Extraction and characterization of the enzymes of fructan biosynthesis in timothy (Phleum pratense)

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Abstract
A preparation of phlein sucrase from seedling shoots of timothy (Phleum pratense L.) is described which catalyzed the synthesis of fructan with a mean molecular size of 34 000 using sucrose as the substrate. Activity was fully sedimentable at 25 000 × g, had a pH optimum of 7.0, and a Km for sucrose of 0.15 M. Activity was inhibited by β-mercaptoethanol and sodium diethyl dithiocarbamate. Raffinose and stachyose, but not members of the kestose series of oligofructans, could act as fructosyl donors in addition to sucrose. Formation of oligosaccharides during high molecular weight fructan synthesis was minimal, with synthesis occurring by a mechanism apparently analogous to bacterial levansucrase. These observations are discussed in relation to the in vivo patterns of fructan biosynthesis observed in different species of higher plants.