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Literature summary extracted from

  • Le Roy, K.; Lammens, W.; Van Laere, A.; Van den Ende, W.
    Influencing the binding configuration of sucrose in the active sites of chicory fructan 1-exohydrolase and sugar beet fructan 6-exohydrolase (2008), New Phytol., 178, 572-580.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.80
-
 Cichorium intybus
3.2.1.153 heterologous expression in Pichia pastoris, pPICZalphaA vector Cichorium intybus
3.2.1.154 heterologous expression in Pichia pastoris, pPICZalphaA vector Beta vulgaris

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.80 S101L strongly increased sucrose-hydrolysing activity compared with the wild-type enzym at high sucrose concentrations. At a very low substrate concentration (5 mm), the introduced sucrose hydrolysing activity is small compared with the activities towards the preferential 1-FEH IIa substrates 1-kestose and inulin Cichorium intybus
3.2.1.153 S101L neighbouring amino acid exchange influences the orientation of Trp82, thus switches the function of sucrose as inhibitor towards hydrolysation substrate, but only when present in high concentrations because substrate binding is inefficient Cichorium intybus
3.2.1.154 F233D introduction of acidic amino acid in vicinity of acid-base catalyst creating a beta-fructofuranosidase type of enzyme with dual activity against sucrose and levan, generally lower activities against levan and 6-kestose but conserving preference for these substrates over sucrose Beta vulgaris

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.153 sucrose inhibitor for wild-type 1-FEH IIa, weak hydrolysation product in S101L mutant Cichorium intybus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.153 additional information Cichorium intybus hydrolytic cleavage of terminal fructosyl residues off inulin ?
-
 ?
3.2.1.154 additional information Beta vulgaris hydrolytic cleavage of terminal fructosyl residues off inulin ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.80 Cichorium intybus Q93X60 precursor
-
3.2.1.153 Cichorium intybus
-
-
-
3.2.1.154 Beta vulgaris Q70XE6
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.153
-
 Cichorium intybus
3.2.1.154
-
 Beta vulgaris

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.80 1-kestose + H2O
-
 Cichorium intybus beta-D-fructose + sucrose
-
 ?
3.2.1.80 inulin + H2O
-
 Cichorium intybus ?
-
 ?
3.2.1.153 1-kestose + H2O
-
 Cichorium intybus ?
-
 ?
3.2.1.153 inulin + H2O 5% inulin from chicory, 30 degrees Celsius Cichorium intybus fructose + fructo-oligosaccharide + fructo-polysaccharide
-
 ?
3.2.1.153 additional information hydrolytic cleavage of terminal fructosyl residues off inulin Cichorium intybus ?
-
 ?
3.2.1.153 sucrose + H2O
-
 Cichorium intybus D-fructose + D-glucose
-
 ?
3.2.1.154 6-kestose + H2O
-
 Beta vulgaris ?
-
 ?
3.2.1.154 levan + H2O
-
 Beta vulgaris ?
-
 ?
3.2.1.154 levansucrase + H2O Bacillus subtilis Beta vulgaris ?
-
 ?
3.2.1.154 additional information hydrolytic cleavage of terminal fructosyl residues off inulin Beta vulgaris ?
-
 ?
3.2.1.154 sucrose + H2O
-
 Beta vulgaris D-fructose + D-glucose
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.80 Ci1-1-FEH IIa
-
 Cichorium intybus
3.2.1.80 fructan 1-exohydrolase
-
 Cichorium intybus
3.2.1.153 1-FEH I
-
 Cichorium intybus
3.2.1.153 1-FEH IIa
-
 Cichorium intybus
3.2.1.153 fructan 1-exohydrolase
-
 Cichorium intybus
3.2.1.154 6-FEH
-
 Beta vulgaris
3.2.1.154 fructan 6-exohydrolase
-
 Beta vulgaris

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.153 additional information
-
 sucrose 5 mM substrate: about zero activity in both enzyme variants, only at higher substrate concentrations (500-1000 mM) strongly raised activity: 10 mol fructose/mol enzyme Cichorium intybus
3.2.1.153 additional information
-
 inulin 5 mM substrate: wild-type 1-FEH IIa 10 mol fructose/mol enzyme, S101L mutant about 2 mol fructose/mol enzyme Cichorium intybus
3.2.1.153 additional information
-
 1-kestose 5 mM substrate: wild-type enzyme about 23 mol fructose/mol enzyme, mutant about 8 mol fructose/mol enzyme Cichorium intybus
3.2.1.154 additional information
-
 sucrose 5 mM substrate: about zero acitivity in wild-type, mutant about 0.1 mol fructose/mol enzyme, at 1000 mM strongly raised activity: 4 mol fructose/mol enzyme Beta vulgaris
3.2.1.154 additional information
-
 levan 5 mM substrate: wild-type 6-FEH about 2.2 mol fructose/mol enzyme, F233D mutant about 0.4 mol fructose/mol enzyme Beta vulgaris
3.2.1.154 additional information
-
 6-kestose 5 mM substrate: wild-type enzyme about 1.8 mol fructose/mol enzyme, mutant about 0.3 mol fructose/mol enzyme Beta vulgaris