaerens
1.1.1.292 286.7
-
 1,5-Anhydro-D-fructose pH 6.5, 30°C Ensifer adhaerens
1.1.1.292 1300
-
 1,5-Anhydro-D-fructose pH 6.5, 30°C Ensifer adhaerens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.292 6.5
-
-
 Ensifer adhaerens

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.292 5.2 8.8 about 50% of maximal activity at pH 5.5 and at pH 8.8 Ensifer adhaerens

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.1.1.263 5.2 8.8 at 30°C the highest activity of AFR is measured in 100 mM Bistris-HCl at pH 6.5, with 50% remaining activity at pH 5.2 (in citrate buffer) and pH 8.8 (in Tris-HCl buffer) Ensifer adhaerens

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.263 NADPH
-
 Ensifer adhaerens
1.1.1.292 NADP+
-
 Ensifer adhaerens
1.1.1.292 NADPH inactive towards NADH Ensifer adhaerens

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.1.1.263 Ensifer adhaerens
-
-
 4.3
1.1.1.292 Ensifer adhaerens isoelectric focusing
-
 4.3