Purification, cloning, and functional expression of sucrose:fructan 6-fructosyltransferase, a key enzyme of fructan synthesis in barley.
- 5 December 1995
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 92 (25), 11652-11656
- https://doi.org/10.1073/pnas.92.25.11652
Abstract
Fructans play an important role in assimilate partitioning and possibly in stress tolerance in many plant families. Sucrose:fructan 6-fructosyltransferase (6-SFT), an enzyme catalyzing the formation and extension of beta-2,6-linked fructans typical of grasses, was purified from barley (Hordeum vulgare L.). It occurred in two closely similar isoforms with indistinguishable catalytic properties, both consisting of two subunits with apparent masses of 49 and 23 kDa. Oligonucleotides, designed according to the sequences of tryptic peptides from the large subunit, were used to amplify corresponding sequences from barley cDNA. The main fragment generated was cloned and used to screen a barley cDNA expression library. The longest cDNA obtained was transiently expressed in Nicotiana plumbaginifolia protoplasts and shown to encode a functional 6-SFT. The deduced amino acid sequence of the cDNA comprises both subunits of 6-SFT. It has high similarity to plant invertases and other beta-fructosyl hydrolases but only little to bacterial fructosyltransferases catalyzing the same type of reaction as 6-SFT.Keywords
This publication has 23 references indexed in Scilit:
- cDNA Cloning of Carrot (Daucus carota) Soluble Acid [beta]-Fructofuranosidases and Comparison with the Cell Wall IsoenzymePlant Physiology, 1994
- Purification and characterisation of an Aspergillus niger invertase and its DNA sequenceCurrent Genetics, 1993
- Purification and Characterization of Three Soluble Invertases from Barley (Hordeum vulgare L.) LeavesPlant Physiology, 1993
- A short C-terminal sequence is necessary and sufficient for the targeting of chitinases to the plant vacuole.Proceedings of the National Academy of Sciences, 1991
- Regulation of Sucrose-Sucrose-Fructosyltransferase in Barley LeavesPlant Physiology, 1991
- Cloning and sequencing of the inulinase gene of Kluyveromyces marxianus var. marxianus ATCC 12424FEBS Letters, 1991
- cDNA cloning of carrot extracellular beta-fructosidase and its expression in response to wounding and bacterial infection.Plant Cell, 1990
- Regulation of Fructan Metabolism in Leaves of Barley (Hordeum vulgare L. cv Gerbel)Plant Physiology, 1986
- The DNA sequence of the gene for the secreted Bacillus subtilis enzyme levansucrase and its genetic control sitesMolecular Genetics and Genomics, 1985
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970