Olive (Olea europaea) is a well-known evergreen tree native to the Mediterranean coast, whose fruits and oil have been used for foods and cooking. Olive contains large quantity of triterpene acids including oleanolic acid (6) as a major one.1,2) Recently, much attention has been paid for triterpene acids from pharmaceutical viewpoints because of their anti-HIV, 3) anti-inflammatory, 4) anti-tumor-promoting 5) activities and antagonist activity for an endothelin receptor, 6) etc. Wax of olive fruits contains predominantly oleanolic acid, 1) whereas that of the leaves contains a mixture of oleanolic acid and betulinic acid (16) in a ratio of 7 : 2.2) In addition to these triterpene acids, triterpene-diols, such as erythrodiol (8: oleanane type) and uvaol (15: ursane type) are also contained, even though in less quantity than the corresponding acids. 2)Successful production of triterpene acids by plant cell cultures has been documented. Production of bryonolic acid (17), an anti-allergic triterpene acid, by cultured cells of several cucurbitaceous plants 7) is a good example. Cell cultures of plants, however, do not always produce the same secondary metabolites as those in the mother plants. Some callus cultures produce no trace of the metabolites found in the mother plants but produce those metabolites that are not produced by the mother plants. This situation is exemplified by Glycyrrhiza glabra cell cultures, which did not produce glycyrrhizin (18), a major triterpene saponin of the mother plant, but produced betulinic acid (16) 8) and soyasaponin I (19).9) It is quite interesting to know what kinds and how much of triterpenes cultured cells produce. If they produce any particular ones in large quantity, such production system would benefit the supply of triterpene acids for clinical use, since purification and isolation of triterpenes from complex mixtures are generally laborious. Thus we induced cell cultures of O. europaea and investigated the production of triterpene metabolites.Triterpenes are biosynthesized from a common precursor, (3S)-oxidosqualene (1). Over one hundred of diverse triterpene skeletons reported from nature 10) are constructed at the cyclization step of oxidosqualene catalyzed by oxidosqualene cyclase (OSC). Thus, the studies on their product specificity, in particular, would lead to better understanding of the structural diversity of natural triterpenes. As a continuation of our studies on OSCs 11) in this direction, cDNA clonig of OSCs from Olive cell cultures was also conducted.In this paper, we report induction of cell cultures, isolation Olive (Olea europaea) contains large quantity of triterpene acids including oleanolic acid (6) as a major one. Varieties of biological activities exhibited by triterpene acids attracted our attentions, especially from pharmaceutical viewpoints. Cell culture of olive plant was induced and its triterpene constituents were studied. From the cell suspension cultures, six ursane type triterpene acids; ursolic acid (9), pomolic acid (10), rotundic acid (11)...

Five phenylethanoid glycosides (acteoside, isoacteoside, beta-oxoacteoside, beta-hydroxyacteoside, and salidroside) were isolated from a cell suspension culture of Olea europaea. We examined the biosynthesis of acteoside in olive cell cultures by using feeding experiments with stable isotope labeled precursors. The hydroxytyrosol moiety of acteoside is biosynthesized from tyrosine through dopamine, whereas the caffeoyl moiety of acteoside is biosynthesized from phenylalanine via a cinnamate pathway. Dopamine is incorporated into acteoside through oxidation to the corresponding aldehyde, reduction to the alcohol, and then beta-glycosylation.

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Ganciclovir, foscarnet, vidarabine and ribavirin, which are used to treat viral infections in humans, inhibited the proliferation of a baculovirus (Bombyx mori nucleopolyhedrovirus) in BmN4 cells, a cultured silkworm cell line. These antiviral agents inhibited the proliferation of baculovirus in silkworm body fluid and had therapeutic effects. Using the silkworm infection model, the antiviral activity of Kampo medicines was screened and it was found that cinnamon bark, a component of the traditional Japanese medicine Mao-to, had a therapeutic effect. Based on the therapeutic activity, the antiviral substance was purified. Nuclear magnetic resonance analysis of the purified fraction revealed that the antiviral activity was due to cinnzeylanine, which has previously been isolated from Cinnamomum zeylanicum. Cinnzeylanine inhibits the proliferation of herpes simplex virus type 1 in Vero cells. These results suggest that the silkworm-baculovirus infection model is useful for screening antiviral agents that are effective for treating humans infected with DNA viruses.

Two high saponin-producing cell lines were selected from the original callus induced in 1967 and maintained in good state. For saponin production and growth in two cell lines, the comparison of static and suspension cultures, static cultures in the dark or in the light and rotary or reciprocal shaking cultures were made in detail. It was shown that the growth in rotary shaking cultures were 1.8 times higher than in reciprocal cultures, while the saponin production was almost the same, and the most effective hormonal condition was the combination of indole-3-butyric acid with kinetin. The saponin content per fresh weight was almost the same as that of the crude drug, whereas that per dry weight was 4 times higher.