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Citral dehydrogenase involved in geraniol oxidation pathway: purification, characterization and kinetic studies from Persicaria minor (Polygonum minus Huds.)

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Abstract

Plants emit semiochemicals as alarm signals upon attack by herbivores or insect pests. Complex insect-plant interaction through alarm pheromones can be manipulated to improve crop protection. Geraniol, citral and geranic acid are monoterpenoid compounds from plants and they play a role as semiochemical alarm pheromones. In plants, the oxidation of geraniol into geranic acid is catalyzed by two oxidoreductases, geraniol dehydrogenase and citral dehydrogenase. In this study, citral dehydrogenase isoenzymes from Persicaria minor (Polygonum minus) leaves were purified to homogeneity and characterized. Enzyme purification through Toyopearl GigaCap Q-650 M column chromatography at pH 7.5 produced two activity peaks, suggesting the existence of two citral dehydrogenase isoenzymes. Both isoenzymes were different in isoelectric point and kinetic parameters but similar in pH and optimal temperature as well as in substrate specificity. Findings from this study will provide a basic understanding for the development of recombinant production of these particular enzymes. Further studies on molecular structure involved could be exploited in transgenic plant as an integrated pest management strategy.

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Abbreviations

pI:

Isoelectric point

IPM:

Integrated pest management

2-ME:

2-Mercaptoethanol

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Acknowledgements

We wish to thank Prof. Dr. Eduardo Mendoza (University of the Philippines Los Banos), Mohd Fauzi Abd Razak (UKM), Nor Ain Shahajar Ahmad Sohdi (UKM), Ahmad Faris Kamaruzzaman (UKM) and Saidi Adha Suhaimi (UKM) for technical assistance and guidance. The acknowledgement goes to the Ministry of Higher Education Malaysia and International Islamic University Malaysia for awarding a scholarship under Skim Latihan Akademik Bumiputera/IPTA (SLAB/SLAI) scheme to NRNAG.

Funding

This project is funded by Ministry of Higher Education Malaysia via the Fundamental Research Grant Scheme FRGS/1/2012/ST04/UKM/02/1 awarded to MH.

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Correspondence to Maizom Hassan.

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Supplementary material 1 (PDF 381 kb)

13562_2017_410_MOESM2_ESM.pdf

S1 Fig Possible pathway of enzymatic production of geranic acid in plant. S2 Fig List of substrates used for determination of substrate specificity. S3 Fig Chromatographic separation of citral dehydrogenase on a GigaCap Q Toyopearl 650 M column. S4 Fig Native-PAGE analysis of purified citral dehydrogenase I (98 µg) and citral dehydrogenase II (86 µg) from P. minor with (A) silver staining and (B) activity staining. S5 Fig pI values of citral dehydrogenase I and citral dehydrogenase II (PDF 327 kb)

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Nik-Abdul-Ghani, NR., Mohamed-Hussein, ZA. & Hassan, M. Citral dehydrogenase involved in geraniol oxidation pathway: purification, characterization and kinetic studies from Persicaria minor (Polygonum minus Huds.). J. Plant Biochem. Biotechnol. 27, 1–12 (2018). https://doi.org/10.1007/s13562-017-0410-1

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  • DOI: https://doi.org/10.1007/s13562-017-0410-1

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