Plants, such as Arabidopsis thaliana and Selaginella lepidophylla, contain genes homologous with the trehalose-6-phosphate synthase (TPS) genes of bacteria and fungi. Most plants do not accumulate trehalose with the desert resurrection plant S. lepidophylla, being a notable exception. Overexpression of the plant genes in a Saccharomyces cerevisiae tps1 mutant results in very low TPS-catalytic activity and trehalose accumulation. We show that truncation of the plant-specific N-terminal extension in the A. thalianaAtTPS1 and S. lepidophyllaSlTPS1 homologues results in 10–40-fold higher TPS activity and 20–40-fold higher trehalose accumulation on expression in yeast. These results show that the plant TPS enzymes possess a high-potential catalytic activity. The growth defect of the tps1 strain on glucose was restored, however, the proper homoeostasis of glycolytic flux was not restored, indicating that the plant enzymes were unable to substitute for the yeast enzyme in the regulation of hexokinase activity. Further analysis of the N-terminus led to the identification of two conserved residues, which after mutagenesis result in strongly enhanced trehalose accumulation upon expression in yeast. The plant-specific N-terminal region may act as an inhibitory domain allowing modulation of TPS activity.
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August 2002
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Research Article|
August 15 2002
Truncation of Arabidopsis thaliana and Selaginella lepidophylla trehalose-6-phosphate synthase unlocks high catalytic activity and supports high trehalose levels on expression in yeast
Patrick Van DIJCK;
Patrick Van DIJCK
1
∗Laboratorium voor Moleculaire Celbiologie, Katholieke Universiteit Leuven and Vlaams Interuniversitair Instituut voor Biotechnologie (VIB), Kasteelpark Arenberg 31, B-3001 Leuven-Heverlee, Flanders, Belgium
4To whom correspondence should be addressed (e-mail johan.thevelein@bio.kuleuven.ac.be).
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José O. MASCORRO-GALLARDO;
†Departamento de Biología Molecular de Plantas, Instituto de Biotecnología-UNAM, Av. Universidad 2001, Col. Chamilpa, Cuernavaca, Morelos 62210, Mexico
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Martien De BUS;
Martien De BUS
∗Laboratorium voor Moleculaire Celbiologie, Katholieke Universiteit Leuven and Vlaams Interuniversitair Instituut voor Biotechnologie (VIB), Kasteelpark Arenberg 31, B-3001 Leuven-Heverlee, Flanders, Belgium
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Katrien ROYACKERS;
Katrien ROYACKERS
∗Laboratorium voor Moleculaire Celbiologie, Katholieke Universiteit Leuven and Vlaams Interuniversitair Instituut voor Biotechnologie (VIB), Kasteelpark Arenberg 31, B-3001 Leuven-Heverlee, Flanders, Belgium
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Gabriel ITURRIAGA;
Gabriel ITURRIAGA
3
∗Laboratorium voor Moleculaire Celbiologie, Katholieke Universiteit Leuven and Vlaams Interuniversitair Instituut voor Biotechnologie (VIB), Kasteelpark Arenberg 31, B-3001 Leuven-Heverlee, Flanders, Belgium
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Johan M. THEVELEIN
Johan M. THEVELEIN
4
∗Laboratorium voor Moleculaire Celbiologie, Katholieke Universiteit Leuven and Vlaams Interuniversitair Instituut voor Biotechnologie (VIB), Kasteelpark Arenberg 31, B-3001 Leuven-Heverlee, Flanders, Belgium
4To whom correspondence should be addressed (e-mail johan.thevelein@bio.kuleuven.ac.be).
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Publisher: Portland Press Ltd
Received:
April 03 2002
Revision Received:
April 23 2002
Accepted:
April 29 2002
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2002
2002
Biochem J (2002) 366 (1): 63–71.
Article history
Received:
April 03 2002
Revision Received:
April 23 2002
Accepted:
April 29 2002
Citation
Patrick Van DIJCK, José O. MASCORRO-GALLARDO, Martien De BUS, Katrien ROYACKERS, Gabriel ITURRIAGA, Johan M. THEVELEIN; Truncation of Arabidopsis thaliana and Selaginella lepidophylla trehalose-6-phosphate synthase unlocks high catalytic activity and supports high trehalose levels on expression in yeast. Biochem J 15 August 2002; 366 (1): 63–71. doi: https://doi.org/10.1042/bj20020517
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