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Characterisation of disproportionating enzyme from wheat endosperm

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Abstract

Disproportionating enzyme or D-enzyme (EC 2.4.1.25) is an α-1,4 glucanotransferase which catalyses cleavage and transfer reactions involving α-1,4 linked glucans altering (disproportionating) the chain length distribution of pools of oligosaccharides. While D-enzyme has been well characterised in some plants, e.g. potato and Arabidopsis, very little is known about its abundance and function in cereals which constitute the major source of starch worldwide. To address this we have investigated D-enzyme in wheat (Triticum aestivum). Two putative D-enzyme cDNA clones have been isolated from tissue-specific cDNA libraries. TaDPE1-e, from an endosperm cDNA library, encodes a putative polypeptide of 575 amino acid residues including a predicted transit peptide of 41 amino acids. The second cDNA clone, TaDPE1-l, from an Aegilops taushii leaf cDNA library, encodes a putative polypeptide of 579 amino acids including a predicted transit peptide of 45 amino acids. The mature polypeptides TaDPE1-e and TaDPE1-l were calculated to be 59 and 60 kDa, respectively, and had 96% identity. The putative polypeptides had significant identity with deduced D-enzyme sequences from corn and rice, and all the expected conserved residues were present. Protein analysis revealed that D-enzyme is present in the amyloplast of developing endosperm and in the germinating seeds. D-enzyme was partially purified from wheat endosperm and shown to exhibit disproportionating activity in vitro by cleaving maltotriose to produce glucose as well as being able to use maltoheptaose as the donor for the addition of glucans to the outer chains of glycogen and amylopectin.

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Abbreviations

DP:

Degree of polymerisation

dpa:

Days post anthesis

GBSS:

Granule bound starch synthase

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Acknowledgements

NB was the recipient of a postgraduate scholarship from Biogemma. Dr Kevin Gale, CSIRO Plant Industry, is thanked for assistance with design of the synthetic peptide, and Malcolm Blundell for assistance with generation of the antibodies.

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  1. Nicole S. Bresolin & Ian J. Tetlow

    Present address: University of Guelph, N1G 2W1, Guelph, ON, Canada

Authors and Affiliations

  1. CSIRO Plant Industry, GPO Box 1600, Canberra, ACT, 2601, Australia

    Nicole S. Bresolin, Zhongyi Li, Behjat Kosar-Hashemi, Sadequr Rahman, Matthew K. Morell & Crispin A. Howitt

  2. Biogemma UK Ltd, 200 Cambridge Science Park, CB4 0GZ, Cambridge, UK

    Manash Chatterjee

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  1. Nicole S. Bresolin
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Correspondence to Crispin A. Howitt.

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Bresolin, N.S., Li, Z., Kosar-Hashemi, B. et al. Characterisation of disproportionating enzyme from wheat endosperm. Planta 224, 20–31 (2006). https://doi.org/10.1007/s00425-005-0187-7

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