Fructans (polyfructosylsucrose) are synthesized by a number of plants
and micro-organisms. Plant fructans are
localized in the vacuole and have a low degree of polymerization (DP), whereas
the fructans synthesized by micro-organisms are usually much bigger. There is
an increasing interest in fructans for both food and non-food
applications. In order to accumulate fructans of high DP in the plant vacuole,
the levansucrase protein of Bacillus
subtilis was fused to the vacuolar targeting sequence of sporamin and expressed
in plants. Transgenic tobacco
plants in which this fusion gene is expressed accumulate fructans to levels
up to 21% of the d. wt. They showed
a reduced translocation of carbohydrates, bleaching of the leaves, stunted
growth and increased levels of hexoses
and starch. The levansucrase protein was not translocated to the plant vacuole,
but retained in the endomembrane
system, even though the same targeting signal was able to translocate the E.
coli GUS protein to the plant vacuole.