title: Glucosinolate biosynthesis: demonstration and characterization of the condensing enzyme of the chain elongation cycle in Eruca sativa.
abstract: Glucosinolates are a group of sulfur-rich thioglucoside natural products common in the Brassicaceae and related plant families. The first phase in the formation of many glucosinolates involves the chain extension of the amino acid methionine. Additional methylene groups are inserted into the side chain of methionine by a three-step elongation cycle involving 2-oxo acid intermediates. This investigation demonstrated the first step of this chain elongation cycle in a partially-purified preparation from arugula (Eruca sativa). The 2-oxo acid derived from methionine, 4-methylthio-2-oxobutanoic acid, was shown to condense with acetyl-CoA to form 2-(2'-methylthioethyl)malate. The catalyst, designated as a 2-(omega-methylthioalkyl)malate synthase, belongs to a family of enzymes that mediate the condensation of acyl-CoAs with 2-oxo acids, including citrate synthase of the citric acid cycle, and 2-isopropylmalate synthase of leucine biosynthesis. The 2-(omega-methylthioalkyl)malate synthase studied here shares properties with other enzymes of this class, but appears chromatographically distinct and is found only in extracts of plant species producing glucosinolates from chain-elongated methionine derivatives. Although the principal glucosinolates of arugula are formed from methionine that has undergone two rounds of chain elongation to form dihomomethionine, studies with substrates and substrate analogs of different chain lengths showed that the isolated enzyme is responsible only for the condensation step of the first round of elongation.
authors.affiliation: Max Planck Institute for Chemical Ecology, Beutenberg Campus, Hans-Knöll-Strasse 8, D-07745 Jena, Germany.
authors.forename: Kimberly L
authors.initials: KL
authors.lastname: Falk
authors.forename: Christine
authors.initials: C
authors.lastname: Vogel
authors.forename: Susanne
authors.initials: S
authors.lastname: Textor
authors.forename: Stefan
authors.initials: S
authors.lastname: Bartram
authors.forename: Alastair
authors.initials: A
authors.lastname: Hick
authors.forename: John A
authors.initials: JA
authors.lastname: Pickett
authors.forename: Jonathan
authors.initials: J
authors.lastname: Gershenzon
mesh_terms: D015079:2-Isopropylmalate Synthase / Q000378:metabolism; D000105:Acetyl Coenzyme A / Q000737:chemistry / Q000378:metabolism; D019607:Brassicaceae / Q000201:enzymology* / Q000378:metabolism; D002413:Cations, Divalent / Q000737:chemistry / Q000494:pharmacology; D002736:Chloroplasts / Q000201:enzymology; D005961:Glucosinolates / Q000096:biosynthesis* / Q000737:chemistry; D006868:Hydrolysis; D008292:Malate Synthase / Q000737:chemistry / Q000378:metabolism*; D008316:Malonyl Coenzyme A / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods; D008715:Methionine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D013045:Species Specificity; D013379:Substrate Specificity; D014316:Tritium
chemical_list: D002413:Cations, Divalent; D005961:Glucosinolates; D014316:Tritium; D008316:Malonyl Coenzyme A; C014632:2-keto-4-methylthiobutyric acid; D000105:Acetyl Coenzyme A; D008715:Methionine; D015079:2-Isopropylmalate Synthase; D008292:Malate Synthase