PLANT PHYSIOLOGY , Vol 110, Issue 3 791-799, Copyright © 1996 by American Society of Plant Biologists
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BIOCHEMISTRY AND ENZYMOLOGY |
Aromatic Polyketide Synthases (Purification, Characterization, and Antibody Development to Benzalacetone Synthase from Raspberry Fruits)
W. Borejsza-Wysocki and G. Hrazdina
Institute of Food Science, Cornell University, Geneva, New York 14456-0462
p-Hydroxyphenylbutan-2-one, the characteristic aroma compound of
raspberries (Rubus idaeus L.), is synthesized from p-coumaryl-coenzyme A
and malonyl-coenzyme A in a two-step reaction sequence that is catalyzed by
benzalacetone synthase and benzalacetone reductase (W. Borejsza-Wysocki and
G. Hrazdina [1994] Phytochemistry 35: 623-628). Benzalacetone synthase
condenses one malonate with p-coumarate to form the pathway intermediate
p-hydroxyphenylbut-3-ene-2-one (p-hydroxybenzalacetone) in a reaction that
is similar to those catalyzed by chalcone and stilbene synthases. We have
obtained an enzyme preparation from ripe raspberries that was
preferentially enriched in benzalacetone synthase (approximately 170-fold)
over chalcone synthase (approximately 14-fold) activity. This preparation
was used to characterize benzalacetone synthase and to develop polyclonal
antibodies in rabbits. Benzalacetone synthase showed similarity in its
molecular properties to chalcone synthase but differed distinctly in its
substrate specificity, response to 2-mercaptoethanol and ethylene glycol,
and induction in cell-suspension cultures. The product of the enzyme,
p-hydroxybenzalacetone, inhibited mycelial growth of the raspberry pathogen
Phytophthora fragariae var rubi at 250 [mu]M. We do not know whether the
dual activity in the benzalacetone synthase preparation is the result of a
bifunctional enzyme or is caused by contamination with chalcone synthase
that was also present. The rapid induction of the enzyme in cell-suspension
cultures upon addition of yeast extract and the toxicity of its product,
p-hydroxybenzalacetone, to phytopathogenic fungi also suggest that the
pathway may be part of a plant defense response.