黄连细胞二步法悬浮培养生产黄连生物碱类成分的探索

为提高用植物细胞培养生产次生代谢产物的单位产量，作者对黄连细胞采用二步法悬浮培养生产黄连生物碱进行了探索。结果显示：黄连细胞采用一步法悬浮培养６周，细胞干、鲜重产率分别为２０．９６ｇ／Ｌ和１７４．９２ｇ／Ｌ，增重约７００％；细胞中总生物碱含量为１４．７９ｍｇ／ｇ．ｃｅｌｄｗ，培养基中生物碱量为１２．９６ｍｇ／Ｌ，每升培养液共可收获生物碱３２３ｍｇ。采用先在生长培养基中培养３周，然后在合成培养基中培养３周的二步法培养，６周后收获细胞干、鲜重分别为１６．７２ｇ／Ｌ和１２７．４４ｇ／Ｌ，其生物量低于一步法，但总碱含量达３１．７６ｍｇ／ｇ．ｃｅｌｄｗ，培养基中生物碱量为２５．３１ｍｇ／Ｌ，每升培养液共可获生物碱５５６ｍｇ。二步法培养生物碱产率为一步法培养的１．７２倍。以上结果表明：二步培养法能够较好地解决黄连培养物生物量增长与次生代谢产物积累之间的矛盾，大大提高目的产物的产率，具有一定的实用价值。

Alkaloid Production of Cultured Coptis Cells by Two stage Suspension Culture

In this study the asexual cell line H292 induced and selected from Coptis gulinensis had rapid growth rate and could stably produce alkaloids. By the one stage method, after the cell suspensions were cultured on the same medium for six weeks, the increased dry and fresh weights of the cells were 20.96g/L and 174.92g/L respectively. The content of the total alkaloids in the cells was 14.79mg/g cell dw. Per litter li quid medium could provide 323 mg alkaloid. In contrast, the cells were cultured by two stage method. After having been cultured on the medium which contributed to the growth of the cells for three weeks, the cells were transferred to the medium which contributed to the production of the alkaloid and cultured for three weeks. Six weeks later, the dry and fresh weights of the cells were 16.72g/L and 127.44g/L, respectively. The biomass was lower than that by one stage method, but the content of the total alkaloids was as high as 31.76 mg/g cell dw, which was much more than that by one stage method. In addition, the content of the alkaloid in the medium was 25.31 mg/L. Per litter liquid medium could provide 556mg alkaloid. The total yield of alkaloid obtained by two stage method was 1.72 times than that by one stage method.