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A xyloglucan-oligosaccharide-specific α-d-xylosidase or exo-oligoxyloglucan-α-xylohydrolase from germinated nasturtium (Tropaeolum majus L.) seeds

Purification, properties and its interaction with a xyloglucan-specific eneto-(1→4)-β-d-glucanase and other hydrolases during storage-xyloglucan mobilisation

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Abstract

The α-xylosidase which is involved in the postgerminative mobilisation of xyloglucan in nasturtium seed cotyledons has now been purified to apparent homogeneity by a facile procedure involving lectin affinity chromatography. The purified enzyme, a glycoprotein, moved as a single band (apparent molecular weight 85000) on sodium dodecyl sulphate-gel electrophoresis, whilst isoelectric focusing gave a number of enzymatically active protein bands spanning the range pI = 5.0 to 7.1 (maximum activity at pI = 6.1). The enzyme did not hydrolyse the simple α-xylosides p-nitrophenyl-α-d-xylopyranoside and woprimeverose (α-d-Xyl(1→6)-d-Glc), or polymeric tamarind-seed xyloglucan. It released xylose from a complex mixture of oligosaccharides produced by exhaustive hydrolysis of tamarind seed xyloglucan using the xyloglucan-specific endo-(1→4)-β-d-glucanase from germinated nasturtium seeds (M. Edwards et al. 1986, J. Biol. Chem., 261. 9489–9494). The three xyloglucan oligosaccharides of lowest molecular size were purified from this mixture and were shown by 1H-nuclear magnetic resonance (1H-NMR) and enzymatic analysis to have the structures:

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Abbreviations

Con A:

Concanavalin A

DEAE:

diethylaminoethyl

Gal:

galactose

Glc:

glucose

HPLC:

high-performance liquid chromatography

M r :

apparent molecular mass

NMR:

nuclear magnetic resonance

pI:

isoelectric point

SDS-PAGE:

sodium dodecyl sulphate-polyacrylamide gel electrophoresis

Xyl:

xylose

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Authors and Affiliations

  1. Department of Biological and Molecular Sciences, FK9 4LA, Stirling, UK

    Cristina Fanutti & J. S. Grant Reid

  2. Unilever Research Laboratories, Colworth House, MK44 1LQ, Sharnbrook, Bedford, UK

    Michael J. Gidley

Authors
  1. Cristina Fanutti
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  2. Michael J. Gidley
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  3. J. S. Grant Reid
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Additional information

Much of the work reported in this paper was carried out with the aid of the European Community's “Science Stimulation Action” (Contract No. ST2P-0250-UK), and we wish to record our appreciation of this support.

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Fanutti, C., Gidley, M.J. & Reid, J.S.G. A xyloglucan-oligosaccharide-specific α-d-xylosidase or exo-oligoxyloglucan-α-xylohydrolase from germinated nasturtium (Tropaeolum majus L.) seeds. Planta 184, 137–147 (1991). https://doi.org/10.1007/BF00208247

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