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Purification and biochemical characterisation of a glucose-6-phosphate dehydrogenase from the psychrophilic green alga Koliella antarctica

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Abstract

Psychrophilic organisms have evolved a number of modifications of cellular structures to survive in the cold environment; among them it is worth noting an increased efficiency of enzymes at lower temperatures. Glucose-6-phosphate dehydrogenase (G6PDH; EC 1.1.1.49) was purified and characterised from the psychrophilic green alga Koliella antarctica (Trebouxiophyceae, Chlorophyta) from the Ross Sea (Antarctica). It was possible to isolate a single G6PDH using biochemical strategies; its maximum activity was measured at 35 °C, and the enzyme showed an E a of 39.6 kJ mol−1. This protein reacted with antibodies raised against higher plants plastidic isoforms. KaG6PDH showed peculiar kinetic properties, with a K iNADPH value lower than \( K_{{{\text{mNADP}}^{ + } }} \). Notably, catalytic activity was inactivated in vitro by DTT and chloroplastic thioredoxin f. These biochemical properties of G6PDH are discussed with respect to higher plant G6PDHs and the adaptation of K. antarctica to polar low-temperature environment.

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Abbreviations

DTT:

Dithiothreitol

G6P:

Glucose-6-phosphate

G6PDH:

Glucose-6-phosphate dehydrogenase

Ka :

Koliella antarctica

NaTT:

Sodium tetrathionate

OPPP:

Oxidative pentose phosphate pathway

PAGE:

Polyacrylamide gel electrophoresis

SDS:

Sodium dodecyl sulfate

Tris:

Tris-(hydroxymethyl)-aminomethane

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Acknowledgments

We are grateful to Prof. Andreoli, University of Padova (Italy), for the gift of Koliella antarctica. Thanks to Dr. M. Caiazzo and Dr. O. Lobosco for the batch growth of K. antarctica at the Plant Biology Section of the Dept of Biological Sciences at the University of Naples. We thank very much Prof. A. von Schaewen (Munster, Germany) for the generous gift of G6PDH antibodies. Dr. K. Chibani (University of Lorraine, Nancy, France) for the purified recombinant spinach thioredoxin f. We are in debt with Dr. M. Chiurazzi (IGB-CNR “Buzzati Traverso”, Naples) for the help and assistance to obtain the partial sequence of the G6PDH active site of K. antarctica. Many thanks to J.-P. Jacquot (University of Lorraine, Nancy, France), for reading the manuscript, the helpful discussion, and suggestions.

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Authors and Affiliations

  1. Dipart. Biologia Strutturale e Funzionale, Università di Napoli “Federico II”, Complesso Universitario di Monte Sant’Angelo, Via Cinthia 6, 80126, Naples, Italy

    Myriam Ferrara, Manuela Cardi & Sergio Esposito

  2. Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Science Vienna, University and Research Center UFT-Campus Tulln, Konrad Lorenz Strasse 24, A-3430, Tulln/Donau, Austria

    Gea Guerriero

Authors
  1. Myriam Ferrara
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  2. Gea Guerriero
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  3. Manuela Cardi
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  4. Sergio Esposito
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Corresponding author

Correspondence to Sergio Esposito.

Additional information

Communicated by H. Atomi.

The authors wish to dedicate this paper to Prof. Carmelo Rigano for his over 40 years of teaching and researching in Plant Physiology at the University of Naples “Federico II”.

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Ferrara, M., Guerriero, G., Cardi, M. et al. Purification and biochemical characterisation of a glucose-6-phosphate dehydrogenase from the psychrophilic green alga Koliella antarctica . Extremophiles 17, 53–62 (2013). https://doi.org/10.1007/s00792-012-0492-6

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