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Literature summary for 2.4.1.100 extracted from

  • Lasseur, B.; Schroeven, L.; Lammens, W.; Le Roy, K.; Spangenberg, G.; Manduzio, H.; Vergauwen, R.; Lothier, J.; Prudhomme, M.P.; Van den Ende, W.
    Transforming a fructan:fructan 6G-fructosyltransferase from perennial ryegrass into a sucrose:sucrose 1-fructosyltransferase (2009), Plant Physiol., 149, 327-339.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Pichia pastoris Lolium perenne

Protein Variants

Protein Variants Comment Organism
N340D the mutant shows only very low overall activity levels Lolium perenne
N340D/S415N the mutant shows similar properties to the W343R mutant, but the 1-nystose/neoketose ratio is decreased Lolium perenne
N340D/W343R the mutant shows 6G-FFT and 1-FFT activities that hold the middle between those of the N340D and W343R single mutants Lolium perenne
N340D/W343R/S415N the triple mutant gains the characteristic of using sucrose as a donor substrate, at the same time, this mutant can still use 1-kestose as a donor substrate, but in a much less efficient way than the wild type Lolium perenne
S415N the mutant shows slightly lower 6G-FFT/1-FFT activities, but the ratio between the main reaction products is not significantly affected Lolium perenne
W343R the mutant shows a wild type1-FFT activity but a largely reduced 6G-FFT activity Lolium perenne

General Stability

General Stability Organism
several attempts to purify 6G-FFT/1-FFT are unsuccessful, probably due to the instability of the enzyme and/or the rather low concentration in the yeast's expression medium Lolium perenne

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
976
-
D-sucrose mutant enzyme N340D/W343R/S415N, in 50 mM sodium acetate buffer, pH 5.0, at 30°C Lolium perenne

Organism

Organism UniProt Comment Textmining
Lolium perenne Q8S337 perennial ryegrass
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate precipitation, several attempts (concanavalin A, Mono Q, Mono S) to further purify 6G-FFT/1-FFT are unsuccessful, probably due to the instability of the enzyme and/or the rather low concentration in the yeast’s expression medium Lolium perenne

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1-kestose + 1-kestose 1-FFT activity Lolium perenne D-sucrose + 1,1-nystose
-
?
additional information a minor 1-SST activity is only found with D-sucrose as a single substrate if there is no 1-kestose available, while 1-kestose hydrolytic activity becomes gradually more important at low concentrations Lolium perenne ?
-
?
additional information the bifunctional 6G-FFT/1-FFT uses 1-kestotriose as a preferential donor substrate and can create both a beta(2->1) linkage between two fructosyl residues and a beta(2->6) linkage between one fructosyl residue and one glucosyl residue Lolium perenne ?
-
?

Synonyms

Synonyms Comment Organism
6G-FFT/1-FFT
-
Lolium perenne
fructan:fructan 6G-fructosyltransferase/fructan:fructan 1-fructosyltransferase
-
Lolium perenne