Study on biotechnological production of biologically active substances from extracts of Japanese taxus trees.

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The aim of the study was biotechnological production of biologically active substances from extracts of Kyaraboku(Taxus cuspidata var.nana Rehd).1. Extracts in leaves and bark of kyaraboku.Two bisflavones (sciadopitysin and ginkgetin), two flavonols (isorhamnetin and quercetin), and two taxane-type of diterpenoids (taxinine and taxinine B) were isolated from the leaves of Kyaraboku. It was considered that taxinine could be converted to taxol mimic having anti-carcenogenic activity by enzymatic conversion with lipase.From the bark of Kyaraboku, taxol having strong anti-carcenogenic activity, was isolated in the yield of 0.001% to dry bark. The content of taxol in the bark was very small, however, it was considered that content of taxol could be increased by tissue culture of Kyaraboku.2.Pruduction of taxol mimic by enzymatic conversion of taxinine with lipase.Taxol mimic can be produced by reduction of enone carbonyl group in taxinine and followed by esterfication with lipase. Taxol mimic's side chain, phenylisoserine was synthesized from E-cinnamic acid through 8 steps in the total yield of 15%. Reduction of the carbonyl group with di-butylaluminium hydride are now being conducted.3. Production of taxol by tissue culture of Kyaraboku.Taxol was found to be produced in the calli induced from Kyaraboku stems and leaves. The amount of taxol in the calli was 1.1 times higher than that in intact plant. To increase the amount of taxol, the calli was eliciated with chito-oligosaccharides in various concentrations. The amount of taxol was rised up to 10 times higher than that of untreated calli.