Planta

, Volume 224, Issue 5, pp 1091–1102 | Cite as

LeMAN4 endo-β-mannanase from ripe tomato fruit can act as a mannan transglycosylase or hydrolase

  • Roswitha Schröder
  • Teresa F. Wegrzyn
  • Neelam N. Sharma
  • Ross G. Atkinson
Original Article

Abstract

Mannan transglycosylases are cell wall enzymes able to transfer part of the mannan polysaccharide backbone to mannan-derived oligosaccharides (Schröder et al. in Planta 219:590–600, 2004). Mannan transglycosylase activity was purified to near homogeneity from ripe tomato fruit. N-terminal sequencing showed that the dominant band seen on SDS-PAGE was identical to LeMAN4a, a hydrolytic endo-β-mannanase found in ripe tomato fruit (Bewley et al. in J Exp Bot 51:529–538, 2000). Recombinant LeMAN4a protein expressed in Escherichia coli exhibited both mannan hydrolase and mannan transglycosylase activity. Western analysis of ripe tomato fruit tissue using an antibody raised against tomato seed endo-β-mannanase revealed four isoforms present after 2D-gel electrophoresis in the pH range 6–11. On separation by preparative liquid isoelectric focussing, these native isoforms exhibited different preferences for transglycosylation and hydrolysis. These results demonstrate that endo-β-mannanase has two activities: it can either hydrolyse mannan polysaccharides, or in the presence of mannan-derived oligosaccharides, carry out a transglycosylation reaction. We therefore propose that endo-β-mannanase should be renamed mannan transglycosylase/hydrolase, in accordance with the nomenclature established for xyloglucan endotransglucosylase/hydrolase. The role of endo-acting mannanases in modifying the structure of plant cell walls during cell expansion, seed germination and fruit ripening may need to be reinterpreted in light of their potential action as transglycosylating or hydrolysing enzymes.

Keywords

Cell wall Endo-β-mannanase Hydrolase Mannan Tomato Transglycosylase 

Abbreviations

GGM

Galactoglucomannan

GGMOs

Galactoglucomannan-derived oligosaccharides

G2M5

Di-galactosyl mannopentaose

M1–M5

Mannose to mannopentaose oligosaccharides

MTH

Mannan endotransglycosylase/hydrolase

XTH

Xyloglucan endotransglucosylase/hydrolase

Notes

Acknowledgments

We thank Dr. Y. Morohashi, Tokyo University of Agriculture and Technology, Japan, for kindly providing the tomato endo-β-mannanase antibody. We also thank Status Produce, Pukekohe, New Zealand for generously supplying us with tomatoes; Di Barraclough for her invaluable advice and support during 2D-gel electrophoresis; the protein sequencing team at Ruakura and Dave Greenwood for their help with interpretation of the mass spectrometry data; and Sandie Fry for technical assistance. Our gratitude goes to Matt Templeton, Andrew Kralicek and Dave Greenwood for critically reviewing the manuscript. This project was funded by the Foundation for Research, Science and Technology (C06X0202), New Zealand.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Roswitha Schröder
    • 1
  • Teresa F. Wegrzyn
    • 1
  • Neelam N. Sharma
    • 1
  • Ross G. Atkinson
    • 1
  1. 1.Mt. Albert Research CentreThe Horticulture and Food Research Institute of New ZealandAucklandNew Zealand

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