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Expression of mung bean pectin acetyl esterase in potato tubers: effect on acetylation of cell wall polymers and tuber mechanical properties

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An Erratum to this article was published on 07 April 2012

Abstract

A mung bean (Vigna radiata) pectin acetyl esterase (CAA67728) was heterologously expressed in tubers of potato (Solanum tuberosum) under the control of the granule-bound starch synthase promoter or the patatin promoter in order to probe the significance of O-acetylation on cell wall and tissue properties. The recombinant tubers showed no apparent macroscopic phenotype. The enzyme was recovered from transgenic tubers using a high ionic strength buffer and the extract was active against a range of pectic substrates. Partial in vivo de-acetylation of cell wall polysaccharides occurred in the transformants, as shown by a 39% decrease in the degree of acetylation (DA) of tuber cell wall material (CWM). Treatment of CWM using a combination of endo-polygalacturonase and pectin methyl esterase extracted more pectin polymers from the transformed tissue compared to wild type. The largest effect of the pectin acetyl esterase (68% decrease in DA) was seen in the residue from this extraction, suggesting that the enzyme is preferentially active on acetylated pectin that is tightly bound to the cell wall. The effects of acetylation on tuber mechanical properties were investigated by tests of failure under compression and by determination of viscoelastic relaxation spectra. These tests suggested that de-acetylation resulted in a stiffer tuber tissue and a stronger cell wall matrix, as a result of changes to a rapidly relaxing viscoelastic component. These results are discussed in relation to the role of pectin acetylation in primary cell walls and its implications for industrial uses of potato fibres.

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Abbreviations

ANOVA:

Analysis of variance

CWM:

Cell wall material

DA:

Degree of acetylation expressed as mol HOAc/mol UA × 100

endoPG:

Endopolygalacturonase

GBSS:

Granule-bound starch synthase

HG:

Homogalacturonan

HOAc:

Acetic acid

IPTG:

Isopropyl®-d-1-thiogalactopyranoside

PAE:

Pectin acetyl esterase

PAT:

Patatin

PME:

Pectin methyl esterase

pnp-OAc:

p-Nitrophenol acetate

RG-I:

Rhamnogalacturonan-I

UA:

Uronic acid

τ:

Relaxation time

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Acknowledgments

This work was supported by grants from the VKR Research Centre Pro-Active Plants, by the EU-project number 211982 ‘Renewall’, by the Danish National Advanced Technology Foundation, and by the Office of Science, Office of Biological and Environmental Research, of the US Department of Energy under Contract No. DE-AC02-05CH11231. Morten L. Stephensen is acknowledged for skilful technical assistance.

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Author notes

  1. Caroline Orfila

    Present address: School of Food Science and Nutrition, University of Leeds, Leeds, LS2 9JT, UK

  2. Florence Dal Degan

    Present address: ZEALAND Pharma A/S, Smedeland 36, 2600, Glostrup, Denmark

  3. Bodil Jørgensen

    Present address: Department of Agriculture and Ecology, Copenhagen University, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark

Authors and Affiliations

  1. Department of Plant Biology and Biotechnology, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark

    Caroline Orfila, Florence Dal Degan, Bodil Jørgensen, Henrik Vibe Scheller & Peter Ulvskov

  2. Feedstocks Division, Joint BioEnergy Institute, Lawrence Berkeley National Laboratory, Emeryville, CA, 94608, USA

    Henrik Vibe Scheller

  3. Department of Plant and Microbial Biology, University of California, Berkeley, CA, 94720, USA

    Henrik Vibe Scheller

  4. Department of Biological Sciences, Stanford University, Serra Street, Stanford, CA, 94305, USA

    Peter M. Ray

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  1. Caroline Orfila
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Correspondence to Peter Ulvskov.

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Orfila, C., Degan, F.D., Jørgensen, B. et al. Expression of mung bean pectin acetyl esterase in potato tubers: effect on acetylation of cell wall polymers and tuber mechanical properties. Planta 236, 185–196 (2012). https://doi.org/10.1007/s00425-012-1596-z

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