In our screening for bioactive secondary metabolites from microorganisms, 1 fungal contamination of the S. argillaceus M7OII 2 was found to produce an intensive non-polar yellow pigment. Working up of the strain resulted in the isolation of all-trans (E)-8-methyl-13-phenyltrideca-4,6,8,10,12-pentaen-3-one, also called asperenone, (1). A literature search revealed that (1) has been isolated from both Aspergillus awamori and niger. 3,4 It exhibited an inhibitor of soybean 15-lipoxygenase and human platelet aggregation.5 Spectroscopic studies of (1) have indicated that the pigment is a substituted phenyltridecapentaene-3-one, with methyl group located somewhere on the side-chain. In 1996, the location of the methyl group was suggested using NMR spectroscopy.
6In addition, the stereochemistry was assumed to be all-trans (E) based on the relative extinction coefficients of bands associated with the vibrational fine structure of the absorbing molecule in its visible and UV spectra. In order to obtain detailed information on the structural conformation of 1, and to confirm the geometry, an X-ray structure determination was carried out.The fungal contamination of the S. argillaceus M7OII was isolated and purified on agar plates with the M2 medium (glucose 4 g/l, malt extract 10 g/l and yeast extract 4 g/l) at 28˚C for 48 -72 h, where it grew with a thick greenish aerial mycelium and yellow agar coloration.Thirty 250 cm 3 Erlenmeyer flasks, each containing 100 mL of M2 medium, were inoculated with a well-grown agar subculture and incubated with 110 rpm at 28˚C for 3 days. The yellow culture broth was centrifuged at 4200 rpm for 20 min. The resulting broth and mycelial cake were extracted three times with ethyl acetate. Since the extracts exhibited a similar composition by TLC, they were combined and evaporated to dryness under a vacuum at 30˚C. The crude residue (ca. 0.9 g) was pre-separated into two fractions on Sephadex LH-20 (4 ¥ 120 cm, CH2Cl2/50% MeOH). The first fraction (145 mg) containing the yellow pigment 1 was finally purified by preparative HPLC (column, mBondapak C18 radial compression cartridge, PrepPak cartridge, 19 K 150 mm, Waters; eluent water and acetonitrile (gradient from 35 to 100% in 30 min); flow rate, 7 mL/min -1 ) to give 6.9 mg of 1.
X-ray