Conversion of linoleic acid into novel oxylipins by the mushroom Agaricus bisporus.
Oxylipins are associated with important processes of the fungal life cycle, such as spore formation. Here, we report the formation of FA metabolites in Agaricus bisporus. Incubation of a crude extract of lamellae with linoleic acid (18:2) led to the extensive formation of two oxylipins. They were identified as 8(R)-hydroxy-9Z,12Z-octadecadienoic acid (8-HOD) and 8(R),11 (S)-dihydroxy-9Z,12Z-octadecadienoic acid (8,11-diHOD) by using RP-HPLC, GC-MS, IR, GC-MS analysis of diastereomeric derivatives, and 1H NMR and 13C NMR spectroscopy. Neither compound has been reported before in A. bisporus. Oleic (18:1), alpha-linolenic (18:3n-3), and gamma-linolenic (18:3n-6) acids were converted into their 8-hydroxy derivatives as well, and 18:3n-3 was further metabolized to its 8,11-diol derivative. Reactions with [U-13C]18:2 demonstrated that the compounds 8-HOD and 8,11-diHOD were formed from exogenously supplied 18:2. When [U-13C]8-HOD was supplied, it was not converted into 8,11-diHOD, indicating that it was not an intermediate in the formation of 8,11-diHOD. When a crude extract of A. bisporus was incubated under an atmosphere of 16O2/18O2, the two hydroxyl groups of 8,11-diHOD contained either two 180 atoms or two 60 atoms. Species that contained one of each isotope could not be detected. We propose that the formation of the 8,11-dihydroxy compounds occurs through either an 8,11-endoperoxy, an 8-peroxo free radical, or an 8-hydroperoxy intermediate. In the latter case, the reaction should be catalyzed by dioxygenase with novel specificity.
Wadman
Mayken W
MW
Department of Bio-organic Chemistry, Utrecht University, 3584-CH, Utrecht, The Netherlands.
van Zadelhoff
Guus
G
Hamberg
Mats
M
Visser
Tom
T
Veldink
Gerrit A
GA
Vliegenthart
Johannes F G
JF
D000364:Agaricus / Q000201:enzymology / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D005231:Fatty Acids, Unsaturated / Q000096:biosynthesis* / Q000737:chemistry; D008401:Gas Chromatography-Mass Spectrometry; D019787:Linoleic Acid / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D013055:Spectrophotometry, Infrared
D005231:Fatty Acids, Unsaturated; C027369:octadecadienoic acid; D019787:Linoleic Acid