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Substrate specificity of the violaxanthin de-epoxidase of the primitive green alga Mantoniella squamata (Prasinophyceae)

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Abstract

The substrate specificity of the enzyme violaxanthin de-epoxidase (VDE) of the primitive green alga Mantoniella squamata (Prasinophyceae) was tested in in vitro enzyme assays employing the following xanthophyll mono-epoxides: antheraxanthin (Ax), diadinoxanthin (Ddx), lutein-epoxide (LE), cryptoxanthin-epoxide (CxE), 9-cis neoxanthin (cNx), all-trans neoxanthin (Nx), and xanthophyll di-epoxides: 9-cis violaxanthin (cVx), all-trans violaxanthin (Vx), cryptoxanthin-di-epoxide (CxDE). The data presented in this study show that the VDE of M. squamata not only exhibits a low affinity for the mono-epoxide Ax, as has been reported by R. Frommolt et al. (2001, Planta 213:446–456), but has a reduced substrate affinity for the mono-epoxides Ddx, LE, CxE, and Nx as well. On the other hand, xanthophylls with a second epoxy-group (Vx, CxDE) can be de-epoxidized with a higher efficiency. Such a preference for xanthophyll di-epoxides cannot be observed for the higher-plant VDE, where, in general, no marked differences in the pigment de-epoxidation rates between xanthophyll mono- and di-epoxides are visible. Despite this substantial difference between the VDEs of M. squamata and S. oleracea there are also features common to both enzymes. Neither VDE is able to convert xanthophylls with a 9-cis configuration in the acyclic polyene chain and both rely on substrates in the all-trans configuration. Both enzymes furthermore exhibit a dependence of enzyme activity on the polarity of the substrate. Highly polar (Nx) or non-polar (CxE) xanthophylls are de-epoxidized with greatly reduced rates in comparison to substrates with an intermediate polarity (Vx, Ax, LE, Ddx). This dependence on substrate polarity becomes more obvious when the higher-plant VDE is examined, as the substrate affinity of the VDE of M. squamata is more strongly influenced by the existence or absence of a second epoxy-group. In summary, the data presented in this study underline the fact that different VDEs, although in general catalyzing the same reaction sequence, are functionally diverse.

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Abbreviations

Ax:

antheraxanthin

Cx:

cryptoxanthin

CxDE:

cryptoxanthin-5,6,5′,6′-di-epoxide

CxE:

cryptoxanthin-5,6-epoxide

Ddx:

diadinoxanthin

Dtx:

diatoxanthin

L:

lutein

LE:

lutein-5,6-epoxide

MGDG:

monogalactosyldiacylglycerol

Nx:

all-trans neoxanthin

cNx:

9-cis neoxanthin

dNx:

de-epoxy Nx

VDE:

violaxanthin de-epoxidase

Vx:

all-trans violaxanthin

cVx:

9-cis violaxanthin (violeoxanthin)

Zx:

zeaxanthin

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Acknowledgements

Thanks are due to Prof. C. Wilhelm and Dr. T. Jakob, Institut für Botanik, Universität Leipzig, Leipzig, Germany for critical reading of the manuscript, and to Dr. M. Lohr, Institut für Allgemeine Botanik, Johannes Gutenberg-Universität, Mainz, Germany for his help optimizing the de-epoxidation assay.

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  1. Institut für Botanik–Pflanzenphysiologie, Universität Leipzig, Johannisallee 21–23, 04103, Leipzig, Germany

    Reimund Goss

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Goss, R. Substrate specificity of the violaxanthin de-epoxidase of the primitive green alga Mantoniella squamata (Prasinophyceae). Planta 217, 801–812 (2003). https://doi.org/10.1007/s00425-003-1044-1

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