title: Dual positional and stereospecificity of lipoxygenase isoenzymes from germinating barley (green malt): biotransformation of free and esterified linoleic acid.
abstract: The lipoxygenase isoenzymes LOX1 and LOX2 from green malt were separated by isoelectric focusing, and their catalytic properties regarding complex lipids as substrates were characterized. The regio- and stereoisomers of hydroperoxy octadecadienoates (HPODE) resulting from LOX1 and LOX2 enzymatic transformations of linoleic acid, methyl linoleate, linoleic acid glycerol esters monolinolein, dilinolein, and trilinolein, and 1-palmitoyl-2-linoleoyl-glycero-3-phosphocholine (PamLinGroPCho) were determined. In addition, biotransformations of polar and nonpolar lipids extracted from malt were performed with LOX1 and LOX2. The results show that LOX2 catalyzes the oxidation of esterified fatty acids at a higher rate and is more regioselective than LOX1. The dual position specificity of LOX2 (9-HPODE:13-HPODE) with trilinolein as the substrate (6:94) was higher than the resultant ratio (13:87) when free linoleic acid was transformed. A high (S)-enantiomeric excess of 13-HPODE was analyzed with all esterified substrates confirming the formation of 13-HPODE through the LOX2 enzyme; however, 9-HPODE detected after LOX2 biotransformations showed (R)-enantiomeric excesses. PamLinGroPCho was oxygenated by LOX1 with the highest regio- and stereoselectivities among the applied substrates.
authors.affiliation: Institut für Biotechnologie, Molekularanalytik, Technische Universität Berlin, Seestrasse 13, D-13353 Berlin, Germany. Leif-A.Garbe@TU-Berlin.de
authors.forename: Leif-Alexander
authors.initials: LA
authors.lastname: Garbe
authors.forename: Roberto
authors.initials: R
authors.lastname: Barbosa de Almeida
authors.forename: Renate
authors.initials: R
authors.lastname: Nagel
authors.forename: Karl
authors.initials: K
authors.lastname: Wackerbauer
authors.forename: Roland
authors.initials: R
authors.lastname: Tressl
mesh_terms: D004951:Esterification; D018525:Germination; D001467:Hordeum / Q000201:enzymology*; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D019787:Linoleic Acid / Q000378:metabolism*; D008041:Linoleic Acids / Q000378:metabolism; D008054:Lipid Peroxides / Q000378:metabolism; D008084:Lipoxygenase / Q000302:isolation & purification / Q000378:metabolism*; D012639:Seeds / Q000201:enzymology* / Q000254:growth & development; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D007527:Isoenzymes; D008041:Linoleic Acids; D008054:Lipid Peroxides; C038649:13-hydroperoxy-9,11-octadecadienoic acid; C049606:9-hydroperoxy-11,12-octadecadienoic acid; D019787:Linoleic Acid; D008084:Lipoxygenase