Properties of fructan:fructan 1-fructosyltransferases from chicory and globe thistle, two Asteracean plants storing greatly different types of inulin.
Remarkably, within the Asteraceae, a species-specific fructan pattern can be observed. Some species such as artichoke (Cynara scolymus) and globe thistle (Echinops ritro) store fructans with a considerably higher degree of polymerization than the one observed in chicory (Cichorium intybus) and Jerusalem artichoke (Helianthus tuberosus). Fructan:fructan 1-fructosyltransferase (1-FFT) is the enzyme responsible for chain elongation of inulin-type fructans. 1-FFTs were purified from chicory and globe thistle. A comparison revealed that chicory 1-FFT has a high affinity for sucrose (Suc), fructose (Fru), and 1-kestose as acceptor substrate. This makes redistribution of Fru moieties from large to small fructans very likely during the period of active fructan synthesis in the root when import and concentration of Suc can be expected to be high. In globe thistle, this problem is avoided by the very low affinity of 1-FFT for Suc, Fru, and 1-kestose and the higher affinity for inulin as acceptor substrate. Therefore, the 1-kestose formed by Suc:Suc 1-fructosyltransferase is preferentially used for elongation of inulin molecules, explaining why inulins with a much higher degree of polymerization accumulate in roots of globe thistle. Inulin patterns obtained in vitro from 1-kestose and the purified 1-FFTs from both species closely resemble the in vivo inulin patterns. Therefore, we conclude that the species-specific fructan pattern within the Asteraceae can be explained by the different characteristics of their respective 1-FFTs. Although 1-FFT and bacterial levansucrases clearly differ in their ability to use Suc as a donor substrate, a kinetic analysis suggests that 1-FFT also works via a ping-pong mechanism.
Vergauwen
Rudy
R
K.U. Leuven, Laboratory for Developmental Biology, 3001 Leuven, Belgium.
Van Laere
André
A
Van den Ende
Wim
W
D019659:Asteraceae / Q000201:enzymology; D018651:Cichorium intybus / Q000201:enzymology* / Q000378:metabolism; D031182:Echinops Plant / Q000201:enzymology* / Q000378:metabolism; D005630:Fructans / Q000096:biosynthesis; D005632:Fructose / Q000378:metabolism; D006602:Hexosyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D007444:Inulin / Q000096:biosynthesis*; D007700:Kinetics; D008954:Models, Biological; D010940:Plant Proteins; D013379:Substrate Specificity; D013395:Sucrose / Q000378:metabolism; D014312:Trisaccharides / Q000378:metabolism; D001426:Bacterial Proteins
D005630:Fructans; D005632:Fructose; D006602:Hexosyltransferases; D007444:Inulin; D010940:Plant Proteins; D013395:Sucrose; D014312:Trisaccharides; C072433:1-kestose; C105700:fructan - fructan 6G-fructosyltransferase; C021451:inulosucrase; D001426:Bacterial Proteins