When grown on Czapek-Dox agar, Penicillium brevicompactum produced mycophenolic acid after a vegetative mycelium had been formed and as aerial hyphae were developing. Nutrients were still plenteous in the agar when the synthesis began. If aerial hyphal development was prevented by placing a dialysis membrane over the growing fungus, no mycophenolic acid was produced. When the dialysis membrane was peeled back and, as a consequence, production of aerial hyphae began, mycophenolic acid biosynthesis was observed. We concluded that mycophenolic acid was produced only by P. brevicompactum colonies that possessed an aerial mycelium.
Penicillium brevicompactum produces mycophenolic acid as it grows vegetatively, not only on a simple medium where growth is slow but also on a richer medium where growth is less restricted. The implications of this finding on the association of fungal secondary metabolism with the idiophase in liquid and solid culture are discussed.
Formation of 7 alpha-hydroxycholesterol by rat liver microsomes was quantitated using a gas chromatograph-mass spectrometer (GC/MS) operated in selected ion monitoring (SIM) mode. Microsomes from normal rat livers incubated for different periods were found to yield increased 7 alpha-hydroxycholesterol with time. This was also true when incubations contained Tween-80, but in this instance, the rate of 7 alpha-hydroxycholesterol production was lower and dependent on the concentration of Tween used. Similarly, Triton X-100, Renex-30, Kyro EOB, Cutscum, and Emulgen 911 all lowered the formation of 7 alpha-hydroxycholesterol by rat liver microsomes, whereas Triton WR-1339 stimulated its production. Analysis of data obtained from following the enzyme reaction over an extended period using an integrated Michaelis-Menten equation indicated the enzyme possesses a very significant affinity for the product (Ks greater than Kp). Similar analysis shows that Tween-80 is a noncompetitive inhibitor of the enzyme.
The naphthalenones asparvenone and its 6-O-methyl ether are produced by Aspergillus parvulus during vegetative growth in batch-mode, shaken cultures in a glucose-glycine-salts medium. The secondary metabolites first appear around the time when hyphae of developing spores begin to make contact with each other, i.e. at a very early stage of culture development. Resuspension in new medium of mycelium that is producing the naphthalenones fails to inhibit production. When A. parvulus is grown in parallel on a glucose-glycine-salts medium and on a glucose-malt-peptone medium, the naphthalenones appear first in the richer medium despite the fact that biomass proliferation is relatively unrestricted in that medium. Radiolabelled [1-14C]-acetate is taken up quickly by the fungus (k = 0.09 min-1) and is incorporated effectively into the naphthalenones.
When Penicillium patulum was grown on Czapek-Dox agar, 6-methylsalicylic acid was produced as an aerial mycelium was forming. Nutrients were often plentiful in the medium when biosynthesis began. If the formation of an aerial mycelium was prevented by growing the fungus between two sheets of dialysis membrane, no 6-methylsalicylic acid was produced even when nutrients were completely consumed. If the upper sheet of dialysis membrane was stripped off cultures of the latter type, an aerial mycelium formed; concomitantly, 6-methylsalicylic acid biosynthesis was observed. We conclude that 6-methylsalicylic acid was produced only by P. patulum colonies that possessed an aerial mycelium.
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