In vitro biosynthesis of isopentenylacetophenones in Eupatorium rugosum

A cell free homogenate of Eupatorium rugosum leaves was prepared and utilized to study the biosynthesis of dehydrotremetone (1). Homogenates of young leaves were most efficient in the formation of 1. The furan ring and its side chain were derived from an isoprenoid unit, which appeared to be isopentenyl pyrophosphate and not dimethylallyl pyrophosphate. Potential aromatic precursors such as 4-hydroxyacetophenone and 4-hydroxy-3[isopenten-(2)-yl]-acetophenone were poorly utilized. However, tremetone, especially in the presence of the coenzyme NADP, was very efficiently converted to 1. An apparent intermediate in the pathway leading to 1 was isolated and appeared similar to 4-hydroxy-3[isopenten-(2)-yl]-acetophenone. It is proposed that the aromatic moiety of 1 is derived from acetate via a polyketide intermediate, which undergoes isoprenylation by isopentenyl pyrophosphate, followed by aromatization and furan ring closure.     

In vivo synthesis of stigmasterol in Nicotiana tabacum

Six-day-old tobacco seedlings rapidly incorporated and metabolized exogenously supplied [4-¹⁴C]-sitosterol, but none of the radioactivity was recovered from stigmasterol. However, exogenously supplied [2-¹⁴C]-mevalonic acid was incorporated into both sitosterol and stigmasterol. Based on these results it is suggested that the biosynthetic pathway of stigmasterol is not via sitosterol but that both sterols have a common precursor.     

In vivo biosynthesis of tribromoheptene oxide in Bonnemaisonia nootkana

Experimental evidence is presented supporting the involvement of acetate and palmitate in the biosynthesis of tribromoheptene oxide in this red alga.     

Flavonoid constituents of Eupatorium odoratum

Isosakuranetin and a new chalcone, odoratin, have been isolated from the leaves of Eupatorium odoratum. The structure of odoratin has been shown to be 2′-hydroxy-4,4′,5′,6′-tetramethoxy chalcone.     

In vitro and in vivo biosynthesis of xanthophylls by the cyanobacterium Aphanocapsa

Of the six carotenoids identified in the cyanobacterium Aphanocapsa, β-carotene, zeaxanthin, echinenone and myxoxanthophyll are the major pigments, whilst β-cryptoxanthin and 3-hydroxy-4-keto-β-carotene are present only in trace amounts. With the exception of zeaxanthin, the other xanthophylls could be formed in vitro from [¹⁴C]phytoene in high yields, especially β-cryptoxanthin and 3-hydroxy-4-keto-β-carotene. In a time course experiment of xanthopyll biosynthesis the flow of radioactivity from [¹⁴C]phytoene was followed through the pools of phytofluene, lycopene, and β-carotene. The reaction sequence from phytoene to xanthophylls is sensitive in vitro to both difunone, an inhibitor of carotene desaturation, and CPTA, an inhibitor of cyclization.     

Two new flavones from Eupatorium coelestinum

Besides coumarin, nobiletin, lucidin dimethyl ether and 5,6,7,3′,4′-pentamethoxyflavone, two new highly oxygenated flavones were isolated from Eupatorium coelestinum. Their structures were determined by spectroscopic methods and alkaline degradations as 5,6,7,8,3′,4′,5′ -heptamethoxyflavone and 5,6,7,8,5′-pentamethoxy-3′,4′-methylenedioxyflavone.     

Biosynthesis of cyanidin in cell cultures of Haplopappus gracilis

-Dihydroquercetin proved to be a good precursor for light-stimulated cyanidin biosynthesis in cell suspension cultures of Haplopappus gracilis. Wit     

Ant-repellent sesquiterpene lactones from Eupatorium quadrangularae

The chloroform extract from the leaves of Eupatorium quadrangularae has been systematically fractionated by following biological activity in a bioassay which measures repellency to the leafcutter ant Atta cephalotes (Formicidae, Attini). Several sesquiterpene lactones were isolated, two of which showed significant ant-repellency.     

Biosynthesis of oleanolic acid glycosides by subcellular fractions of Calendula officinalis seedlings

The synthesis of oleanolic acid 3β-d-glucuronoside from oleanolic acid and UDPGlcA has been demonstrated in cell-free preparations from C. officinalis seedlings. Moreover, the formation of more complex glycosides by successive additions of galactose and glucose to oleanolic acid glucuronoside was observed when cell-free preparations were incubated with UDPGal or UDPGlc. The consecutive steps of oleanolic acid glycosylation are localized in three different cellular compartments. The biosynthesis of the 3-glucuronoside takes place in the microsomes, the elongation of the sugar chain at C-3 of the aglycone proceeds in heavy membrane structures which are probably fragments of the Golgi complex while a cytosol enzyme(s) is involved in glucosylation of the C-17 carboxyl group of oleanolic acid.     

Seven guaianolides from Eupatorium chinense

Two allyl hydroperoxy guaianolide sesquiterpene lactones (peroxyeupahakonin-A and -B) and five new guaianolides (eupahakonin-A and -B, eupahakonenin-A and -B, and eupahakonesin) were isolated from E. chinense and characterized. The allyl hydroperoxy sesquiterpene lactones were characterized by spectral and chemical methods. They were prepared chemically by photosensitized oxygenation of eupahakonin-A.     

Biosynthesis of limonoids in Citrus

Analyses of citrus leaves and fruits, and radioisotope tracer work showed that the leaves could synthesize limonoids and that the trees could translocate limonoids from leaves to the fruit. These results suggest that limonoids in citrus fruit tissues are synthesized in leaves and translocated to the fruit. No evidence was found to support the presence of limonoid biosynthetic systems in the fruit tissues.     

A glycosidic germacradienolide from Eupatorium altissimum

The isolation and structure determination of a glycosidic germacradienolide from Eupatorium altissimum are reported. Eupatorin and 5-hydroxy-3′,4′,6,7-tetramethoxyflavone were also found.     

Biosynthesis of citreomontanin in Penicillium pedemontanum

The ¹³C NMR spectrum of citreomontanin has been fully assigned by analysis in the presence of a shift reagent. The intramolecular distribution patt     

Biosynthesis of withanolides in Acnistus breviflorus

Administration of [2?¹⁴C]mevalonolactone to excised leaves of Acnistus breviflorus produced labelled withaferin A and jaborosalactone A. The former was degraded leading to the isolation of glyceric acid from C-25–C-27 of the withanolide. These carbons represented only 2 % of the total radioactivity of withaferin A. The relative radioactivity of these carbons indicated that C-26 is directly derived from C-2 of mevalonolactone suggesting that the 25-pro-R-methyl group of cholesterol or any other sterol intermediate had been oxidized to form the lactone ring of the withanolide. The total radioactivity value found for C-25–C-27 was much lower than the expected 20 % of the total value for the withanolide indicating that the side chain of the sterol precursor had been partially cleaved during the biosynthetic process.     

In vivo characterization of catechol ring-cleavage in cell cultures of Glycine max

No evidence could be obtained for the involvement of dioxygenases in the generation of ¹⁴CO₂ from catechol-[¹⁴C] observed in soybean cell cultures. Culture filtrates incubated with catechol and H₂O₂ were, however, able to catalyze the ring-cleavage, as was horseradish peroxidase.     

Biosynthesis of limonoids in Citrus seedlings

Radioactive tracer work showed that nomilinoate A-ring lactone was the predominant, if not the only, limonoid biosynthesized and accumulated in seedlings of lemon, Valencia orange, grapefruit and tangerine. Lemon seedlings were excellent tools for biosynthetic preparation of [¹⁴C]nomilin.     

Formation of natural thiophene derivatives from acetylenes by Tagetes patula

Six tritium-labelled thiophene derivatives were synthesized and their emulsions were applied to whole plants of Tagetes patula. From the root extra     

Biosynthesis of cardenolides in Digitalis lanata

[8-³H]-Cholesterol was synthesized. A doubly labelled sample of [8-³H, 4-¹⁴C]-cholesterol was administered to Digitalis lanata plants and the cardenolides were isolated. Biosynthesized digitoxigenin and digoxigenin retained all the tritium. Barring the migration of the tritium in biosynthesis the results are interpreted as indicative that neither intermediates with δ7, δ⁷ or δ⁸⁸⁽¹⁴⁾ are participating in the elaboration of cardenolides.     

Biosynthesis of thalicarpine in Thalictrum minus

Feeding experiments with ¹⁴C-labelled reticuline, protosinomenine, orientaline, their N-nor-analogues and ³H-labelled isoboldine have shown reticuline and isoboldine to be the most efficient precursors of thalicarpine in Thalictrum minus. A biosynthetic pathway for thalicarpine with reticuline and isoboldine at the benzylisoquinoline and aporphine stages respectively has been suggested. Support for this proposal has been provided by the demonstration by radioisotopic dilution that reticuline and isoboldine are minor constituents of the plant.