Characterization of S-adenosyl-L-methionine:(iso)eugenol O-methyltransferase involved in floral scent production in Clarkia breweri

Archives of Biochemistry and Biophysics
J Wang, E Pichersky

Abstract

Eugenol, isoeugenol, methyleugenol, and isomethyleugenol are volatiles found in the floral scent of Clarkia breweri. With their distinct aromas, they are used in many perfumes and food seasonings. Here we report the purification and characterization of (iso)eugenol O-methyltransferase (IEMT), the enzyme that methylates eugenol or isoeugenol to make methyleugenol or isomethyleugenol, respectively, using S-adenosyl-L-methionine as the methyl donor. C. breweri IEMT was copurified with caffeic acid O-methyltransferase (COMT) from petals and purified to homogeneity from a bacterial expression system. IEMT is active as a homodimer with a subunit molecular mass of 40 kDa. It is stable at temperatures up to 35 degrees C. It shows optimum activity at pH 7.5, and it does not require any cofactors for enzymatic activity. Plant-purified IEMT has K(m) values of 7 and 58 microM for eugenol and isoeugenol, respectively, and 27 microM for SAM (30, 74, and 19 microM, respectively, for the plant IEMT expressed in Escherichia coli). By substituting coding regions from COMT into IEMT, it was determined that the regions in IEMT involved in substrate specificity are located in the first half of the protein sequence and that a small segment of 82 ami...Continue Reading

References

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Citations

Jun 17, 1998·The Plant Journal : for Cell and Molecular Biology·N DudarevaE Pichersky
Aug 3, 2004·Plant Physiology·Jennifer BoatrightNatalia Dudareva
Oct 17, 2008·Genome Génome / Conseil National De Recherches Canada·Je Hyeong JungYong Weon Seo
May 3, 2005·Annual Review of Plant Biology·David R Gang
Oct 20, 2005·Journal of Bioscience and Bioengineering·Shuiqin WuYutaka Ebizuka
Mar 5, 2014·Plant, Cell & Environment·Joëlle K MuhlemannNatalia Dudareva
Apr 6, 2017·The Plant Journal : for Cell and Molecular Biology·Yar-Khing YaukRoss G Atkinson
Jul 11, 2006·Plant Molecular Biology·Huaijun Michael LiFaith C Belanger
Nov 3, 2005·The Plant Journal : for Cell and Molecular Biology·Jihong Wang, Vincenzo De Luca
Apr 24, 2015·The Plant Journal : for Cell and Molecular Biology·Yar-Khing YaukRoss G Atkinson
Oct 17, 2014·The Plant Journal : for Cell and Molecular Biology·Kelsey J R P ByersHarvey D Bradshaw
Jan 18, 2018·Bioscience, Biotechnology, and Biochemistry·Takao Koeduka
Oct 31, 2009·The Journal of Biological Chemistry·Mohammad-Wadud Bhuiya, Chang-Jun Liu
May 1, 2021·International Journal of Molecular Sciences·Daniela P HerreraAndreas Schüller
Jul 23, 1999·Archives of Biochemistry and Biophysics·J Wang, E Pichersky
Nov 9, 2000·Biochemical Systematics and Ecology·T J Barkman
Mar 11, 2000·Plant Physiology·N Dudareva, E Pichersky
Dec 23, 2016·Journal of Agricultural and Food Chemistry·Ross G Atkinson

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