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

这项研究的目的是从Kyaraboku(红豆杉变种nana Rehd)提取物中生物技术生产生物活性物质。从天南星的叶和树皮中分离得到两个双黄酮类化合物(接骨脂素和银杏素)、两个黄酮醇(异鼠李素和槲皮素)和两个紫杉烷型二萜类化合物(紫杉碱和紫杉碱B)。研究认为，紫杉醇可以通过脂肪酶催化转化为具有抗肿瘤活性的紫杉醇类似物。从嘉拉博库树皮中分离得到具有较强抗肿瘤活性的紫杉醇，产率为0.001%。紫杉醇在树皮中的含量很低，但人们认为通过组织培养可以提高紫杉醇的含量。2.紫杉醇的生产是通过脂肪酶催化紫杉碱的酶转化来生产的，紫杉醇的生产是通过还原紫杉醇中的烯酮羰基，然后用脂肪酶来酯化得到的。以E-肉桂酸为原料，经8步反应合成了紫杉醇类似物苯基异丝氨酸，总收率15%。目前正在用二丁基氢化铝还原羰基。紫杉醇的组织培养产生紫杉醇的研究结果表明，紫杉醇的产生是在其茎叶诱导的愈伤组织中进行的。愈伤组织中紫杉醇的含量是完整植株的1.1倍。为了增加紫杉醇的含量，用不同浓度的壳寡糖诱导愈伤组织。紫杉醇的含量比未经处理的愈伤组织高出10倍。

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.