title: Preparative enzymatic solid phase synthesis of cis(+)-12-oxo-phytodienoic acid - physical interaction of AOS and AOC is not necessary.
abstract: The pathway of jasmonic acid (JA) biosynthesis was established in the 1980s by Vick and Zimmerman but, until now, the preparative biosynthesis of the jasmonic acid precursors 12-oxo-phytodienoic acid (OPDA) and 3-oxo-2-[2'-pentenyl]-cyclopentan-1-octanoic acid (OPC-8:0) in their endogenous and biologically relevant cis(+)-configuration was only possible in small amounts and had to put up with high costs. This was mainly due to the lack of high amounts of pure and enzymatically active allene oxide cyclase (AOC), which is a key enzyme in the biosynthesis of jasmonates in that it releases, in a coupled reaction with allene oxide synthase (AOS), the first cyclic and biological active metabolite - OPDA. We describe here the expression and purification of AOS and AOC and their subsequent coupling to solid matrices to produce an enantioselective, reusable bioreactor for octadecanoid production. With the method described here it is possible to produce optically pure enantiomers of octadecanoids in high amounts in a cost- and time-efficient manner. Furthermore, it could be demonstrated that a physical interaction of AOS and AOC, hitherto postulated to be required for substrate channeling from AOS to AOC, is not necessary for the in vitro cyclization of the unstable epoxide generated by the AOS reaction.
authors.affiliation: Lehrstuhl für Pflanzenphysiologie, Ruhr-Universität Bochum, Universitätsstr. 150, D-44780 Bochum, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Zerbe
authors.forename: Elmar W
authors.initials: EW
authors.lastname: Weiler
authors.forename: Florian
authors.initials: F
authors.lastname: Schaller
mesh_terms: D003517:Cyclopentanes / Q000378:metabolism; D004800:Enzymes, Immobilized; D005231:Fatty Acids, Unsaturated / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D019746:Intramolecular Oxidoreductases / Q000378:metabolism*; D054883:Oxylipins; D011485:Protein Binding; D013237:Stereoisomerism
chemical_list: D003517:Cyclopentanes; D004800:Enzymes, Immobilized; D005231:Fatty Acids, Unsaturated; D054883:Oxylipins; C025999:12-oxophytodienoic acid; C011006:jasmonic acid; D019746:Intramolecular Oxidoreductases; C021377:hydroperoxide isomerase