Lin Han-wei, Kwok Kian H, Doran Pauline M
School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney NSW 2052, Australia.
Biotechnol Prog. 2003 Sep-Oct;19(5):1417-26. doi: 10.1021/bp034036h.
Novel cross-species coculture systems using Linum flavum hairy roots and Podophyllum hexandrum cell suspensions were applied for in vitro production of podophyllotoxin. The hairy roots and suspensions were cocultured in Linsmaier and Skoog medium in dual shake flasks and dual bioreactors. In separate experiments, coniferin feeding was shown to be an effective strategy for increasing the accumulation of podophyllotoxin in P. hexandrum suspensions. Because roots of L. flavum are a natural source of coniferin, hairy roots of this species were used in coculture with P. hexandrum to provide an in situ supply of coniferin. Compared with P. hexandrum suspensions cultured alone in shake flasks or bioreactors, podophyllotoxin concentrations in cocultured P. hexandrum cells were increased by 240% and 72% in dual shake flask and dual bioreactor systems, respectively. The availability and stability of coniferin in the medium are the most likely factors limiting podophyllotoxin synthesis in coculture. Intensification of the coculture process is required to further improve total podophyllotoxin accumulation on a volumetric basis.
利用亚麻毛状根和桃儿七细胞悬浮液构建的新型跨物种共培养系统被用于鬼臼毒素的体外生产。毛状根和悬浮液在双摇瓶和双生物反应器中的Linsmaier和Skoog培养基中共培养。在单独的实验中，添加松柏苷被证明是一种有效提高桃儿七悬浮液中鬼臼毒素积累量的策略。由于亚麻根是松柏苷的天然来源，该物种的毛状根被用于与桃儿七共培养，以原位提供松柏苷。与在摇瓶或生物反应器中单独培养的桃儿七悬浮液相比，在双摇瓶和双生物反应器系统中，共培养的桃儿七细胞中鬼臼毒素浓度分别提高了240%和72%。培养基中松柏苷的可用性和稳定性最有可能是限制共培养中鬼臼毒素合成的因素。需要强化共培养过程以进一步提高单位体积内鬼臼毒素的总积累量。
