0000000000188440
AUTHOR
Y. Hathout
High-performance liquid chromatographic study of the regulation of phospholipid metabolism in cultured adrenocortical cells
Abstract A rapid high-performance liquid chromatographic (HPLC) method for the separation of phospholipids was developed for minute samples of total lipids (ca. 200 μg). The method was applied to the study of the phospholipid metabolism in adrenocortical cell cultures. A complete separation of the different cellular phospholipid classes was achieved in 40 min. Good resolution of the phospholipid peaks was obtained, which allowed the collection of each individual class of phospholipids for further analysis of radioactivity and fatty acid composition by gas chromatography. When cells were incubated with [U-14C]glycerol or [U-14C]palmitate the bulk of the radioactivity was found in cellular ph…
Production of 6-pentyl-?-pyrone byTrichoderma harzianum from 18?n fatty acid methyl esters
Biosynthesis of 6-pentyl-α-pyrone byTrichoderma harzianum in two different media was evaluated. Best yields were found in nitrogen deficient medium (C/N=60). Limited growth seems to favour the production of this lactone. When fungal cells, precultured in low nitrogen medium, were incubated on methyl ricinoleate (10 g/l, C/N=60) an increase in 6-pentyl-α-pyrone production was observed in comparison with the media containing methyl oleate or methyl linoleate.
Quantitative separation of Trichoderma lipid classes on a bonded phase column.
Bond Elut aminopropyl columns were used to purify the different lipid classes of Trichoderma harzianum and Trichoderma viride. This methodology permitted good separation of the fungal lipid classes in less time than traditional techniques. The incorporation of [1 (14)C]linoleic acid into neutral lipids, free fatty acids and phospholipids was quantified for both strains. The fatty acid profile of the different lipid classes of these fungal strains was determined as a function of culture time.
Aldosterone biosynthesis induced by ACTH and angiotensin II in newborn rat adrenocortical cells transfected by c-EJ-Ha-ras oncogene
Abstract Adrenocortical cells were obtained by fractionated trypsination of newborn rat adrenal glands and transfected with a plasmid containing the EJ T24 -Ha-ras oncogene. Isolation of adhesive cells led to a proliferative cell line with an overexpression of 21 kDa ras protein. These cells incubated with corticosterone or deoxycorticosterone as the precursor produced a high level of 18-hydroxycorticosterone and aldosterone as identified by gas chromatography- mass spectrometry. ACTH and angiotensin II increased the basal production of aldosterone nineteen-fold and six-fold respectively. Under ACTH stimulation the ratio between aldosterone and 18-hydroxycorticosterone production was 1:3. T…
Metabolism of Linoleic Acid or Mevalonate and 6-Pentyl-α-Pyrone Biosynthesis by Trichoderma Species
The understanding of the biosynthetic pathway of 6-pentyl-α-pyrone in Trichoderma species was achieved by using labelled linoleic acid or mevalonate as a tracer. Incubation of growing cultures of Trichoderma harzianum and T. viride with [U- 14 C]linoleic acid or [5- 14 C]sodium mevalonate revealed that both fungal strains were able to incorporate these labelled compounds (50 and 15%, respectively). Most intracellular radioactivity was found in the neutral lipid fraction. At the initial time of incubation, the radioactivity from [ 14 C]linoleic acid was incorporated into 6-pentyl-α-pyrone more rapidly than that from [ 14 C]mevalonate. No radioactivity incorporation was detected in 6-pentyl-…