Purification and characterization of a novel NAD(P)+-farnesol dehydrogenase from polygonum minus leaves

Nor Ain Shahajar Ahmad-Sohdi, Ahmad Faris Seman-Kamarulzaman, Zeti Azura Mohamed Hussein, Maizom Hassan

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Juvenile hormones have attracted attention as safe and selective targets for the design and development of environmentally friendly and biorational insecticides. In the juvenile hormone III biosynthetic pathway, the enzyme farnesol dehydrogenase catalyzes the oxidation of farnesol to farnesal. In this study, farnesol dehydrogenase was extracted from Polygonum minus leaves and purified 204-fold to apparent homogeneity by ion-exchange chromatography using DEAE-Toyopearl, SP-Toyopearl, and Super-Q Toyopearl, followed by three successive purifications by gel filtration chromatography on a TSK-gel GS3000SW. The enzyme is a heterodimer comprised of subunits with molecular masses of 65 kDa and 70 kDa. The optimum temperature and pH were 35°C and pH 9.5, respectively. Activity was inhibited by sulfhydryl reagents, metal-chelating agents and heavy metal ions. The enzyme utilized both NAD+ and NADP+ as coenzymes with Km values of 0.74 mM and 40 mM, respectively. Trans, trans-farnesol was the preferred substrate for the P. minus farnesol dehydrogenase. Geometrical isomers of trans, trans-farnesol, cis, trans-farnesol and cis, cis-farnesol were also oxidized by the enzyme with lower activity. The Km values for trans, trans-farnesol, cis, trans-farnesol and cis, cis-farnesol appeared to be 0.17 mM, 0.33 mM and 0.42 mM, respectively. The amino acid sequences of 4 tryptic peptides of the enzyme were analyzed by MALDI-TOF/TOF-MS spectrometry, and showed no significant similarity to those of previously reported farnesol dehydrogenases. These results suggest that the purified enzyme is a novel NAD(P)+-dependent farnesol dehydrogenase. The purification and characterization established in the current study will serve as a basis to provide new information for recombinant production of the enzyme. Therefore, recombinant farnesol dehydrogenase may provide a useful molecular tool in manipulating juvenile hormone biosynthesis to generate transgenic plants for pest control.

Original languageEnglish
Article numbere0143310
JournalPLoS One
Volume10
Issue number11
DOIs
Publication statusPublished - 1 Nov 2015

Fingerprint

Polygonum
Farnesol
farnesol
NAD (coenzyme)
NAD
Purification
Enzymes
leaves
Juvenile Hormones
enzymes
Chromatography
juvenile hormones
Gels
Pest control
Heavy Ions
Sulfhydryl Reagents
Pest Control
farnesol dehydrogenase
Persicaria minor
Genetically Modified Plants

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Purification and characterization of a novel NAD(P)+-farnesol dehydrogenase from polygonum minus leaves. / Ahmad-Sohdi, Nor Ain Shahajar; Seman-Kamarulzaman, Ahmad Faris; Mohamed Hussein, Zeti Azura; Hassan, Maizom.

In: PLoS One, Vol. 10, No. 11, e0143310, 01.11.2015.

Research output: Contribution to journalArticle

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